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Clock genes play a pivotal role in the rhythm maturation of bony fish ovaries. To explore the role of clock genes in the rhythmic ovarian development and maturation of tongue sole (Cynoglossus semilaevis), we used real-time fluorescent quantitative PCR technique to analyze the expression profiles of circadian locomotor output cycles Kaput 1a (Clock1a), brain and muscle Arntlike 1a (Bmal1a), Cryptochrome 1a (Cry1a), Cryptochrome 2 (Cry2), and Period 2 (Per2) at the ovarian stages Ⅱ, Ⅲ, Ⅳ, Ⅴ, and Ⅵ.The coding DNA sequence (CDS) of five clock genes were cloned and phylogenetically analyzed. We found that the CDS sequence length of Clock1a was 1 620 bp and encoded 539 amino acids, with the encoded amino acid sequence of Clock1a having a predicted molecular weight of 81.9 kDa. Clock1a has the functional domain PASD1 (consisting of 64 amino acids) and PAS11 (consisting of 103 amino acids). The CDS sequence length of Bmal1a was 1 881 bp and encoded 626 amino acids, with the encoded amino acid sequence of Bmal1a having a predicted molecular weight of 68.9 kDa. In the Bmal1a sequence, the functional domain PASD3 is composed of 63 amino acids, and the functional domain PAS11 is composed of 103 amino acids. The CDS sequence length of Cry1a was 1 896 bp and encoded 631 amino acids, with the encoded amino acid sequence of Cry1a having a predicted molecular weight of 71.4 kDa. Its functional domain FAD7 was composed of 199 amino acids. The CDS sequence length of Cry2 was 2 007 bp and encoded 669 amino acids, with the encoded amino acid sequence of Cry2 having a predicted molecular weight of 76.0 kDa. The Cry2 sequence contains a functional domain FAD7 consisting of 199 amino acids. The CDS sequence length of Per2 was 4 248 bp, encoding 1 415 amino acids, and the encoded amino acid sequence of Per2 has a predicted molecular weight of 154.0 kDa. In the Per2 sequence, the functional domain PeriodC consists of 295 amino acids and a functional domain PAS11 consists of 102 amino acids.The neighbor-joining method was used to analyze the Clock1a, Bmal1a, Cry1a, Cry2, and Per2 phylogenetic relationships between C. semilaevis and other bony fish, amphibians, birds, and mammals. The homology of Clock1a, Bmal1a, Cry1a, Cry2, and Per2 with other bony fish was 60%–79%, 94%–100%, 85%–91%, 84%–94%, and 70%–84%, respectively. Therefore, we believe that these five amino acid sequences show strong conserved property. In addition, the homology of Cry1a and Cry2 was 62%, indicating that Cry1a and Cry2 evolved differently during the evolution of C. semilaevis. In the constructed phylogenetic tree, Clock1a, Bmal1a, Cry1a, Cry2, and Per2 of C. semilaevis were clustered together with other bony fishes, indicating a close relationship between C. semilaevis and other bony fish in the evolutionary process. Moreover, the homology of Clock1a between C. semilaevis and mammals, birds, and amphibians is low, indicating that there are evolutionary differences in the evolutionary process of Clock1a. The high homology of Bmal1a, Cry1a, Cry2, and Per2 with mammals, birds, and amphibians suggests that these four clock genes were strongly conserved during the evolution of C. semilaevis.In the present study, we found that the expression levels of five clock genes were high in stagesⅡ and Ⅲ, which were equivalent to the non-reproductive season (P < 0.05). However, the expression levels of five clock genes were low in stages Ⅳ and Ⅴ, which were equivalent to the reproductive season (P < 0.05). Therefore, the expression profiles of Clock1a, Bmal1a, Cry1a, Cry2, and Per2 in the ovaries of C. semilaevis also have seasonal characteristics. The ovarian development and maturation of C. semilaevis goes through stages Ⅱ, Ⅲ, Ⅳ, Ⅴ, and Ⅵ, and then reaches stage Ⅱ again and starts a new reproductive cycle. The variation patterns of seasonal factors, such as light and temperature, in fish ovaries were consistent with the annual expression patterns of the five clock genes in this study. Therefore, it can be considered that the expression of clock genes in C. semilaevis has an annual cycle. The findings presented in this study can enrich the theory of ovarian development and maturation of C. semilaevis and provide a theoretical basis for improving breeding technology and seedling efficiency.

