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Background: The COVID-19 pandemic is a significant health threat. Health care worker (HCWs) are at a significant risk of infection which may cause high levels of psychological distress. The aim of this study was to investigate the psychological impact of the COVID-19 on HCWs and factors which were associated with these stresses during the first outbreak in Shanghai.Methods: Between February 9 and 21, 2020, a total of 3,114 frontline HCWs from 26 hospitals in Shanghai completed an online survey. The questionnaire included questions on their sociodemographic characteristics, 15 stress-related questions, and General Health Questionnaire-12 (GHQ-12). Exploratory factor analysis was applied to the 15 stress-related questions which produced four distinct factors for evaluation. Multiple linear regression models were performed to explore the association of personal characteristics with each score of the four factors. Binary logistic analysis was used to explain the association of personal characteristics and these four factors with the GHQ-12.Results: There were 2,691 valid surveys received. The prevalence of emotional distress (defined as GHQ-12 ≥ 12) was noted in 47.7% (95%CI:45.7–49.6%) HCWs. Females (OR = 1.43, 95%CI:1.09–1.86) were more likely to have a psychological distress than males. However, HCWs who work in secondary hospitals (OR = 0.71, 95% CI:0.58–0.87) or had a no contact history (OR = 0.45, 95%CI: 0.35–0.58) were less likely to suffer psychological distress. HCWs who were nurses, married, and had a known contact history were highly likely to have anxiety. HCWs working at tertiary hospitals felt an elevated anxiety regarding the infection, a lack of knowledge, and less protected compared to those who worked at secondary hospitals.Conclusions: Our study shows that the frontline HCWs had a significant psychosocial distress during the COVID-19 outbreak in Shanghai. HCWs felt a lack of knowledge and had feelings of being not protected. It is necessary for hospitals and governments to provide additional trainings and psychological counseling to support the first-line HCWs.

Summary Background The outbreak of COVID-19 has led to international concern. We aimed to establish an effective screening strategy in Shanghai, China, to aid early identification of patients with COVID-19. Methods We did a multicentre, observational cohort study in fever clinics of 25 hospitals in 16 districts of Shanghai. All patients visiting the clinics within the study period were included. A strategy for COVID-19 screening was presented and then suspected cases were monitored and analysed until they were confirmed as cases or excluded. Logistic regression was used to determine the risk factors of COVID-19. Findings We enrolled patients visiting fever clinics from Jan 17 to Feb 16, 2020. Among 53 617 patients visiting fever clinics, 1004 (1·9%) were considered as suspected cases, with 188 (0·4% of all patients, 18·7% of suspected cases) eventually diagnosed as confirmed cases. 154 patients with missing data were excluded from the analysis. Exposure history (odds ratio [OR] 4·16, 95% CI 2·74–6·33; p

IntroductionAcute exacerbation (AE) is a major cause of disease progression and death in patients with chronic obstructive pulmonary disease (COPD), accounting for majority of medical expenditures. Correct inhalation therapy is effective in preventing AE attacks. However, inappropriate usage of dry powder inhaler, partially due to the unrecovered peak inhalation flow rate (PIFR) after acute exacerbation of COPD (AECOPD), results in increased risk of early treatment failure. Therefore, we designed a multicentre, randomised clinical trial to determine whether PIFR-based optimised inhalation therapy and training on inhaler usage at discharge could effectively reduce early treatment failure events.Methods and analysisA total of 416 hospitalised patients just recovering from AECOPD will be recruited and equally randomised into the PIFR group and the control group at a 1:1 ratio. The PIFR group will receive additive support before discharge, including choice of PIFR-guided inhaler and education on its usage. PIFR is measured by InCheck DIAL. In comparison, the control group will receive inhalers based on judgement of the respiratory physician. The primary outcome of the study is 30-day treatment failure rate. Other endpoints include PIFR, error rate of inhalation device use, satisfaction with inhalation devices, 30-day mortality, 90-day mortality, symptoms and quality of life of patients, and COPD-related treatment costs.Ethics and disseminationThe trial has been approved by the Ethics Committee of Zhongshan Hospital of Fudan University (B2019-142). Participants will be screened and enrolled from hospitalised patients with AECOPD by clinicians, with no public advertisement for recruitment. After the trial has completed, the results will be reported to the public through conference presentations and peer-reviewed journals.Trial registration numberNCT04000958.

