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In the cytoplasm of virtually all clear-cell renal cell carcinoma (ccRCC), speckle-type POZ protein (SPOP) is overexpressed and misallocated, which may induce proliferation and promote kidney tumorigenesis. In normal cells, however, SPOP is located in the nucleus and induces apoptosis. Here we show that a structure-based design and subsequent hit optimization yield small molecules that can inhibit the SPOP-substrate protein interaction and can suppress oncogenic SPOP-signaling pathways. These inhibitors kill human ccRCC cells that are dependent on oncogenic cytoplasmic SPOP. Notably, these inhibitors minimally affect the viability of other cells in which SPOP is not accumulated in the cytoplasm. Our findings validate the SPOP-substrate protein interaction as an attractive target specific to ccRCC that may yield novel drug discovery efforts., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Objective: To investigate the treatment of surgical site infection(SSI) after spine surgery., Methods: Sixty-seven patients (aged 20-77 years with mean age of 51 years) with etiologically-confirmed surgical site infection after spine surgery in Peking University Third Hospital between July 2004 and December 2012 were retrospectively analyzed. There were 39 male and 28 female patients; 47 lumbar infections, 4 thoracic infections, 7 lower cervical infections and 8 upper cervical infections; 64 early infections and 3 delayed infections; 23 superficial infections and 44 deep infections; 47 monomicrobial infections and 20 polymicrobial infections. Ninety-six strains of bacteria were identified from the bacterial culture of 67 patients. Sixty strains were gram-positive pathogenic bacteria (62.5%), and the top three species were Staphylococcus aureus, S. epidermidis and Enterococcus faecalis. The remaining 36 strains were gram-negative pathogenic bacteria (37.5%), and the 3 species most predominant were Escherichia coli, Enterobacter cloacae, and Acinetobacter baumannii. All the patients with SSI were administered antibiotics. Debridement and irrigation-suction was performed if little symptomatic improvement was achieved in two-to-three days of antibiotics treatment, patients underwent 1-5 times of debridements (mean 1.5 times)., Results: One patient was dead of MRSA septicemia, whom manifested as high fever, alalia and incision swelling when the infection occurred. The patient underwent polymicrobial of pulmonary infection and urinary tract infection during the period of hospitalization, and finally died of multiple organ failure. Sixty-six cases had wound healed, and they were followed up for 25-117 months (average 70 months), no recurrence of infection was found at last follow-up. Among the 65 cases of internal fixation, 56 cases reserved the implants, while implants were removed in other 9 cases for controlling infection., Conclusion: Reasonable antibiotics and irrigation-suction are effective methods for managing surgical site infections after spine surgery and prevent removal of implants.

Objective: To analyze the etiology of instrumentation failure after corrective surgery for thoracolumbar focal kyphosis, and make suggestion for treatment., Methods: Retrospective study for 8 patients with thoracolumbar focal kyphosis who underwent surgical treatment and suffered instrumentation failure from June 2005 to December 2011 was made. The surgical procedures included pedicle subtraction osteotomy (PSO), anterior opening-posterior closing osteotomy and correction (AOPC), and posterior vertebral column resection (VCR). The reasons of instrumentation failure were analyzed and revision surgeries were performed., Results: The incidence of instrumentation failure was 6.3%. The average occurrence time was 22.5 months after surgery. Except one had failure in 3 months after surgery, all cases happened after 1 year. In this series, there were 5 cases with post-tuberculosis, 2 cases with post-traumatic kyphosis and 1 case with congenital kyphosis. For the surgical procedure, 7 cases underwent VCR and 1 case AOPC. After the instrumentation failure, all cases had back pain, and 3 of them had combined neurological symptoms. The reasons or risk factors of instrumentation failure included non-fusion of bone graft, VCR procedure, sink of the titanium mesh, insufficiency of anchor sites, and more severe kyphosis. All the 8 cases were treated with revision surgery and got good results., Conclusions: The instrumentation failure of thoracolumbar focal kyphosis is relatively late occurred, and can develop with various reasons. Positive revision surgery is suggested for the instrumentation failure, and good results can be expected.

A growing number of molecular epidemiological studies have been conducted to evaluate the association between human papillomavirus (HPV) infection and the malignancy of sinonasal inverted papilloma (SNIP). However, the results remain inconclusive. Here, a meta-analysis was conducted to quantitatively assess this association. Case-control studies investigating SNIP tissues for presence of HPV DNA were identified. The odds ratios (ORs) and 95% confidence intervals (CIs) were calculated by the Mantel-Haenszel method. An assessment of publication bias and sensitivity analysis were also performed. We calculated a pooled OR of 2.16 (95% CI=1.46-3.21, P<0.001) without statistically significant heterogeneity or publication bias. Stratification by HPV type showed a stronger association for patients with high-risk HPV (hrHPV) types, HPV-16, HPV-18, and HPV-16/18 infection (OR=8.8 [95% CI: 4.73-16.38], 8.04 [95% CI: 3.34-19.39], 18.57 [95% CI: 4.56-75.70], and 26.24 [4.35-158.47], respectively). When only using PCR studies, pooled ORs for patients with hrHPV, HPV-16, and HPV18 infection still reached statistical significance. However, Egger's test reflected significant publication bias in the HPV-16 sub-analysis (P=0.06), and the adjusted OR was no longer statistically significant (OR=1.65, 95%CI: 0.58-4.63). These results suggest that HPV infection, especially hrHPV (HPV-18), is significantly associated with malignant SNIP., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Objective: Extensive deep partial-thickness burns still seriously challenge the surgeon's abilities. This study aimed to assess the impact of early dermabrasion combined with porcine acellular dermal matrix (ADM) in extensive deep dermal burns., Methods: From September 2009 to September 2013, a total of 60 adult patients sustained greater than 50% total body surface area (TBSA) burn by hot water or gas explosion were divided into three groups based on dermabrasion: group A (early dermabrasion and porcine ADM), group B (early dermabrasion and nano-silver dressings), and group C (conservative group). The wound healing time and length of hospital stay were analyzed. Scar assessment was performed at 3 and 12 months after the injury with a modified Vancouver Scar Scale linked with TBSA (mVSS-TBSA)., Results: No significant difference was found in mean burn size, burn depth, age, male-to-female ratio, or incidence of inhalation injury between the patients in the three groups (p>0.05). Compared with groups B and C, the patients that received early dermabrasion combined with porcine ADM had a shorter wound healing time (p<0.01). The burn patients treated with early dermabrasion and porcine ADM coverage had a mean length of hospital stay of 28.3 days (±7.2), which was significantly shorter than that of groups B and C (p<0.05-0.01). The mVSS-TBSA of patients in group A was significantly improved in comparison with groups B and C at 3 and 12 months after the injury. There was no significant difference in the mortality rate between the three groups (p>0.05)., Conclusion: Early dermabrasion combined with porcine ADM coverage facilitates wound healing, reduces the length of hospital stay, and improves esthetic and functional results in extensive deep dermal burns with burn size over 50% TBSA., (Copyright © 2015 Elsevier Ltd and ISBI. All rights reserved.)

In this study, we determine the relationship between the expression of major histocompatibility complex class I chain-related gene A (MICA) in gastric cancer tumors after D2 gastrectomy and the clinical outcome of a CIK-containing adjuvant therapy. Ninety-five consecutive patients with gastric cancer after D2 gastrectomy who received adjuvant chemotherapy combined with CIK cell therapy were enrolled. The MICA expression of their tumors was determined by immunohistochemistry (IHC). High expression of MICA protein was documented by IHC in 38 of 95 tumor samples (40.0 %). The MICA status was significantly associated with the age and stage, p = 0.008 and 0.023, respectively. Analysis of NKG2D on in vitro expanded CIK cells showed that the percentages of NKG2D+ in CD3+/CD56+, CD3-/CD56+, and CD3+/CD8+ cells populations were 97.2 ± 1.4, 97.9 ± 1.8, and 95.6 ± 2.1 %, respectively. For patient with high MICA-expressing tumors, the median DFS and OS were longer than for the patients with tumors with low expression of MICA; 46.0 versus 41.0 months (p = 0.027), and 48.0 versus 42.0 months (p = 0.031), respectively. In a multivariate analysis, stage and MICA expression were independent prognostic factors for DFS and OS. Our findings show that adjuvant chemotherapy plus CIK therapy treatment is a promising modality for treating gastric cancer patients after D2 gastrectomy. Especially, those who have tumors with high expression of MICA were more likely to benefit from such a treatment strategy. Subsequent studies in clinical trial cohorts will be required to confirm the clinical utility of these markers.