The whitespotted conger (Conger myriaster) is an important fish species with high economic value in China and Japan owing to its high nutritional content. It occupies an important position in marine-eel export in China. C. myriaster has the potential to be artificially cultured and has promising economic prospects. Thus, there is growing international interest in C. myriaster research. Indeed, to meet the demands of domestic and foreign markets, artificial culturing of C. myriaster is urgently required. However, at present, there is no report on the research of C. myriaster reared in industrial recirculating aquaculture systems. Feeding frequency has been reported to affect feed intake and growth performance in a number of fish species. Feed management in terms of optimization of feeding frequency has emerged as one of the crucial areas of research in the field of aquaculture. Overfeeding and leftover food disrupts the water quality, while inadequate food supply has direct impact on production cost. Traditional flow through systems uses large volumes of water, and intensified aquaculture has drawbacks, such as large amount of discharged waste from effluent water. Among the excreted nutrients, nitrogen and phosphorus constitute the largest proportion. Sustainability with low environmental impacts is one of the concerns when developing aquaculture systems. Based on the concepts of reduced water consumption and efficient water management, recirculating aquaculture systems offer the possibility of creating a sustainable future for fish production. Thus, a 70-day experiment was conducted to investigate the effects of different feeding frequencies and water quality on the growth performance and physiological and biochemical indices of C. myriaster [body weight: (150.64±5.43) g] reared in industrial recirculating aquaculture systems; physiological and ecological methods were used in this study. The experiment comprised two feeding frequency groups, feeding once (group T1) and twice (group T2) per day, and each treatment group had three replicates. The average body weight of the fish in each culture pond and the water quality indices of the culture system were measured and analyzed every 14 d. At the end of the experiment, the blood, liver, and intestine samples of C. myriaster were collected to determine and analyze growth and physiological indices. The results indicated that (1) the growth performance of group T2 was better than that of group T1. The final body weight, final body length, weight gain, and specific growth rate of group T2 was significantly higher than those of group T1 (P < 0.05). The feed coefficient of group T1 was significantly higher than that of group T2 (P < 0.05). However, no significant differences were found in the final body height, liver body ratio, visceral body ratio, condition factor, or survival rate at different feeding frequencies (P > 0.05). (2) The serum malondialdehyde of group T1 was significantly higher than that of group T2 (P < 0.05). The hepatic alanine aminotransferase and hepatic aspartate transaminase of group T1 was significantly higher than those of group T2 (P < 0.05). The intestinal trypsin of group T2 was significantly higher than that of group T1 (P < 0.05). There were no significant differences in the levels of serum superoxide dismutase, serum catalase, intestinal amylase, or intestinal lipase between groups T1 and T2 (P > 0.05). (3) In the 24 h changes of ammonia nitrogen (NH4+-N) and nitrite nitrogen (NO2–-N), the change patterns of the two indices in outlet of the T1 group were similar to those of the outlet of the bio-filter; the two indices of the outlet water in T1 and T2 groups were one (24:00) and two peaks (20:00 and 08:00) respectively; the difference in NH4+-N in the outlet water of the two test groups within 24 h was significant (P < 0.05); at 04:00 and 08:00, the NO2–-N content of the outlet water of the biofilter was significantly lower than that of the other two test groups (P < 0.05); the 24 h changes range of NH4+-N and NO2–-N in T2 group were smaller than those in T1 group. (4) During the whole test period, each water quality index was maintained within the safe breeding range, and no significant differences were found in NH4+-N, NO2–-N, chemical oxygen demand, and active phosphate concentration between T1 and T2 groups (P > 0.05); at the end of the test, the total nitrogen and total phosphorus of T2 group were significantly higher than those of T1 group (P < 0.05); the biofilter has different treatment effects on various water quality indices, among which the treatment effect on NH4+-N is the most efficient, up to 54.23%. Based on the above results, the feeding frequency of twice per day was better than that of once per day in the industrial recirculating aquaculture mode for C. myriaster (body weight: 150~250 g). Without affecting the survival rate and condition factor, the twice per day group can save costs and improve economic benefits. However, due to the limitations of the test conditions, we did not explore more times of feeding, and further tests are needed in the future. The predominance, which was the high efficiency of water qualities treatment and optimal effect of culture, was demonstrated obviously in closed recirculating aquaculture systems. This study aims to provide a theoretical reference for the appropriate feeding strategy in the industrial recirculating aquaculture model of C. myriaster.