Background: The outbreak of novel coronavirus disease 2019 (COVID-19) has led to increasing international concern. Effective screening strategy and early recognition of the disease are crucial to limiting spread of this worldwide challenge. Methods: Fever clinics of 25 hospitals evenly distributed in 16 Districts of Shanghai were selected. All patients visiting the fever clinics from the begin of the outbreak of Shanghai (17 January 2020) to 16 February were enrolled. A standardized procedure for COVID-19 screening was presented and all the suspected cases were monitored and analyzed till they were confirmed or excluded. Findings: Among 53617 patients who visiting fever clinics, 1004 (1·87%) were considered as suspected cases, with 188 (0·35% of all patients, 18·73% of suspected cases) diagnosed as confirmed cases finally. Compared with the excluded cases, confirmed cases were older and the highest temperatures were relatively lower (p

Bluetooth technology is traditionally based on the point-to-point transmission protocol. It has limited transmission distance and poor networking capabilities, making it is very rare in the field of Internet of things. Traditional switches are mechanical switches, which are not safe or convenient. The traditional way to achieve this is to use Bluetooth devices and microcontrollers implement one-on-one control. Based on these issues, we will propose the concept of Bluetooth Mesh. It is a multi-hop technology that extends the transmission distance between Bluetooth nodes in a wide range and solves the limitations of traditional Bluetooth. By embedding Bluetooth Mesh technology into each switch, a Bluetooth Mesh network system is formed, and the system can be controlled by the mobile phone. The Bluetooth Mesh network switch control system can integrate technology into life and make people's life more secure and convenient.

MapReduce has been shown promising to harness the multicore platform. Existing MapReduce libraries on multicore are written with shared-memory Pthreads, which introduce pervasive nondeterminism and might produce nondeterministic results if user-provided map or reduce functions are sensitive to the input order. We propose DMR, a deterministic MapReduce library, to ensure deterministic program behaviors no matter whether map/reduce function is sensitive to the input order. DMR adopts a round-robin scheduling of map tasks and a partitioned scheduling of reduce tasks to ensure deterministic scheduling. DMR is written with a deterministic message passing multithreaded model (DetMP) to provide Phoenix-like API, thus Phoenix workloads can be built and run on DMR with no or little change. Evaluation results by testing seven Phoenix workloads show that DMR only runs worse than Phoenix on an iterative MapReduce application kmeans, outperforms Phoenix between 1.42X and 3.33X faster on pca and word_count, and scales better than Phoenix on 3 of the rest 4 workloads.

We retrospectively reviewed the medical records of 17 fatal H7N9 cases in Shanghai in 2013, analyzed clinical variables and described their clinical and epidemiologic characteristics. The median age was 73 years, and 82.4% had underlying medical conditions. The most frequent symptoms were fever (100%), followed by productive cough (47.1%) and dry cough (35.5%). Thirteen (76.5%) patients had dyspnea or respiratory distress, five (29.4%) had shock, and four (23.5%) had acute kidney injury. Seventeen (100.0%) patients had lymphopenia. Involvement of both lungs was found by chest radiography in 14 (82.4%) patients at presentation. Fifteen (88.2%) patients were hospitalized. The median times from illness onset to hospitalization and to diagnosis confirmation were both six days. Eleven (64.7%) patients were admitted to the intensive care unit. Sixteen (94.1%) patients were treated with oseltamivir. The median time from illness onset to oseltamivir treatment was six days. Among six patients for whom the duration of viral shedding was available, the median duration of viral shedding after oseltamivir treatment was 17 days. The median time from illness onset to death was 11 days. Refractory hypoxemia accounted for most deaths. The clinical and epidemiologic characteristics in the Shanghai fatal series of patients do not differ from other reports of H7N9 patients in China. This investigation reflects a delay in the diagnosis and antiviral treatment of H7N9 patients in the early stage of the epidemic in Shanghai. Late antiviral treatment and a long duration of viral shedding may be associated with a fatal outcome in these patients.