The safety and efficacy of S-1 plus cisplatin in Chinese advanced gastric cancer patients in first line setting is unknown. In this pilot study, patients with advanced gastric or gastro-esophageal junction adenocarcinoma were enrolled and randomly assigned in a 1:1 ratio to receive S-1 plus cisplatin (CS group) or 5-FU plus cisplatin (CF group). The primary endpoint was time to progression (TTP). Secondary end points included overall survival (OS) and safety. This study was registered on ClinicalTrials. Gov, number NCT01198392. A total of 236 patients were enrolled. Median TTP was 5.51 months in CS group compared with 4.62 months in CF group [hazard ratio (HR) 1.028, 95% confidential interval (CI) 0.758-1.394, p = 0.859]. Median OS was 10.00 months and 10.46 months in CS and CF groups (HR 1.046, 95%CI 0.709-1.543, p = 0.820), respectively. The most common adverse events in both groups were anemia, leukopenia, neutropenia, nausea, thrombocytopenia, vomiting, anorexia and diarrhea. We find that S-1 plus cisplatin is an effective and tolerable option for advanced gastric or gastro-esophageal junction adenocarcinoma patients in China.

We aim to evaluate the efficacy and safety of paclitaxel-based doublet intravenous chemotherapy (IVC) with and without intraperitoneal chemotherapy (IPC) as the first-line treatment in advanced gastric cancer (AGC) with peritoneal metastasis (PM). 173 AGC patients with peritoneal metastasis were enrolled. All patients received paclitaxel-based doublet systemic chemotherapy Among them, 117 patients received IVC+IPC and 56 patients received IVC alone. The median OS of patients in the IP+ group was longer than the IP- group, however, there was no statistical difference between the two groups (11.1 months vs. 10.1 months, P = 0.072). In the multivariate analysis, the ECOG PS and IVC±IPC were independent prognostic factors for PFS and OS. There were no significant differences in the incidence of grade 3 and 4 toxicity between the IP+(DDP), IP+(FUDR) and IP- groups. Paclitaxel-based doublet regimens combining with IPC is effective, feasible and tolerated in AGC patients with PM.

To determine the relationship between the expression of phosphatase and tensin homologue (PTEN) and epidermal growth factor receptor (EGFR) in metastatic colorectal cancer (mCRC) and the clinical outcome of cetuximab-containing chemotherapy. A total of 158 consecutive mCRC patients with wild-type KRAS status who received chemotherapy with or without cetuximab, and for whom tumor tissue was available, were enrolled. The EGFR and PTEN expression was determined by immunohistochemistry (IHC). A total of 158 mCRC patients with wild-type KRAS status were enrolled in the study; 51 patients received chemotherapy combined with cetuximab, 107 patients received chemotherapy alone. Patients who received chemotherapy combined with cetuximab had longer overall survival (OS) compared with patients who received chemotherapy alone. High EGFR expression was detected in 60 patients (38.0%), while normal PTEN expression was detected in 60 patients (59.5%). The PTEN status was significantly related with the histological grade. For patients who received chemotherapy combined with cetuximab the median OS of patients with high-expression of EGFR was longer than the OS of patients with low EGRF expression; 25.0 versus 19.0 months, P = 0.002. For patient with normal PTEN the median OS were longer than the median OS for patients with loss of PTEN; 24.0 versus 19.0 months, P = 0.026. The overall response rate (ORR) had a borderline association with EGFR and PTEN expression (P = 0.055 and 0.048, respectively). In a multivariate analysis, ECOG PS, EGFR status, chemotherapy ± cetuximab, and the interaction of EGFR or PTEN and chemotherapy ± cetuximab were independent prognostic factors for OS. Our findings show that chemotherapy combined with cetuximab demonstrated encouraging antitumor activity for mCRC patients with wild-type KRAS status. Especially, those who have high EGFR expression or normal PTEN expression were more likely to benefit from such a treatment strategy. Subsequent studies in clinical trial cohorts will be required to confirm the clinical utility of these markers.

Cytokine-induced killer (CIK) cells have shown promising activity against gastric cancer in vitro and in vivo. Previous studies showed that cell signaling through MHC I-related Chain A (MICA)-Natural killer group 2, member D (NKG2D) results in CIK cell activation leading to cytolytic activities against tumor cells. In this study, we investigate the MICA status in patients with gastric carcinoma, and determine the potential relationship between MICA and clinical outcome of a CIK containing therapy. Two hundred and forty-three patients with gastric cancer who had received curative D2 gastrectomy were enrolled. The MICA expression of their tumors was determined by immunohistochemistry (IHC). Disease-free survival (DFS) and overall survival (OS) were evaluated. One hundred and forty-eight patients received adjuvant chemotherapy alone, and 95 patients received adjuvant chemotherapy combined with autologous CIK cell therapy. Patients who received adjuvant chemotherapy plus CIK had significantly longer DFS, 42.0 months vs. 32.0 months (P = 0.012), and OS, 45.0 months vs. 42.0 months (P = 0.039), by log-rank test. MICA high-expression, IHC scores of 5-7, was found in tumors from 89 of 243 patients (36.6%). The MICA expression was significantly correlated with the stage (P = 0.007) and there was a borderline association with histological grade (P = 0.054). In the adjuvant chemotherapy plus CIK group (n = 95), patients with high MICA expression had longer DFS, 46.0 months vs. 41.0 months (P = 0.027), and OS, 48.0 months vs. 42.0 months (P = 0.031). In the adjuvant chemotherapy alone group (n = 148), the median DFS and OS had no significant correlation with the MICA status. In a multivariate analysis stage, CIK therapy, and the interaction of MICA status and CIK therapy were independent prognostic factors for DFS and OS. Our study indicated that adjuvant chemotherapy plus CIK immunotherapy is a promising modality for treating gastric cancer patients after D2 gastrectomy. MICA status was associated with the outcome measures in CIK therapy, validation in prospective clinical trials is required to assess the value of this biomarker in the clinical decision-making process.

A polyoxovanadium borate (H2 dap)6H[V12B16O54(OH)4] · 12H2O (dap=1,2-diaminopropane) with novel structure was hydrothermally synthesized and characterized by the single crystal X-ray diffraction. It crystallizes in triclinic system with space group Piand unit cell parameters a=19.027(4), b=16.142(3) Å, c=26.679(5) Å, α=90°, β=101.06(3)°, γ=90°, V=8042(3) Å3, Z=4, Dc=1.962 g · cm(-3), μ=1.456 mm(-1), F(000)=4776, the final R1=0.0626, wR2=0.1927, S=1.003,for 7635 observed reflections with I>2σ(I). It is showed that the compound 1 is composed of V12B16 clusters unit and dap which is as a counter ion, and a two-dimensional layered structure is obtained by the effect of hydrogen bonding between the cluster units and dap, and between the layers via strong hydrogen bonds to form a three-dimensional supramolecular structure. The compound 1 were also characterized by IR, two-dimensional infrared (2D IR) correlation spectroscopy with magnetic and thermal perturbation, UV/Vis DRS spectra. The relationship between the structure and spectroscopy properties was discussed. The IR spectrum showed that the antisymmetric stretching vibration absorption peak νas (V-Oμ) and symmetric stretching vibration absorption peaks νs (V-Oμ) appeared at 775 and 683 cm(-1) respectively, whereas the vibration absorption peak in ν(B-O) of BO3 and the vibration absorption peak in ν(B-O) of BO4 appeared at 1350 and 1050 cm(-1) respectively. The response of the stretching vibrations of B-O and V-O was detected in the 2D IR correlation spectra with magentic perturbation. In addition, the response of the stretching vibrations of B-OH, B-O and V-O-V was detected in the 2D IR correlation spectra with thermal perturbation.