Moderate clothes-changing person re-identification is to find the same person from different camera scenes under the premise of considering the moderate change of clothes.The implementation of existing person re-identification methods is usually based on the assumption that the pedestrian’s clothing is invariant,so they rely on clothing-related features.Then,when the above assumptions are not valid,these methods are difficult to achieve the ideal recognition performance.Considering the important characteristic that pedestrian’s shape hardly change when the change of clothes is moderate,the moderate clothes-changing person re-identification is studied.Inspired by the binocular summation in biological vision system,a self-attention siamese network is proposed in this paper.Analogous to biological binocular information acquisition process,the network takes two types of images of the same pedestrian with different clothes as two branch inputs,and then achieves summation effect with siamese architecture.Subsequently,the contrastive learning and fusion learning of multiple features are carried out to obtain the pedestrian feature representation with identity discrimination.Finally,empirical studies show that the proposed method achieves best performance at present on clothes-changing person re-identification benchmark.

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Purpose: To evaluate the accuracy of static computer-navigated implantation with surgical guides, based on a non-radiological method. Traditional measurements with a second cone-beam CT (CBCT) were applied to verify the accuracy., Methods: A total of thirty template-guided implantations were designed and performed on 15 resin models. Two paralleled bone-level implants were planned in the edentulous space of each model, between which the distance was 4 mm. Postoperative implant positions were detected with both CAD/CAM-based measurements applying an intraoral scanner (3Shape TRIOS) and traditional ways via CBCT. Both methods were conducted with a CAD quality-control, reverse engineering software, Geomagic Studio 2013, comparing the positions with the virtual ones. Statistical analysis was processed with SPSS 23.0 software package., Results: Measurements using CBCT (control group) showed a trend toward greater deviations when the results were directly compared(P<0.05). In the CAD/CAM-based evaluation of the 30 samples, the mean deviation of the insertion axis from the planned implant axis was 1.134°. The mean deviations of the implant shoulders in the horizontal direction and at the implant apices were 0.447 mm and 0.557 mm, respectively. No significant difference was observed when measuring distance deviation with the two assessment ways., Conclusions: Compared with evaluation based on radiology, CAD/CAM based evaluation system is able to evaluate implant accuracy precisely, effectively reduce radiological exposure of patients, being suitable for clinical evaluation.

Purpose: Using impact test to study cushioning and energy absorption of sports mouthguard of different thickness, material and combination., Methods: According to the thickness, material and combination, the mouthguards were divided into 7 groups with 6 testing pieces in each group. A pendulum device was used to apply impact energy to the mouthguard and dentition model, and the magnitude of the impact energy was measured by changing the release height of the pendulum. The force sensor and the laser vibrometer were respectively connected to the data acquisition and signal analyzer to obtain the incident speed, rebound speed, maximum collision force, collision start time and collision end time of the hammer head. Collision time, energy absorption and energy absorption ratio of the mouthguard were calculated, which reflected different mouthguard cushioning and energy absorption characteristics. One-way analysis of variance and t test in SPSS 22.0 software package was used to analyze the difference between different groups., Results: No matter what type of mouthguard was worn by the dentition model, the impact force was significantly reduced than without mouthguards. At the same height, there was significant difference between the average maximum collision force and the average collision time among soft mouthguards with thicknesses of 2, 3, 4, and 5 mm(P<0.05); among which the average maximum collision force of the 5 mm was the smallest and the average collision time was the shortest. There was no significant difference in the average energy absorption and the average energy absorption ratio among these 4 groups. There was significant difference in the average maximum collision force, average collision time, average energy absorption and average energy absorption ratio between the two groups of mouthguards with the same thickness of splints and different materials(P<0.05). Among them, the average maximum impact force of the 4 mm soft mouthguard was smaller and the average collision time was longer., Conclusions: Wearing sports mouthguard can reduce the risk of tooth fracture during collision. Soft mouthguard has the best cushioning performance and it is positively related to thickness. Locally strengthening hard materials can enhance energy absorption and provide better protection.