Although deletions or inactivating mutations of the tumor suppressor gene PTEN (phosphatase and tensin homolog deleted on chromosome 10) are involved in the development of a variety of tumors including glioblastoma, melanoma, prostate cancer, breast cancer, endometrial cancers etc., the role of PTEN expression in human primary hepatocellular carcinoma (HCC) has not yet been clarified. The aim of this study is to investigate the involvement of PTEN mRNA and protein expression in HCC. The level of PTEN mRNA expression in HCC specimens was analyzed by Northern blot. PTEN poly-clonal antibody was raised by immunizing New Zealand white rabbit with (His)6-tagged PTEN fusion protein and characterized by Western blot. The level of PTEN protein expression was determined by immunohistochemistry. The significance of PTEN in HCC was analyzed by comparing its expression level with the clinicopathological parameters of HCC patients. Four transcripts of PTEN mRNA at 5.5 kb, 4.4 kb, 2.4 kb, and 1.8 kb were detected in most para-carcinoma liver tissues, and the expression level of PTEN mRNA in carcinoma liver tissues was found to decrease significantly. The poly-clonal antibody raised against histidine-tagged fusion PTEN protein showed specific immuno-reactivity to PTEN protein. Using the specific poly-clonal antibody prepared and characterized by ourselves, we found that PTEN protein was significantly down-regulated in HCC tissues compared with paired para-carcinoma tissues. The protein expression of PTEN is negatively associated with the pathological grading and presence of cancer thrombus of HCC. Down-regulation of PTEN expression may play an important role in the development of HCC and the level of PTEN expression may be a potential adjuvant parameter in forecasting the progression and prognosis of HCC patients.

To observe the impacts of acupuncture on intelligent structure, social adaptability and fMRI brain function in children mental retardation (MR).Sixty cases of MR in compliance with the diagnostic standard were randomized into an acupuncture group and a medication group, 30 cases in each one. In the acupuncture group, Sishenzhen [four points, 1.5 cun anterior, posterior and bilateral to Baihui (GV 20)], Zhisanzhen [Shenting (GV 24), bilateral Benshen (GB 13)], Niesanzhen (the point 2 cun directly above the ear a-pex, the two points 1 cun bilateral the first point) and Naosanzhan [Naohu (GV 17) and bilateral Naohu (GB 19)] were selected as the main points. In the medication group, piracetam tablets were prescribed for oral administration. One course of treatment was 4 months in the two groups. The comprehensive efficacy was compared between the two groups at the end of treatment course. China-Wechsler Intelligence Scale for Children (C-WISC) was used to assess the intelligent improvements. Infant-Junior School Student Social Life Ability Scale was adopted to assess the improvements of social adaptability. Five cases were selected from the acupuncture group and fMRI was adopted to compare the brain function imaging changes before and after acupuncture treatment.In the acupuncture group, the final intelligence quotient (FIQ) and social adaptability score after treatment were higher than those before treatment (P0.05), of which, the performance intelligence quotient (PIQ) was improved significantly, indicating the statistically significant difference (P0.05). But the verbal intelligence quotient (VIQ) did not change apparently (P0.05). In the medication group, the changes in all the indices were not apparent before and after treatment (P0.05). In comparison of the changes after treatment between the two groups, FIQ, PIQ and social adaptability score in the acupuncture group were improved more significantly as compared with the medication group (P0.05). The fMRI brain function images did not change apparently before and after treatment in those 5 cases of the acupuncture group.Acupuncture promotes the intelligent recovery of MR children and improves their social adaptability. It indicates the satisfactory clinical efficacy. But, the fMRI brain function images do not change apparently before and after treatment.

During the spring of 2013, a novel avian-origin influenza A (H7N9) virus emerged and spread among humans in China. Data were lacking on the clinical characteristics of the infections caused by this virus.Using medical charts, we collected data on 111 patients with laboratory-confirmed avian-origin influenza A (H7N9) infection through May 10, 2013.Of the 111 patients we studied, 76.6% were admitted to an intensive care unit (ICU), and 27.0% died. The median age was 61 years, and 42.3% were 65 years of age or older; 31.5% were female. A total of 61.3% of the patients had at least one underlying medical condition. Fever and cough were the most common presenting symptoms. On admission, 108 patients (97.3%) had findings consistent with pneumonia. Bilateral ground-glass opacities and consolidation were the typical radiologic findings. Lymphocytopenia was observed in 88.3% of patients, and thrombocytopenia in 73.0%. Treatment with antiviral drugs was initiated in 108 patients (97.3%) at a median of 7 days after the onset of illness. The median times from the onset of illness and from the initiation of antiviral therapy to a negative viral test result on real-time reverse-transcriptase-polymerase-chain-reaction assay were 11 days (interquartile range, 9 to 16) and 6 days (interquartile range, 4 to 7), respectively. Multivariate analysis revealed that the presence of a coexisting medical condition was the only independent risk factor for the acute respiratory distress syndrome (ARDS) (odds ratio, 3.42; 95% confidence interval, 1.21 to 9.70; P=0.02).During the evaluation period, the novel H7N9 virus caused severe illness, including pneumonia and ARDS, with high rates of ICU admission and death. (Funded by the National Natural Science Foundation of China and others.).