Background and Aims: It remains a challenge to prevent the local recurrence or distant metastasis of gastric carcinoma after D2 gastrectomy. Cytokine-induced killer (CIK) cells have shown promising activity against solid tumors in vitro and in vivo. We investigated the effect of adjuvant chemotherapy combined with autologous CIK therapy after D2 gastrectomy compared with adjuvant chemotherapy alone after D2 gastrectomy in patients with stage II-III gastric cancer., Methods: From January 2009 to December 2011, 226 patients with stage II-III gastric cancer who had had curative D2 gastrectomy were enrolled. Eighty-nine patients (CIK group) received adjuvant chemotherapy combining with autologous CIK therapy and 137 patients (control group) received adjuvant chemotherapy alone. Disease-free survival (DFS) and overall survival (OS) were evaluated., Results: Patients in the CIK group had longer DFS and OS than patients in the control group (DFS 41.0 months vs. 32.0 months, OS 45.0 months vs. 44.0 months, by log-rank test P = 0.006 and P = 0.028, respectively). In subgroup analysis, no significant differences in DFS and OS were observed between the two groups for the patients with stages II disease. Patients with stage III disease in the CIK group had longer median DFS and OS than patients in the control group (P = 0.031 and P = 0.038, respectively). In multivariate analysis, the stage and the interaction of stage and CIK therapy were independent prognostic factors for DFS and OS., Conclusions: The data suggest that adjuvant chemotherapy combined with autologous CIK therapy can improve prognosis for gastric carcinoma patients after D2 gastrectomy, especially for the patients with stage III disease.

Aims and Background: Peritoneal metastasis (PM) in patients with advanced gastric cancer (AGC) is a poor prognostic indicator. The aim of this study was to compare the response of AGC patients with PM to paclitaxel-based systemic multidrug chemotherapy with and without additional intraperitoneal (IP) chemotherapy through retrospective analysis., Methods and Study Design: Two hundred and sixty-three AGC patients with PM were enrolled. Eighty-two patients received systemic paclitaxel/oxaliplatin and leucovorin/5-fluorouracil (POF) and 181 patients received 2-drug systemic therapies, PO (paclitaxel + oxaliplatin) or PF (paclitaxel + 5-fluorouracil + leucovorin), and IP infusion of a third drug., Results: Patients who received the POF regimen had longer progression-free survival (PFS) and overall survival (OS) than patients who received PO/PF + IP therapy (P = 0.026 and P = 0.046), respectively. In subgroup analysis, no significant differences in PFS and OS were observed between the POF regimen and PF/PO + IP regimens in patients with isolated peritoneal metastatic disease. Patients with multiorgan metastatic disease receiving POF had better PFS and better OS than patients receiving PO/PF + IP chemotherapy (P = 0.005 and P = 0.036, respectively). In multivariate analysis, ECOG performance status and the interaction between different therapeutic strategies and multiorgan metastasis were independent prognostic factors for survival. Leukopenia, fatigue and peripheral neuropathy were higher on the triplet regimen than the doublet regimens., Conclusions: Paclitaxel-based doublet therapy combined with IP chemotherapy had more manageable toxicity and equal efficiency compared to triplet therapy for AGC patients with isolated PM. The POF regimen may be a good choice for AGC patients with multiorgan metastasis, especially those having a good performance status.

A phytochemical investigation on the aerial parts of a Tibetan medicine Meconopsis horridula, by solvent extraction, repeated chromatographies on silica gel, Sephadex LH-20, and preparative TLC techniques, led to the isolation of 9 compounds. By spectroscopic analysis and comparison of its 1H and 13C-NMR data with those in literatures, their structures were identified as oleracein E(1), N-( trans-p-coumaroyl) tyramine (2), chrysoeriol (3), apigenin (4), hydnocarpin (5), p-coumaric acid glucosyl ester (6), stigmast-5-ene-3beta-ylformate (7), 3beta-hydroxy-7alpha-ethoxy-24beta-ethylcholest-5-ene (8), and beta-sitosterol (9), respectively, among which compounds 6-8 were isolated from the genus for the first time,and 1,3 were isolated from the species for the first time. A MTT method was applied to evaluate the cytotoxic activity of compounds 14 against the human hepatocellular liver carcinoma cell line (HepG2), and compound 1 showed significant cytotoxicity against HepG2,with its inhibitory rate of 52.2% at 10 micromol x L(-1).

Litsea cubeba is one of aromatic medicinal plant belonging to family Lauraceae. The roots, stems and fruits of L. cubeba have been widely applied as folk medicines in some districts in China for relieving rheumatism and cold, regulating Qi (meridian) to alleviate pain. Previous studies revealed that this species contains major alkaloids, in specific aporphines, and minor flavonoids, lignans as well. Related pharmacological investigations demonstrated its activities and clinical applications on cardiovascular diseases, anti-cancer, against rheumatoid arthritis, relieving asthma and anti-allergic effects, as anti-oxidants, and so on. As an effort for further exploration of this bioactive ingredients and potential drug development, this paper summarizes most phytochemical and pharmacological results. Further, future prospects are also included.

Objective: To investigate the clinical significance of the expression of MHC class I chain-related gene A (MICA) in patients with advanced non-small cell lung cancer and explore the relationship between MICA expression and the efficacy of cytokine-induced killer cell (CIK) therapy for treating advanced non-small cell lung cancer., Methods: We obtained data on 222 patients with advanced non-small cell lung cancer, including data on MICA expression, age, gender, ECOG score, pathological type, stage, treatment history (including 38 patients who were given autologous CIK cell infusion), and overall survival (OS). MICA expression in lung cancer tissue was evaluated by immunohistochemical staining. Analyses of MICA expression, and CIK therapy association with survival outcomes were performed using Cox proportional models, Kaplan-Meier methods, and the log-rank test., Result: s MICA was expressed in both membrane and cytoplasm. MICA expression correlated with the stage of lung cancer, ECOG score, gender and age. Multivariate COX regression analysis showed that the expression of MICA was an independent prognostic factor of advanced non-small cell lung cancer (p = 0.002). In subgroup analysis, we divided the 222 patients into CIK and control groups. In the CIK group, the medium OS (mOS) of patients with a high expression of MICA was longer than in those with low expression of MICA (27 months vs. 13 months). In the control group, the mOS in patients with a high expression of MICA was shorter than in patients with low MICA expression (9 months vs. 18 months). COX regression analysis showed that the MICA expression affects the effect of CIK therapy (p<0.0001)., Conclusion: 1) The high expression of MICA is one of the indicators of a poor prognosis for advanced non-small cell lung cancer patients. 2) The high expression of MICA might be one of the predictive factors for successful CIK therapy.

A new triphenylamine-bridged fluoranthene derivative, 4-(7,10-diphenylfluoranthen-8-yl)-N-[4-(7,10-diphenylfluoranthen-8-yl)phenyl]-N-phenylaniline (BDPFPA), with a high glass transition temperature of 220 °C has been synthesized and characterized. BDPFPA is a highly fluorescent and versatile material that can be used as a nondoped green emitter and as a hole transporter. BDPFPA was used in a standard trilayer device as the emitting layer, which showed a low turn-on voltage (<3 V) and a high efficiency of 11.6 cd A(-1). The device also shows little efficiency roll-off at high brightness. For example, the efficiency can still be maintained at 11.4 cd  A(-1) (5.4 lm  W(-1)) at a brightness of 10,000 cd  m(-2). These results are among the best reported for nondoped fluorescent green organic light-emitting diodes. A simple bilayer device, in which BDPFPA serves as a hole-transporting layer, has a maximum power efficiency of 3.3 lm  W(-1) and the performance is nearly 40 % higher than that of an N,N'-bis(1-naphthyl)-N,N'- diphenyl-1,1'-biphenyl-4,4'-diamine (NPB)-based standard device., (Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Purpose: To report the results of a safety analysis from a phase II trial comparing administration of weekly paclitaxel plus S-1 (TS) versus paclitaxel plus 5-fluorouracil (5-FU)/calcium folinate (LV) (TLF) as first-line therapy for advanced gastric cancer., Methods: Patients (n = 240) with previously untreated advanced gastric cancer were randomly assigned to receive either TS or TLF in a 28-day cycle for six cycles., Results: The clinical features of both sets of patients were similar, with the exception of the incidence of prior chemotherapy (P>0·05). Most treatment-related adverse events occurred at similar rates in both treatment arms. However, patients receiving TS experienced an increase in all-grade especially grade 3/4 neutropenia, with an incidence of 43·7% in the TS arm and 16·3% in the TLF arm, respectively (P<0·05). Other severe adverse events were infrequent and not significantly different between the groups., Conclusion: The safety and tolerance of weekly paclitaxel plus S-1 or 5-FU/LV is well in untreated advanced gastric cancer patients.