The pancreatic and duodenal homeobox factor-1 (Pdx1) is essential for pancreatic development and insulin gene transcription, whereas c-Myc has a deleterious effect on islet function. However, the relationship between c-Myc and Pdx1 is poorly concerned. Here we demonstrated that Pdx1 could suppress c-Myc promoter activity, which relied on T cell factor (Tcf) binding elements harbored in c-Myc promoter. Furthermore, the transcription activity of β-catenin/Tcf was markedly decreased on Pdx1 expression, but cotransfection of Pdx1 short hairpin RNA abrogated this effect. Pdx1 expression did not induce β-catenin degradation nor did it alter their subcellular distribution. The mutation analysis showed that the amino acids (1–209) of Pdx1 harboring an inhibitory domain, which might lead to the reduction of β-catenin/Tcf/p300 complex levels and attenuate their binding activity with c-Myc promoter sequences. Moreover, adenovirus-mediated Pdx1 interference caused cell proliferation and cytokine-induced apoptosis via the dysregulation of c-Myc transcription. These results indicated that the Pdx1 functioned as a key regulator for maintenance of β-cell function, at least in part, through controlling c-Myc expression and the loss of its regulatory function may be an alternative mechanism for β-cell neogenesis and apoptosis found in diabetes.

Signal regulatory protein (Sirp) is a recently isolated, cloned and identified inhibitor receptor distributed in the membrane of hematopoietic and nonhematopoietic cells. Sirp alpha1 (Sirpalpha1) is a member of Sirp families. Sirpalpha1 can bind SHP-2 in the form of tyrosine phosphorylation by SH2 effect and negatively regulate growth factor, oncogene, or insulin-induced responses as its substrate. This study aimed to preliminarily clarify the negatively regulating proliferation mechanism of Sirpalpha1 in liver cancer.pLXSN, Sirpalpha1 and Sirpalpha1P4Y2 plasmids were respectively transfected into Sk-Hep1 liver cancer cell line, and various stable Sk-Hep1 cell lines were obtained with screening agent of G418 (1200 microg/ml). The expressing levels of cyclin D1, CDK4, Fas, beta-catenin and gankyrin in various cell lines were determined with Western blotting. Cell cycles were determined at 0, 12 and 24 hours with flow cytometry after various synchronous cell lines were cultured without serum for 72. Cell apoptosis induced with agent of TNF-alpha (50 ng/ml) was determined with flow cytometry at 0, 0.5, 1, 3, 6 and 12 hours.Sirpalpha1 could significantly decrease the expression of cyclin D1, beta-catenin and gankyrin, but it couldn't affect the expression level of CDK4 and Fas. When synchronous cells were cultured for 12 hours, S phase Sk-Hep1 cell transfected with Sirpalpha1 plasmid was the lowest [(31.92+/-0.22)% vs. other cell lines, P0.05], and the cell line was highly sensitive to TNF-alpha agent for 1 hour. (59.31+/-0.59)% of apoptotic cells occurred (vs. the other time points, P0.05).Sirpalpha1 might block the cell cycle of liver cancer, inhibit cell proliferation, promote cell apoptosis by decreasing the expression of cyclin D1, beta-catenin and gankyrin. It is one of the important mechanisms inhibiting the occurrence and development of hepatocellular carcinoma.

beta-Catenin is a key molecule involved in both cell adhesion and Wnt signaling pathway. However, the exact relationship between these two roles has not been clearly elucidated. Tyrosine phosphorylation of beta-catenin was shown to decrease its binding to E-cadherin, leading to decreased cell adhesion and increased beta-catenin signaling. We have previously shown that receptor-like protein-tyrosine phosphatase PCP-2 localizes to the adherens junctions and directly binds and dephosphorylates beta-catenin, suggesting that PCP-2 might regulate the balance between signaling and adhesive beta-catenin. Here we demonstrate that PCP-2 can inhibit both the wild-type and constitutively active forms of beta-catenin in activating target genes such as c-myc. The phosphatase activity of PCP-2 is required for this effect since loss of catalytic activity attenuates its inhibitory effect on beta-catenin activation. Expression of PCP-2 in SW480 colon cancer cells can lead to stabilization of cytosolic pools of beta-catenin perhaps, by virtue of their physical interaction. PCP-2 expression also leads to increased membrane-bound E-cadherin and greater stabilization of adherens junctions by dephosphorylation of beta-catenin, which could further sequester cytosolic beta-catenin and thus inhibit beta-catenin mediated nuclear signaling. Furthermore, SW480 cells stably expressing PCP-2 have a reduced ability to proliferate and migrate. Thus, PCP-2 may play an important role in the maintenance of epithelial integrity, and a loss of its regulatory function may be an alternative mechanism for activating beta-catenin signaling.

Background: Human gankyrin gene product (p28GANK) is a novel oncogenic protein ubiquitously overexpressed in hepatocellular carcinoma and also plays a role in cell cycle progression in normal hepatocytes and liver regeneration. However, little is known about the physiological role of p28GANK in the liver oval cell activation and proliferation. We investigated the possible involvement of p28GANK in oval cell-mediated liver regeneration and cell cycle progression. Methods: We examined the different p28GANK expression in 2-acetylaminofuorene/partial heptectomy (2-AAF/PH) rats, as a model of oval cell activation, and PH rats by Western blot and immunohistochemistry. Oval cells isolated from 2-AAF/PH rat model were cultured in our study. p28GANK expression was examined in the oval cells after mitogenic stimulation. Results: In 2-AAF/PH rats, p28GANK was expressed in the activated oval cells and located in the nucleus. p28GANK protein expression was increased in 2-AFF/PH rats after hepatectomy lasting for 96 h when retinoblastoma maintained hyperphosphorylation status at Ser-795. The isolated oval cells express AFP, OV6, CK19, CD34, CD45, c-kit and albumin. After epidermal growth factor stimulation, p28GANK protein was up-regulated in oval cells from 24 to 72 h, which coincided with increased expression of CyclinD1, CDK4 and decreased of Rb protein. Conclusions: p28GANK expression was increased in oval cell-mediated liver regeneration and oval cells after mitogenic stimulation. Thus, p28GANK may play a role in oval cell-mediated liver regeneration and liver oval cell cycle progression.

Background & Aims: RNA interference (RNAi) is a powerful tool to silence gene expression. The adenoviral vector expressing small interfering RNA (siRNA) is highly effective in mammalian cells. However, its potential use as a therapeutic tool to target an oncogene specifically remains to be seen. We applied the adenovirus-delivered siRNA (AdSiRNA) to inhibit a hepatocellular carcinoma (HCC) oncogene, p28 GANK , in HCC cell lines and investigated its antitumor effects. Methods: The T7-RNA polymerase system was used to screen the specific target site. Double-strand oligonucleotide for transcription of short hairpin RNA was constructed into the adenoviral vector. Four HCC cell lines were infected with the RNAi-containing adenovirus. The RNAi effects on HCC were studied in cultured cells as well as in animal models. Results: p28 GANK expression was suppressed by up to 80% in HCC cells. Depletion of p28 GANK inhibited HCC cell growth and tumorigenesis, enhanced dephosphorylation of RB1, and decreased transcription activity of E2F-1 in HuH-7 cells. Furthermore, depletion of p28 GANK induced caspase-8- and caspase-9-mediated apoptosis of HCC cells. Finally, targeting p28 GANK by adenovirus injection inhibited the growth of established tumors in nude mice. Conclusions: This study shows that the T7-system screening-based AdSiRNA can be used successfully to silence an oncogene. We proved the therapeutic potential of AdSiRNA on the treatment of HCC by targeting p28 GANK . Our results indicate that p28 GANK may serve as a novel therapeutic target for treating HCC.