Objective: To evaluate the efficacy and safety of weekly paclitaxel combined with S-1 or fluorouracil in the first line treatment of advanced gastric carcinoma., Methods: Two hundred and forty patients with untreated advanced gastric carcinoma were randomized into two arms, patients in the experimental arm were given paclitaxel and S-1, while those in the control arm received paclitaxel and fluorouracil. The regimen of experimental arm was paclitaxel 60 mg/m(2) by intravenous infusion, day 1, 8, 15; S-1 80 - 120 mg/day given by oral administration, day 1 - 14. The regimen of control arm was fluorouracil 500 mg/m(2) by intravenous infusion continuously, day 1 - 5; CF 20 mg/m(2) by intravenous infusion, day 1 - 5. The regimens in both arms were repeated every 28 days. The efficacy and safety of both arms were assessed., Results: Two hundred and twenty-eight patients were analyzed in the full analysis set, and 192 patients were analyzed in per-protocol set (experimental arm 100 patients, control arm 92 patients). The overall response rates of experimental and control arms were 50.0% and 28.3% (P = 0.002), and the disease control rates were 82.0% and 70.7% (P = 0.064), respectively. The primary endpoints of experimental arm were non-inferior to that of the control arm. The secondary endpoint of experimental arm in terms of median progression free survival was significantly better than that of control arm (5 months versus 4 months, P = 0.006). The experimental arm had a higher incidence of grade III-IV bone marrow suppression than the control arm, but the incidence of fever in both arms was not significantly different., Conclusions: Oral administration of S-1 is an alternative option of venous infusional fluorouracil. Weekly paclitaxel combined with S-1 is a safe regimen and has a promising efficacy.

We investigated the importance of HMGN5, a nuclear protein that binds to nucleosomes, unfolds chromatin, and affects transcription, in the LNCaP prostate cancer cell line. We also examined the molecular mechanisms that promote apoptosis of LNCaP cells after infection with small interfering RNA (siRNA) targeting HMGN5 (siRNA-HMGN5). The androgen-dependent LNCaP human prostate cancer cells were infected with siRNA-HMGN5. Apoptosis was detected using the Annexin V-PE/7-AAD double staining and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assays. Mitochondrial membrane potential was measured by JC-1 staining. HMGN5 and GAPDH mRNA expression were determined using real-time PCR. Bcl-2 and other apoptosis-related protein levels were determined by Western blot analysis. Caspase activity was measured by cleavage of the caspase substrate. Infection with siRNA targeting HMGN5 efficiently and specifically reduced the HMGN5 expression in LNCaP cells. The downregulation of HMGN5 induced remarkable apoptosis of LNCaP cells and resulted in the reduction of mitochondrial membrane potential. The induction of cell apoptosis was accompanied by the upregulation of Bax, the Bax/Bcl-2 ratio and the activation of caspase3. The HMGN5-targeted siRNA was effective in downregulating the expression of HMGN5 in androgen-dependent prostate cancer cells and inducing cell apoptosis via the regulation of a caspase-related mitochondrial pathway and Bcl-2 family proteins. This study suggests that HMGN5 may be a potential molecular target with therapeutic relevance for the treatment of prostate cancer.

Objectives: To investigate the long-term surgical outcome of thoracic myelopathy caused by the ossification of the ligamentum flavum (OLF) and evaluate the related risk factors., Methods: Forty-four patients who underwent decompressive laminectomy with thoracic OLF between January 1990 and December 2005 and got more than 5 years follow-up were retrospectively reviewed. Among these 44 cases, there were 29 male and 15 female whose ages at operation were 52 years averagely (27-68 years). The 2-year follow-up results and long-term outcomes were classified according to the modified Epstein's standard, and then the rates of excellent or good (REG) were calculated. The correlation between the long-term REG and the patients' ages, durations of symptoms, decompressed levels, and dural leak were analyzed., Results: The mean follow-up period of these 44 cases was 8.5 years (5-19 years). The REG at 2 years after laminectomy was 77.3% (34/44), while the long-term REG was 65.9% (29/44). There was one case who had suffered from an acute spinal cord injury got a poor post-operative outcome. The other 43 cases had chronic durations, including 22 cases whose pre-operative durations of symptoms were less than 12 months and 21 cases whose durations were equal to or more than 12 months. And the long-term REG of these two groups were 77.3% (17/22) and 57.1% (12/21) respectively (P>0.05). The REG of those cases whose decompression levels were limited in T1-T9 was 78.9% (15/19), while that of those cases whose laminectomy was relevant to thoracolumbar segment (T10-L2) was 58.3% (14/24) (P>0.05). There were 7 cases who had excellent or good short-term results and poor long-term outcomes. The reasons of these changes included coexistence of lumbar spinal stenosis in three cases and the growth of the OLF at the adjacent levels in four cases., Conclusions: Although the short-term results of the decompressive surgery for thoracic OLF is good, the regular long-term follow-up is necessary because the symptoms may reoccur or deteriorate secondary to lumbar spinal stenosis or the growth of OLF at the adjacent levels near former decompressive levels; the duration of symptoms which is more than one year and the decompression levels that is involved to T10-L2 segments are possibly related to the poor long-term outcomes.

Background: Platelet aggregation mediated by inflammation played a critical role in the development of coronary heart diseases (CHD). Our previous clinical researches showed that Th17 cells and their characteristic cytokine IL-17A were associated with the plaque destabilization in patients with acute coronary syndrome (ACS). However, the potent effect of IL-17A on platelets-induced atherothrombosis remains unknown., Methods and Results: In this study, we detected the plasma IL-17A levels and platelet aggregation in patients with stable angina (SA), unstable angina (UA), acute myocardial infarction (AMI) and chest pain syndrome (CPS). In addition, the markers of platelet activation (CD62P/PAC-1) and the mitogen-activated protein kinases (MAPKs) pathway were detected in platelets from ACS patients. We found that plasma IL-17A levels and platelet aggregation in patients with ACS (UA and AMI) were significantly higher than patients with SA and CPS, and the plasma IL-17A levels were positively correlated with the platelet aggregation (R = 0.47, P<0.01). In addition, in patients with ACS, the platelet aggregation, CD62P/PAC-1 and the phosphorylation of ERK2 signaling pathway were obviously elevated in platelets pre-stimulated with IL-17A in vitro. Furthermore, the specific inhibitor of ERK2 could attenuate platelet aggregation and activation triggered by IL-17A., Conclusion: Our experiment firstly proved that IL-17A could promote platelet function in patients with ACS via activating platelets ERK2 signaling pathway and may provide a novel target for antiplatelet therapies in CHD.