Signal regulatory protein alpha1 (Sirpalpha1) is a negative regulatory factor, and inhibits receptor tyrosine kinase-dependent cell proliferating signal. This study was undertaken to observe the effect of signal regulatory proteinalpha1 (Sirpalpha1)on gankyrin,cyclin D1,CDK4 and Fas expression in Sk-hep1 mouse hepatoma carcinoma cell line.BOSC 23 packed cells were respectively transfected by means of recombinated retrovirus including pLXSN, pLXSN- Sirpalpha1 and pLXSN- Sirpalpha1P4Y2 with lipofectin, and various plasmid virus media(viral titer 2.1X10(6) CFU/ml) were collected and infected respectively in 80% confluent Sk-hep1 cells. Transfected Sk-hep1 cells were selectively screened with G418 (1200 mug/ml), and Sk-hep1 cell lines transfected with various plasmids were obtained. The protein expressions of gankyrin, cyclin D1, CDK4 and Fas in various Sk-hep1 lines were determined by Western blotting. Various Sk-hep1 lines were recovered to culture with 10% fetal bovine serum at 12 hours and 24 hours after starving culture with free serum for 72 hours, and cells were collected to determine the percentage of S phase cells of proliferating cycle by flow cytometry.Sirpalpha1 transfection remarkably downregulated gankyrin and cyclin D1 expression. Sirpalpha1P4Y2 downregulation of gankyrin expression was greater than that of Sirpalpha1 (P0.05), but no significant effect of Sirpalpha1 and Sirpalpha1P4Y2 on CDK4 and Fas protein expression was observed in transfected Sk-hep1 lines (P0.05). The percentage of S phase cells significantly decreased in Sk-hep1 cells transfected with Sirpalpha1 and Sirpalpha1P4Y2 plasmids (vs pLXSN Sk-hep1, P0.05). The percentage of S phase cells in various Sk-hep1 cells increased when recovering to culture with 10% fetal bovine serum at 12 hours, but the percentage of S phase cells in Sk-hep1 cells transfected with Sirpalpha1 was the lowest (vs pLXSN and Sirpalpha1P4Y2 Sk-hep1, P0.05). The percentage of S phase cells in transfected pLSXN Sk-hep1 cells was the largest (vs Sirpalpha1 and Sirpalpha1P4Y2 Sk-hep1, P0.05). There was no significant difference between the transfected Sirpalpha1 Sk-hep1 cells and Sirpalpha1P4Y2 Sk-hep1 cells (P0.05).Sirpalpha1 decreases gankyrin and cyclin D1 expression, and inhibits proliferation of liver carcinoma cells. It may be one of the forms for an Sirpalpha1 negative regulation of carcinogenesis and development of hepatocellular carcinoma.

Signal regulatory protein (SIRP) alpha1 is a member of the SIRP family that undergoes tyrosine phosphorylation and binds SHP-2 tyrosine phosphatase in response to various mitogens. The expression levels of SIRPalpha1 were decreased in HCC tissues, compared with the matched normal tissues. Exogenous expression of wild type SIRPalpha1, but not of a mutant SIRPalpha1 lacking the tyrosine phosphorylation sites, in SIRPalpha1-negative Huh7 human HCC cells resulted in suppression of tumor cell growth both in vitro and in vivo. Treatment of Huh7 transfectants with EGF or HGF induced tyrosine phosphorylation of SIRPalpha1 and its association with SHP-2, which were accompanied by reduced ERK1 activation. Expression of SIRPalpha1 significantly suppressed activation of NF-kappaB and also sensitized Huh7 cells to TNFalpha or cisplatin-induced cell death. In addition, SIRPalpha1-transfected Huh7 cells displayed reduced cell migration and cell spreading in a fashion that was dependent on SIRPalpha1/SHP-2 complex formation. In conclusion, a negative regulatory effect of SIRPalpha1 on hepatocarcinogenesis is exerted, at least in part, through inhibition of ERK and NF-kappaB pathways.

Signal regulatory protein alpha (SIRPalpha) is an Ig superfamily protein whose cytoplasmic region contains immunoreceptor tyrosine-based inhibitory motif (ITIM), which when tyrosine phosphorylated binds the SH2-domain containing phosphatase 2 (SHP-2). Both SIRPalpha and SHP-2 are highly expressed in brain. Murine cerebellar cells cultured on SIRPalpha-coated surface exhibit enhanced neurite outgrowth and SIRPalpha is localized at sites of synaptogenesis in postnatal mouse brain. In this study, we show that nerve growth factor (NGF) stimulation resulted in elevated SIRPalpha expression during PC12 differentiation. We also show that NGF-induced morphological differentiation, but not growth arrest response, was inhibited by ectopic SIRPalpha expression. PC12 cells stably expressing SIRPalpha proliferated more rapidly than mock-transfected cells. The activity of c-jun N-terminal kinase (JNK) decreased in SIRPalpha-transfected PC12 cells, whereas nuclear factor-kappaB (NF-kappaB) activity increased. Collectively, our results suggest that SIRPalpha may stabilize synaptic connections by inhibiting improper neurite outgrowth and might realize its neuronal function, at least in part, by modulating JNK and NF-kappaB activity.

To explore the significance of PTEN (phosphatase and tensin homolog deleted on chromosome 10) in the development of human primary hepatocellular carcinoma (HCC).PTEN protein expression in cancerous liver tissues and paired para-carcinoma liver tissues from 60 HCC patients was detected by immunohistochemistry and PTEN mRNA expression was analyzed by northern blot. The significance of PTEN in the development of HCC was analyzed by investigating the relationship between the expression levels of PTEN protein and mRNA, and the clinicopathological parameters of HCC patients.PTEN protein was immunohistochemically stained in the cytoplasmic region of para-carcinoma liver tissues in all the 60 patients, while only 48.3% (29/60) of the patients were positive for PTEN protein in cancerous liver tissues. The positive rate of PTEN protein in HCC tissues were relative to the histological gading and the presence of tumor thrombus. In grade I - II, III, and IV, the positive rates were 84.0%, 23.8%, and 21.4% respectively, and in the group with tumor thrombus was 26.7%, while in the group without tumor thrombus was 55.6%. Northern blot showed that there existed four PTEN mRNA transcripts with the length of 5.5kb, 4.4kb, 2.4kb, and 1.8kb respectively. The level of PTEN mRNA expression in HCC tissues was much lower than that in the paired para-carcinoma liver tissues. The low expression level of the 5.5kb and 4.4kb transcripts was significantly associated with serum AFP value, presence of tumor thrombus, state of satellite lesion and histological grading. The low expression level of the 2.4kb transcript in HCC was significantly associated with the presence of tumor thrombus and satellite lesions in HCC patients. However, no evident relationship between the lowered expression level of the 1.8kb transcript and the clinicopathological parameters of HCC was observed in these 60 patients.Down-regulation of PTEN expression may play an important role in the development of HCC and the expression level of PTEN may be a potential adjuvant parameter in forecasting the progress and prognosis of HCC.

To study the inhibitory effect of retroviral packaging cells injected intrasplenically encoding mouse interleukin-12 (mIL-12) and human interleukin-2 (hIL-2) fusion gene on the growth of hepatocellular carcinoma.The retroviral vectors encoding mIL-12 gene, hIL-2 gene, and mIL-12 and hIL-2 genes, GCIL12EXPN, GCXEIL2PN, and GCIL12EIL2PN were constructed and then transfected into the retroviral packaging cells PA317 to construct cells PA317-GCIL12EXPN, PA317-GCXEIL2PN, and PA317-GCIL12EIL2PN. Rat hepatocellular carcinoma cells CBRH3 were implanted into the livers of Wistar rats to establish hepatoma animal model. Then the rats were divided into 5 groups to be injected intrasplenically with normal saline one day after the implantation (0.8 ml/rat, group I, n = 10), blank vector PA317-GCXEXPN one day after the implantation (10(7) cells/rat, group II, n = 10), PA317-GCIL12EXPN containing IL-2 gene (1 x 10(7) cells/rat 1, 3, 5, or 7 days after the implantation, group III, n = 40), PA317-GCXEIL2PN containing mIL-12 gene (1 x 10(7) cells/rat 1, 3, 5, or 7 days after the implantation, group IV, n = 40), and PA317-GCIL12EIL2PN containing IL-12-IL-2 fusion gene (1 x 10(7) cells/rat 1, 3, 5, or 7 days after the implantation, group V, n = 40) respectively. The rats surviving longer than 2 months were re-injected with hepatocellular carcinoma cells. The therapeutic effect, immune function and toxic effect were evaluated. CT was conducted on the liver before and after the experiment. Laparotomy was performed 3 and 7 days after treatment to resect some of the carcinoma tissues to undergo pathological examination and OX8 immunohistostaining. Serum mIL-12 and hIL-2 were detected one day before and 3, 7, 30, and 60 days after treatment.The average survival times of the rats treated with IL-12-IL-2 fusion gene at the first, third, fifth and seventh day after tumor implantation were 53.3 +/- 3.7 days, 49.3 +/- 4.2 days, 31.0 +/- 2.1 days, and 24.3 +/- 1.4 days respectively, longer than those treated with IL-2 gene (25.0 +/- 2.5 days, 23.5 +/- 2.0 days, 18.3 +/- 2.4 days, and 12.0 +/- 1.8 days respectively, P0.001), and those treated with IL-12 gene (39.0 +/- 4.8 days, 32.0 +/- 3.9 days, 23.0 +/- 2.5 days, and 19.4 +/- 2.1 days respectively, P0.001). Long survival (or= 60 days) rate in the rats treated with IL-12-IL-2 fusion gene on the first and third day was 30%. The serum mIL-12 and hIL-2 levels in these rats remained high on the 60th day after treatment. The pathological study showed that the number of infiltrating lymphocytes in liver tumor tissues was increased in the IL-12-IL-2 fusion gene treatment group.The retroviral packaging cell line injected intrasplenically encoding mIL-12 and hIL-2 fusion gene inhibits the growth of hepatocellular carcinoma significantly in rats. The therapeutical efficacy of early administration is superior to that of late one.