Objective: To observe the results of posterior osteotomy and correction in the surgical treatment of old tuberculous kyphosis., Methods: From June 2004 to December 2008, 31 cases of old tuberculous kyphosis with posterior osteotomy and correction technique were treated. There were 12 cases of male and 19 cases of female. The average age was 33.4 years. Pedicle subtraction osteotomy or vertebral column resection were applied in surgery. The kyphosis angle, lumbar lordosis angle and sagittal balance condition of the spine were measured before and after surgery, as well as follow-up. The Frankel grading system for neurological function of lower extremities, the Oswestry disability index (ODI) for life quality, and patient satisfactory index (PSI) for satisfaction of surgery were applied before surgery and at follow-up., Results: The average kyphosis angle was 94° ± 27°, the average lumbar lordosis angle was 71° ± 20°, and the average sagittal C(7) plumb line was (-15 ± 44) mm away from the balance region before surgery. The average kyphosis angle decreased to 26° ± 11° in one week after surgery, with an improvement rate of 71.4%. The average follow-up time was 22.5 months. The average kyphosis angle was 28° ± 12° at the final follow-up, with an improvement rate of 70.0%. The average lumbar lordosis angle was 46° ± 11°, with an improvement rate of 35.1%. The postoperative kyphosis angle and lumbar lordosis angle were significantly different with that of pre-operation (for kyphosis angle: t = 16.3, P < 0.05; for lumbar lordosis angle: t = 8.1, P < 0.05). The average sagittal C(7) plumb line was (-4 ± 22) mm away from the balance region at the final follow-up, with an improvement rate of 73.4%. The Frankel grading were E in 13 cases, D in 13 cases, and C in 5 cases before surgery, and were E in 20 cases, D in 8 cases, and C in 3 cases at the final follow-up. The average ODI was 13 ± 12 before surgery, and was 7 ± 8 at the final follow-up, with an improvement rate of 45.2%. The PSI results showed a satisfied rate of 90.3%., Conclusion: Good results can be achieved by applying proper posterior osteotomy and correction technique according to the severity of old tuberculous kyphosis.

Background: Various surgical approaches have been successfully used in the treatment of thoracolumbar disc herniation (TLDH). Although the anterior transthoracic approach has a reputation for better visualization than the posterolateral and lateral approaches, it involves the manipulation of the thoracic and pulmonary structures. Thus, this approach is technically demanding and prone to compromising the respiratory system. An ideal approach would involve adequate visualization and be accomplished through the posterior midline approach that is familiar to spine surgeons. The objective of this retrospective preliminary clinical study was to introduce a new surgical procedure, circumspinal decompression through a single posterior incision, for the treatment of TLDH (T10/11-L1/2) and to evaluate the surgical outcome of this procedure by comparing it to the conventional anterior transthoracic approach., Methods: In this study, 15 patients (10 males, 5 females; mean age 51 years) with symptomatic TLDH underwent the circumspinal decompression through a single posterior incision procedure between January 2008 and December 2009. Altogether, 17 herniated discs were excised, with 2 discs at T10/11, 4 discs at T11/12, 5 discs at T12/L1 and 6 discs at L1/2. Of these patients, 13 were followed up with a mean follow-up period of 23.5 months. Clinical outcomes, including operative time, blood loss, perioperative complications, postoperative time of hospitalization, neurologic status improvement, back pain and correction of local kyphosis, were investigated by comparing these data with the results from patients who underwent the anterior transthoracic approach for TLDH during the same period. The patients' neurologic status was evaluated by a modified Japanese Orthopedic Association (JOA) scoring system of 11 points. Neurologic status improvement after the surgery was assessed by calculating the recovery rate, which was equal to the (postoperative JOA score-preoperative JOA score)/(11-preoperative JOA score)×100%. The rates of patients who improved at the final follow-up were also assessed., Results: The mean operative time was 183 minutes, the mean blood loss was 1067 ml, and the mean postoperative hospitalization time was 8.4 days. Three patients suffered perioperative complications, but none of these complications involved the respiratory system. Local kyphotic angles at the fusion levels were reduced. Of the 13 patients that were followed up, 12 improved at the final follow-up, with a mean recovery rate of 52.8%. Patients who underwent the circumspinal decompression procedure showed a higher percentage of improvement at the final follow-up, a higher degree of local kyphosis correction and a lower percentage of complications (especially respiratory complications) compared to patients who underwent the anterior transthoracic decompression procedure., Conclusions: The circumspinal decompression through a single posterior incision procedure is an effective and safe technique that is comparable to anterior tranthoracic approach for the surgical treatment of TLDH patients. It could be an attractive choice in certain circumstances.

Object: In this paper, the authors aimed to summarize the clinical characteristics of ossification of the ligamentum flavum (OLF) associated with dural ossification (DO) and to identify improved methods for preoperative diagnosis., Methods: Thirty-six patients who had undergone OLF surgery between February 2005 and September 2009 were included in this retrospective study. The patients were divided into 2 groups: one that included patients with intraoperative evidence of DO and a second group that included patients without DO. The clinical features of DO were summarized and the neurological status of the patients was evaluated pre- and postoperatively., Results: The incidence rate of DO associated with OLF was 39% (14/36). The sensitivity and specificity of the tram track sign were found to be 93% and 59%, respectively. Dural ossification was found among 86% of the patients with tuberous type Sato classification. The postoperative neurological status of patients was generally improved relative to that observed prior to surgery, although neurological recovery did not differ between the 2 groups. Cerebrospinal fluid leakage was the main complication, occurring predominantly in the patients with DO, and all leaks resolved in all patients after comprehensive treatments., Conclusions: The tram track sign and Sato classification were found to be useful for preoperative diagnosis of DO and for determining the surgical procedure to be performed. Dural ossification had no effect on postoperative neurological recovery.

Objective: To analyze the impact of thoracic and thoracolumbar angular kyphosis on pelvic shape and sagittal alignment., Methods: From May 2002 to June 2010, the sagittal spino-pelvic parameters were analyzed in lateral standing radiographs of 32 patients (mean age 29.6 years) with thoracolumbar angular kyphosis. The parameters included Cobb angle of kyphosis, lumbar lordosis (LL), pelvic incidence (PI), sacrum slope (SS), pelvic tilt (PT) and sagittal vertical axis (SVA). All pelvic parameters in the patients were compared with those reported in historical normal subjects. All patients were treated by using kyphotic correction and fusion. The preoperative and postoperative parameters were compared. The pelvic parameters were also compared between the patients with kyphotic apex located at T(1-8) and those located at T(9-12) and thoracolumbar junction. The linear regression analysis was used to investigate the independent factors of PI., Results: The mean kyphosis was 90.1° (31° - 138°). The mean age of kyphosis occurrence was 6.1 years. The mean PI, SS and PT were 34.8°, 35.8° and -0.7° respectively. The PI and PT were significantly smaller (P < 0.001) in the patients than those in normal subjects while the SS was similar. The kyphosis was improved to 27.9° post-operatively. There was no difference in PI values between pre-operation and postoperation (P > 0.05). The PI and SS in patients whose kyphosis located at thoracic spine (T(1-8)) were significantly higher than those at T(9)-L(2). Instead of patients' age and LL, the preoperative Cobb angle of kyphosis and the levels where kyphosis located were two independent impact factors of PI., Conclusions: The kyphosis occurred at childhood may influence pelvic shape and alignment significantly. The lower kyphotic apex located and the bigger kyphosis, the greater impact on the pelvic morphology. The surgery can improve the kyphosis, but can not change the sagittal pelvic morphology. Early treatment of thoracolumbar angular kyphosis is beneficial not only to reconstruction of spine alignment but also to the formation of sagittal pelvic morphology.

Objective: To Evaluate the incidence rates and extents of deglutition disorder in patients with laryngeal carcinoma after different types of supracricoid partial laryngectomy., Methods: Retrospective analysis of postoperative deglutition disorder in patients with laryngeal carcinoma after different types of supracricoid partial laryngectomy treated in our department from 2005 to 2009. The extents of postoperative deglutition disorder were evaluated using a homemade quantitative score table at 5-20 days postoperation., Results: The score of deglutition disorder was 2.71 ± 0.31 in the supracricoid partial laryngectomy-cricohyoidoepiglottopexy (SCPL-CHEP) operation group and 3.43 ± 0.64 in the supracricoid partial laryngectomy-cricohyoidopexy (SCPL-CHP) group, respectively. The deference was statistically significant between the two groups (P < 0.001). The coefficient between age and score of postoperative deglutition disorder was assessed by Pearson correlation analysis. The coefficient of correlation was 0.947 (P < 0.0001) in the SCPL-CHEP group and 0.907 (P < 0.0001) in the SCPL-CHP group. The incidence rate of deglutition disorder was 1/37 in the SCPL-CHEP group and 7/30 in the SCPL-CHP group, evaluated at 8 weeks postoperation (P = 0.012). The deference between the two groups was significant., Conclusions: The type of operation procedure is an important factor affecting the occurrence of postoperative deglutition disorder in the patients after supracricoid partial laryngectomy, more serious in the SCPL-CHP group. The severity of postoperative deglutition disorder is more serious along with the increase of patient's age. For the elderly (> 70 years of age) patients with laryngeal carcinoma, the choice of surgical procedure should be more cautious, especially with the SCPL-CHP operation.