We have previously identified a human receptor protein tyrosine phosphatase of the MAM domain family, termed PCP-2, in human pancreatic adenocarcinoma cells and found that this protein was colocalized with beta-catenin and E-cadherin at cell junctions [Wang, H.-Y., et al. (1996) Oncogene 12, 2555-2562]. Its intracellular part consists of two tandem phosphatase domains and a relatively large juxtamembrane region that is homologous to the conserved intracellular domain of cadherins, suggesting a role in the regulation of cell adhesion. This study reports that PCP-2 was endogenously expressed at the cell surface and upregulated with increased cell density. An in vivo binding assay revealed that PCP-2 could directly interact with beta-catenin through a region in the juxtamembrane domain. Tyrosine phosphorylation of beta-catenin by EGF or active SrcY527F did not disrupt the formation of the PCP-2-beta-catenin complex, while PCP-2 in this complex could cause a significant reduction in the phosphorylation level in beta-catenin. Finally, we showed that PCP-2 was a negative regulator for cell migration. In conclusion, interaction of PCP-2 with its substrate beta-catenin is involved in the process of cell-cell contact.

AIM: To investigate cytpchrome P4502E1 (CYP2E1) gene expression in occurrence and progression of hepatocellular carcinoma (HCC). METHODS: The human liver arrayed library was spotted onto the nylon membranes to make cDNA array. Hybridization of cDNA array was performed with labeled probes synthesized from RNA isolated from HCC and adjacent liver tissues. Sprague-Dawley rats were administrated diethylnitrosamine (DENA) to induce HCC. CYP2E1 expression was detected by the method of RT-PCR and Northern blot analysis. RESULTS: CYP2E1 was found by cDNA array hybridization to express differently between HCC and liver tissues. CYP2E1 only expressed in liver, but did not express in HCC tissues and expressed lowly in cirrhotic tissues. In the progression of cirrhosis and HCC, the expression level of CYP2E1 was gradually decreased and hardly detected until the late stage of HCC. CONCLUSION: Using arrayed library to make cDNA arrays is an effective method to find differential expression genes. CYP2E1 is a unique gene expressing in liver but did not express in HCC. CYP2E1 expression descended along with the initiation and progression of HCC, which is noteworthy further investigations in its significance in the development of HCC.

AIM: To investigate the expression of p28/gankyrin gene and its role in the carcinogenetic process of human hepatocellular carcinoma (HCC). METHODS: 64 specimens of HCC and para-carcinoma tissues, 22 specimens of non-tumor liver tissues (7 normal, 15 cirrhosis), 10 specimens of normal human tissues and 5 hepatoma cell lines were studied for the expression of p28/gankyrin by Northern blot. The expression of p28/gankyrin protein was detected immunohistochemically by using the specific polyclonal antibody. RESULTS: Northern blot analysis indicated that the expression of p28/gankyrin mRNA was intensively distributed in brain and heart, weakly in lung, spleen and muscle, undetectable in digestive system including liver, pancreas, stomach, small and large intestines. p28/gankyrin mRNA was absent in normal liver, weakly detected in liver cirrhosis and in 18 of 64 para-carcinoma liver tissues. In contrast, the expression of p28/gankyrin mRNA was intensively detected in all 5 hepatoma cell lines tested, markedly increased in 57 of 64 and moderately increased in 5 of 64 HCC samples. In comparison with liver cirrhosis and para-carcinoma liver tissues, the average expression of p28/gankyrin mRNA in HCC was increased 3.6- (2.901 ± 0.507 vs 0.805 ± 0.252, P < 0.05) and 5.2-fold (2.901 ± 0.507 vs 0.557 ± 0.203, P < 0.01), respectively. In addition, p28/gankyrin mRNA expression level was higher in HCC with portal vein tumor thrombus and microscopic hepatic vein involvement (P = 0.021 and P = 0.047, respectively). The overexpression of p28/gankyrin protein in HCC was targeted in hepatic tumor cells, not in bile duct cells and other interstitial cells. CONCLUSION: Overexpression of p28/gankyrin in HCC plays an important role and contributes to the metastasis potential in the process of carcinogenesis. p28/gankyrin may become a specific biological tissue marker for the pathological diagnosis of HCC.