Objective: To investigate the effects of interactions among environmental factors, bone morphogenetic protein-4 (BMP4) and transforming growth factor beta-3 (TGF-β(3)) polymorphisms on nonsyndromic cleft lip and cleft palate (NSCLP)., Methods: The data of environmental exposures were collected with questionnaires. Genotypes were determined with techniques of polymerase chain reaction-restriction fragment length polymorphism. Interactions between genes, environmental factors and NSCLP were analyzed using multifactor dimensionality reduction (MDR) method. The interactions were validated by logistic regression analysis., Results: There was no correlation between three single nucleotide polymorphism (SNP) associated with NSCLP. The developmental accident of NSCLP had higher risk in the interaction between BMP4 T538C, maternal passive smoking and infection in first trimester pregnancy, as well as in the interaction of six factors between TGF-β(3) G15572-, maternal passive smoking, infections, multivitamin supplement in the first trimester pregnancy, paternal smoking and high risk drinking before realizing pregnancy than in other interactions of environmental factors. The results could be validated by logistic regression analysis., Conclusions: The NSCLP is induced by the interactions between genes and environmental risk factors.

Objective: To evaluate the clinical outcomes of posterior surgical corrective methods for ankylosing spondylitic kyphosis., Methods: From June 2003 to June 2008, 21 cases of ankylosing spondylitic kyphosis received posterior surgical correction. There were 17 male and 4 female, and the average age was 39.5 years (range, 20 to 57 years). The total spine X-ray and CT were used to evaluate sagittal balance and thoracolumbar spine kyphosis angle, and chin brow-vertical angle was obtained from clinical lateral photograph. The surgical goal was to correct sagittal imbalance and chin brow-vertical angle. The simulated osteotomy was performed in computer before surgery to determine the correction methods. The surgical methods included: 16 cases of monosegmental closing osteotomy correction, 3 cases of anterior opening-posterior closing osteotomy correction, and 2 cases of combined pedicle subtraction osteotomy in thoracolumbar spine and Smith-Peterson osteotomy in lumbar spine. All patients were followed up after surgery, and the improvement of sagittal imbalance, chin brow-vertical angle and thoracolumbar spine kyphosis angle were assessed. The symptoms relief and satisfied rate were also evaluated., Results: The average operation time was 4.4 hours, and the average blood loss was 1770 ml. Before surgery, the average thoracolumbar kyphosis angle was 62.1°, the average anterior shift of C(7) plumb line was 172.9 mm, and the average chin brow-vertical angle was 34.9°. The average follow-up was 28.8 months after surgery. The average correction rate of thoracolumbar kyphosis angle was 60%, the average improvement rate of anterior shift of C(7) plumb line was 64%, and the average correction rate of chin brow-vertical angle was 98%. The improvement rate of back pain was 64% during follow-up. The total surgical satisfactory rate was 95%., Conclusion: Based on the simulated osteotomy in computer before surgery, according to the characteristics of ankylosing spondylitic kyphosis, different posterior osteotomy and correction methods can achieve good results.

A Cu(2+)-specific colorimetric sensor 1, which is stabilized by an intramolecular hydrogen bonding, was designed and developed. The color of 1 changes from purple to blue on addition of 1.0 muM Cu(2+) in aqueous buffer solution, which can be detected by the naked eye. The analytical detection limit for Cu(2+) by the naked eye is as low as 1.0 muM. The stoichiometry for 1 and Cu(2+) in complex is 2:1 in aqueous solution.

Objective: To compare the results of extended vertical partial laryngectomy (similar to modified supracricoid partial laryngectomy with cricohyoidoepiglottopexy) and cricohyoidoepiglottopexy in the treatment of laryngeal carcinoma., Methods: Retrospectively analyzed on the results and prognosis in patients underwent extended vertical partial laryngectomy and cricohyoidoepiglottopexy between 1998 and 2005. The operation was similar to extended vertical partial laryngectomy. The healthy vocal cord and ventricular band as well as about 1/3 to 2/3 laminas of thyroid cartilage were removed. The healthy cricoarytenoid joint was reserved. The vocal cord, ventricular band, fixed or limitation of motion arytenoid cartilage and 2/3 laminas of thyroid cartilage in ill side were removed. The posteroinferior border of laminas of thyroid cartilage in both sides were reserved. The cricoid was lifted and fixed with hyoid epiglottis directly. Extended vertical partial laryngectomy group consisted of 37 patients with glottic carcinoma (stage T2 16 cases, stage T3 21 cases) and cricohyoidoepiglottopexy group consisted of 34 patients with glottic carcinoma (stage T2 12 cases, stage T3 21 cases, stage T4 1 case)., Results: Kaplan-Meier analysis was performed to calculate the survival rates. The three-year cumulative survival rate was 91.7% in extended vertical partial laryngectomy group and 87.5% in cricohyoidoepiglottopexy group respectively. There was no significant difference between the two groups (P > 0.05). The five-year cumulative survival rate was 80.6% in extended vertical partial laryngectomy group and 81.3% in cricohyoidoepiglottopexy group respectively. There was also no significant difference between the two groups (P > 0.05). The decannulation rate was 100% (37/37) in extended vertical partial laryngectomy group and 94.1% (32/34) in cricohyoidoepiglottopexy group respectively. The decannulation time was (14.0 + or - 2.3) days in extended vertical partial laryngectomy group and (19.0 + or - 4.6) days in cricohyoidoepiglottopexy group respectively. The incidence of aspiration was 2.7% (1/37) in modified group and 23.5 (8/34) in cricohyoidoepiglottopexy group respectively evaluated at 8th weeks post-operatively. The evaluation of deglutition disorder was analyzed by Ridit analysis in both groups and the results showed that there was significant difference between the two groups (U = 7.341, P < 0.001). The symptom of aspiration in extended vertical partial laryngectomy group was significant less than in cricohyoidoepiglottopexy group., Conclusions: Although the survival rate was not different between the two groups. The preservation of laryngeal function in extended vertical partial laryngectomy group was significant better than in cricohyoidoepiglottopexy group and extended vertical partial laryngectomy.

Due to the insufficient supply of embryonated chicken eggs, the preparation of large quantities of inactivated influenza vaccines will require an alternative virus culture system after the emergence or reemergence of a pandemic influenza virus. The Vero cell is one of the ideal options since it was used for producing many kinds of human vaccines. However, most of the influenza viruses can not grow well in Vero cells. To develop a new influenza vaccine with Vero cells as a substrate, the virus needs to adapt to this cell substrate to maintain high growth characteristics. By serial passages in Vero cells, the B/Yunnan/2/2005va (B) strain was successfully adapted to Vero cells, with the hemagglutination titer (HAT) of the virus reaching 1:512. The high growth characteristic of this strain is stable up to 21 passages. The strain was identified by hemagglutination inhibition (HAI) test and sequencing respectively; the HA₁ gene sequence of the virus was cloned and analyzed. The screening and establishment of high growth B virus provides an important tool for influenza vaccine production in Vero cells.

By using 3-year field experimental results and related meteorological observation data, the dynamic characteristics of leaf area index (LAI) and the allocation characters of ecological resources for different yielding spring maize (Zea Mays L.) population in Huadian of Jilin Province were studied. The results showed that the dynamic characteristics of relative LAI, with the relative growth days of test population, relative effective accumulated temperature, relative sunshine hours and relative rainfall as independent variables, fitted rational formula y = (a + bx) /(1 + cx + dx2), and the regression equation of maize yield with the ratios of growth days before and after silking (x1), effective accumulated temperature before and after silking (x2), rainfall before and after silking (x3), and sunshine hours before and after silking (x4) was y = 5465.19 + 17810.64x(1) - 23236.14x(2) + 4093.41x(3) + 6287.37x(4) (R2 = 0. 8187, P < 0.01), with the effects of these ecological factors on yield being in the sequence of x1 > x2 > x3 > x4 according to the absolute values of partial regression coefficients. In super high yielding (15499.86 kg x hm(-2)) spring maize population, the allocation ratios of x1, x2, x3, and x4 were 1.43, 1.41, 1.44, and 1.40, respectively. Therefore, in Northeast China, appropriate early sowing of spring maize to prolong its growth days with more rainfall and sunshine hours before silking could attain high yielding, and high or super high yield could be achieved when the allocation ratios of x1, x2, x3, and x4 were all about 1.4.

The objective of this study was to evaluate the efficacy and safety of the POF regimen (biweekly 5-fluorouracil/leucovorin combined with paclitaxel and oxaliplatin) as first-line treatment for advanced gastric cancer (AGC). Twenty-seven previously untreated patients with advanced adenocarcinoma of the gastric or gastroesophageal junction were eligible for this study. The chemotherapy regimen consisted of a 3-hour infusion of paclitaxel (135 mg/m(2)) followed by oxaliplatin (85 mg/m(2)) and leucovorin (400 mg/m(2)), administered simultaneously over a 2-hour infusion period, followed by an infusion of 5-fluorouracil (2400 mg/m(2)) over a 46-hour period. Twenty-one patients had measurable lesions: four complete responses, eight partial responses and seven stable diseases. At a median follow-up of 610 days, median survival was 348 days. Frequent grade 3 to 4 toxicities were: neutropenia (29.6%), stomatitis (7.4%), nausea (7.4%), vomiting (7.4%), hepatic dysfunction (3.7%), and fatigue (18.5%). No treatment-related deaths occurred. The POF regimen appears to be efficacious and is well tolerated in patients with AGC.

Objective: To establish an quick, sensitive and specific assay for effective inactivatian test of inactivated hepatitis A vaccine., Methods: effective inactivatian test of inactivated hepatitis A vaccine were carried out using integrated cell culture/strand-specific RT-PCR (ICC/strand-specific RT-PCR) assay compared with traditional ELISA and nest RT-PCR assay., Results: all the samples were infectious negative detecting by both ICC/ strand-specific RT-PCR and ELISA assay,while some samples appeared false positive detecting by nest RT-PCR., Conclusion: ICC/strand-specific RT-PCR assay is a novel, rapid, sensitive and reliable method for effective inactivatian test of inactivated hepatitis A vaccine. Shorting detection period largely, this assay may be used as an alternative method for routine inactivated hepatitis A vaccines test.

Objective: To establish ELISAs based on rLipL32/1-LipL21-OmpL1/2 fusion antigen of Leptospira interrogans for detecting specific IgG and IgM in serum of patients with leptospirosis., Methods: Microscope agglutination test(MAT) was performed to detect serum specimens from leptospirosis patients and to determine titers of rabbbit antiserum agaist rLipL32/1-LipL21-OmpL1/2 to reference standard strains of L. interrogans. By using rLipL32/1-LipL21-OmpL1/2, rLipL32/1, rLipL21 and rOmpL1/2 as the coated antigens, ELISAs for detecting specific serum IgM and IgG were established. The established ELISAs were applied to MAT-positive serum specimens from 107 patients with leptospirosis., Result: The results of MAT confirmed that 66% (71/107) of the patients were infected with L.interrogans serogroup Icterohaemorrhagiae, and the rLipL32/1-LipL21-OmpL1/2 antiserum were able to agglutinate all 15 reference standard L.interrogans strains with 1 : 20approximate, equals1 : 160 titers. The positive rates of ELISAs using rLipL32/1-LipL21-OmpL1/2, rLipL32/1, rLipL21 or rOmpL1/2 as the antigen were 89.7%, 75.7%, 85.1% and 79.4% for detecting IgM, respectively, while 99.1%, 99.1%, 94.4% and 86.0% for detecting IgG, respectively. The positive detection rate of rLipL32/1-LipL21-OmpL1/2-IgM-ELISA was higher than those of the other three IgM detection ELISAs (P<0.05). The positive detection rate of rLipL32/1-LipL21-OmpL1/2-IgG-ELISA was higher than that of rOmpL1/2-IgG-ELISA (P<0.05), while there was no significant differnce with that of rLipL21-IgG-ELISA and rLipL32/1-IgG-ELISA (P>0.05)., Conclusion: The ELISAs using rLipL32/1-LipL21-OmpL1/2 as the antigen can be applied as a sensitive,specific and universal serological method for diagnosis of leptospirosis.rLipL32/1-LipL21-OmpL1/2-IgM-ELISA shows a definite value for early diagnosis of leptospirosis compared with the other ELISAs used in this study.

Nitric oxide and nitric oxide synthases are key players in synaptic plasticity events in spinal cord (SC), which underlies the chronic pain states. To date, little is known about the molecular mechanisms regulating the activity of nitric oxide synthases in nociceptive systems. The present study was aimed at the determination of the gene expression of nNOS-interacting DHHC domain-containing protein with dendritic mRNA (NIDD), a recently identified protein regulating nNOS enzyme activity, in rat SC and dorsal root ganglia (DRG) and studying its regulation in states of nociceptive hypersensitivity in a rat model of neuropathic or inflammatory pain. It was found that NIDD mRNA was predominantly expressed in nociceptive primary neurons and in neurons of the spinal dorsal horn (DH) and the number of NIDD-positive neurons in the corresponding DRG or SC increased significantly following induction of chronic hyperalgesia. Meanwhile, remarkable changes of nNOS were detected under such pain conditions. Our data suggest a potential role for NIDD in the maintenance of thermal pain hypersensitivity possibly via regulating the nNOS activity.

Objective: To study the risk factors of tuberculosis in Yinchuan city and lay a basis for its prevention and control., Methods: A matched case-control (119:179) study for the risk factors was carried out. Data were analyzed with single-variable analysis and multiple factor logistic regression analysis., Results: Single-variable analysis showed that the education background (chi2 = 2.363, P = 0.018), family economic income (chi2 = 3.040, P = 0.002), smoking (chi2 = 2.500, P = 0.012), physical activities (chi2 = 2.330, P = 0.020), bacille Calmette-Guerin (BCG) vaccination history (chi2 = 22.151, P = 0.000), history of exposure to tuberculosis (chi2 = 15.740, P = 0.000) and so on had significant effects on tuberculosis. Multiple logistic regression analysis showed that family monthly income, smoking, physical activity, BCG vaccination history, history of exposure to tuberculosis entered the final regression model (chi2 = 5.880, 7.368, 3.891, 21.127, 14.536; OR = 0.529, 1.571, 0.774, 0.264, 3.978; P < 0.05)., Conclusion: History of exposure to tuberculosis and smoking should be the risk factors of tuberculosis in Yinchuan. Having much income, physical activities, and BCG vaccination history should be the preventive factors.

Objective: To design a new surgical correction and fixation technique for the treatment of severe angular kyphosis, observe the feasibility, safety and effectiveness of the technique., Methods: From May 2004 to February 2007, 16 cases with severe kyphosis (average 90.8 degrees, range 50 degrees-130 degrees) were treated with segmental resection osteotomy, section distraction, dual axial rotation correction and instrumentation fusion technique. The patients were inspected by local and total spine anteroposterior and lateral radiography pre-and postoperatively. The kyphotic Cobb angle was measured and 7 cases combined scoliosis Cobb angle was also measured. The Frankel Grading for neurological function, Oswestry Disability Index (ODI) and Patients Satisfactory Index (PSI) were evaluated preoperatively, postoperatively and at follow-up. The back pain relief was also observed., Results: The average surgical duration was 6.9 hours. The average blood loss was 4000 ml. The complications include 1 shifting of artificial vertebrae, 3 nerve root injury, 3 dural tear and 1 transitory dysfunction of lower extremity. All of these complications were relieved greatly after feasible treatment. The average follow-up time was 25 months. The average kyphotic angle was 90.8 degrees preoperatively, which was improved to 26.9 degrees immediately after surgery, and got an average correction rate of 72.5%. At follow-up, the average kyphotic angle was 28.9 degrees, and correction rate was 70.1%. The 7 cases who combined with scoliosis had an average Cobb angle of 35.9 degrees preoperatively, which decreased to 4.4 degrees immediately after surgery, and the correction rate was 87.2%. The correction rate was kept until follow-up (78.6%). Some patients got an improved neurological function. The Frankel Grading were E in 5 cases, D in 5 cases, C in 5 cases, and B in 1 case preoperatively. There were 10 cases of E grade, 3 cases of D grade, and 3 cases of C grade at follow-up. Except 3 cases who had no symptoms before surgery, the average ODI was 18.9 preoperatively, and 10.8 postoperatively. The average improvement of ODI was 52.7%. The PSI result showed a satisfied rate of 93.8%. The back pain of 3 cases were totally relieved after surgery., Conclusions: Segmental resection osteotomy with dual axial rotation correction and fusion technique is an effective way to treat severe angular kyphosis. It is a safe technique and has high correction rate. The long-term results is acceptable.

Objective: To determine the location on outer envelope and natural antibody response and types of genus-specific lipoprotein antigen LipL41s in patients with Leptospira interrogans., Methods: Microscope agglutination test (MAT) was used to examine leptospirosis patients' serum samples from Sichuan area, China. Ni-NTA affinity chromatography was performed to extract the target recombinant rLipL41/1 and rLipL41/2 products that expressed under inducement of IPTG. Western blot assay was performed to detect the immunoreactivity between the sera from the patients infected with different serogroups of L. interrogans and rLipL41s. Immune aurosol electron microscopy was selected to locate the position of LipL41s on leptospiral envelope. ELISA based on rLipL41s was established to confirm the level and types of specific antibody., Results: L. interrogans serogroup icterohaemorrhagiae remained to be the most dominant leptospiral serogroup in Sichuan area. All the sera from patients infected with different serogroups of L. interrogans could efficiently recognize the LipL41s which were the protein molecular that located on the external surface of leptospiral envelope. In the 156 serum samples from MAT positive leptospirosis patients, the positive rates for rLipL41/1 or rLipL41/2 specific IgM appeared to be 84.6%-87.8% and 78.2%-83.3%, respectively, while for rLipL41/1 or rLipL41/2 specific IgG they were 69.2%-81.4% and 75.0%-80.1%, respectively., Conclusion: LipL41s were the leptospiral superficial protein antigen of L. interrogans. Both the LipL41/1 and LipL41/2 could induce serum antibodies IgM and IgG with extensive antigenic-cross reaction during natural infection of L. interrogans in general populations. Hence, rLipL41/1 or rLipL41/2 could be used as the antigen candidate for developing universal genetic engineering vaccine and detection kit.

Objective: To establish a standardized operation procedure for pulsed-field gel electrophoresis (PFGE) on Leptospira interrogans as well as a figure digital database to develop the Chinese representative reference strains., Methods: Under the characteristics of strains and referring to the other SOPs of PFGE on pathogens provided by CDC and PulseNet Asia Pacific, genomic chromosome DNA purification, restriction endonuclease digestion and the parameters for running PFGE were optimized., Results: Not I digestion patterns of leptospiral genome for the Chinese representative strains were established and partial isolates of serogroup icterohaemorrhagiae from the leptospirosis surveillance in Sichuan and Anhui provinces were analyzed by PFGE. Results showed that each of all the 15 Chinese representative strains had a unique pattern. 91.67% (22/24) of the 24 isolates identified as serogroup icterohaemorrhagiae matched to the map of the reference strain 56601 (serogroup icterohaemorrhagiae serovar lai)., Conclusion: The PFGE figures were clear with high resolution and the fragments were equally distributed by this standardized operating procedure so as to reveal the molecular-genetic characteristics of Leptospira interrogans. The patterns had high relativity with the serological identification and seemed to be very important for genetic analysis of strains in studying the outbreak of leptospirosis.

Objective: To produce specific monoclonal antibody (mAb) against recombinant human erythropoietin (rHuEPO) for development of highly efficient methods for erythropoietin detection in biological fluids., Methods: rHuEPO was covalently coupled with bovine serum albumin (BSA) and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology. The obtained F3-mAb was characterized by enzyme-linked immunosorbent assay (ELISA), SDS-PAGE and Western blot., Results: The isotype of F3-mAb was found to be IgM with an affinity constant of 2.1 x 10(8) L/mol. The competitive ELISA using the obtained IgM showed a broader linear range and lower detection limit compared with previous work., Conclusions: The modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.

Aim: To investigate whether myosin activated-T lymphocytes induce myocardial autoimmune response., Methods: Dendritic cells and T lymphocytes were isolated and purified from spleens of Lewis rats. After cocultured in the absence or presence of cardiac myosin (20 mug/mL), the sensitized or unsensitized T lymphocytes were transferred into the naive syngeneic Lewis rats by a single tail vein infusion, respectively. The recipient rats were killed on the third day, at the first and fourth week and then the histopathological changes of the rats were observed., Results: T lymphocyte infiltration was observed on the third day. It reached the peak with occasional myocardium necrosis at the first weekend. T cell infiltration distinctly lessened at the fourth weekend. No T lymphocyte infiltration was found in kidney, liver, lung, and brain. T lymphocyte infiltration wad not detected in the heart of the transferred unsensitized T lymphocytes of the rats., Conclusion: Myocardial autoimmune response can be induced by myosin activated T lymphocytes.

The major aim of this study is to determine the location on outer envelope of the genus-specific antigen OmpL1s of Leptospira interrogans, and the inducement of naturally antibody response and types of the antigen, which will offer the evidences to use OmpL1s as the antigen candidate for developing universal genetic engineering vaccine and detection kit. The serum samples from 156 leptospirosis patients in Sichuan area were detected using microscope agglutination test (MAT). By using PCR plus nucleotide sequence analysis, the genotypes of the dominant L. interrogans serogroups in China were demonstrated. Routine genetic engineering technique was applied to construct the prokaryotic expression systems of genotypes ompL1/1 and ompL1/2, and Ni-NTA affinity chromatography was performed to extract the target recombinant products rOmpL1/1 and rOmpL1/2. Immune aurosol electron microscopy was selected to locate the position of OmpL1s on leptospiral envelope. ELISAs based on rOmpL1s were established to confirm the level and types of specific antibody. The results indicated that L. interrogans serogroup Icterohaemorrhagiae remains to be the most important dominant leptospiral serogroup in Sichuan area. There are two ompL1 genotypes of ompL1/1 and ompL1/2 in the dominant leptospiral serogroup in China. And remarkable differences of the nucleotide and putative amino acid sequence similarities between the two genotypes are present. OmpL1s are the protein molecular that located on the external surface of leptospiral envelope. In the 156 cases of leptospirosis patients' serum samples using different dilutions, the positive rates for rOmpL1/1 or rOmpL1/2 specific IgM are 67.9%-79.5% and 75.0%-75.6%, while for for rOmpL1/1 or rOmpL1/2 specific IgG are 71.8%-79.5% and 75.0%-76.9%, respectively. All the results mentioned above lead to the conclusions that OmpL1s is the leptospiral genus-specific superficial protein antigen of L. interrogans and both rOmpL1/1 and rOmpL1/2 can induce humoral response in individuals naturally infected with L. interrogans as well as produce two serum antibodies IgM and IgG with extensive antigenic-cross reaction. Therefore, rOmpL1/1 and rOmpL1/2 can be used as the antigen candidate for developing universal genetic engineering vaccine and detection kit.

Metallothionein (MT), a metal-binding protein induced primarily by heavy metals in vertebrates, is considered a biomarker for environmental heavy-metal contamination. To investigate heavy metal pollution in the freshwater environment, MT-I and MT-II were purified from livers of crucian carp (Carassius carassius) by gel exclusion chromatography and ion exchange chromatography. To detect the purified MT-II, a specific monoclonal antibody (mAb) against crucian carp MT-II was produced from the hybridoma strains by cell-cell fusion. By using Enzyme-Linked Immunosorbent Assay (ELISA) with this mAb, the purified crucian carp MT-II was detected with a high specificity and sensitivity. There was a good correlation between the amount of MT-II in carp livers and the concentration of heavy metals in water. ELISA was then used to evaluated the degree of heavy metal pollution in two freshwater systems. The results indicate that the MT-II content in carp liver tissue can be used as an indicator of environmental heavy-metal pollution.