Purpose: To investigate the effect of tooth movement at different time after repair of alveolar bone defects with tissue engineering bone constructed by rabbit bone marrow mesenchymal stem cells (BMSCs) and beta-tricalcium phosphate (β-TCP)., Methods: Alveolar bone defect (6 mm×4 mm×8 mm) was made on the right side of 40 New Zealand rabbits, which was filled with tissue engineering bone constructed by BMSCs and β-TCP as experimental sides. Tooth extraction on the other side was performed as control. The mandibular second molars in both sides were moved mesially 2, 4, 8 and 12 weeks after surgery. The specimens were taken 4 weeks after exertion. The distance of mandibular second molar moved was measured. The periodontal tissues were observed after H-E staining. TRAP staining was performed and osteoclasts were counted in the periodontal tissues on the pressure side. BMP-2 immunohistochemical staining was used to observe the average optical density of periodontal tissue on tension side. The results were analyzed with SAS 8.0 software package., Results: The distance of tooth movement, the amount of TRAP positive cells and the optical density of BMP-2 in the experimental side of the 2-week and 4-week groups were all significantly lower than those in the control side, while there was no significant difference between the 8-week and 12-week groups., Conclusions: Eight weeks after repair of alveolar bone defect with rabbit BMSCs and β-TCP is an appropriate time for orthodontic tooth movement.

Purpose: To explore periodontal responses on the compression side during early tooth movement into alveolar defect regenerated by bone marrow mesenchymal stem cells (BMSCs) and porous granulated beta-tricalcium phosphate（β-TCP） scaffolds., Methods: Thirty New Zealand rabbits were used to establish bilateral mandibular defective alveolar bone model by extracting the mandibular first molars and expanding the sockets. The right mandibular alveolar defects were filled with a construct of β-TCP scaffolds combined with BMSCs as experimental group. The left alveolar defects were repaired by β-TCP scaffolds alone as control group. Eight weeks later, 6 rabbits were sacrificed to evaluate osteogenesis effect. The other rabbits were loaded orthodontic force to move the bilateral second molars forward for 4 weeks. Six rabbits in each group were sacrificed at 1, 2, 3, and 4 week after orthodontic tooth movement (OTM). The distance of OTM was measured, and the status of periodontal tissues was observed by H-E staining. The number of osteoclasts on the compression side of tooth was counted by tartrate-resistant acid phosphatase histochemistry. The results were compared between groups using SPSS 19.0 software package., Results: After 8 weeks of bone grafting, the osteogenesis effect of the experimental group was better than the control group. The OTM distance in the experimental area was higher than that in the control area. At 2, 3 and 4 week of OTM, the number of osteoclasts in the experimental group was significantly higher than that in the control group., Conclusions: A tissue-engineered complex with β-TCP scaffolds and BMSCs could well repair the alveolar bone defect. When the adjacent tooth moved into regenerated area, the new periodontal tissue had an active response, promoting to accelerate tooth movement.

To clarify the effect of invasive alien plants on ecosystem functions, soil total carbon (TC), total carbon (TN), and total phosphorus (TP) contents were studied in the exotic invasive plant Spartina alterniflora marsh with different durations and the native species Cyperus malaccensis marsh in the Minjiang River estuary. The results showed that there were different variations of TC, TN and TP at 0-50 cm soil layer in the C. malaccensis marsh invaded by S. alterniflora, but the changes of TC and TN were similar and the change of the TP lagged behind them. Soil C/N increased continually with the increase of TC. Soil TP was a key factor affecting soil C/P and N/P during the S. alterniflora invasion, and the change of C/P and N/P followed a similar trend. The changes of TC, TN and TP were influenced by abiotic factors such as soil salinity, bulk density, soil water content and clay composition. C, N and P stoichiometric ratios were influenced mainly by soil salinity and grain composition. C/N and C/P were the good indicators for soil carbon enrichment ability in S. alterniflora marsh. S. alterniflora invasion induced the change of biomass and marsh habitats. Soil carbon, nitrogen, phosphorus contents, and their ecological stoichiometric ratios changed significantly in the C. malaccensis marsh invaded by S. alterniflora and showed diffe-rent change characteristics with different invasion duration.

Objective: To explore the diagnostic accuracy of optical coherence tomography (OCT) and intravascular ultrasound (IVUS) in the detection of ex vivo coronary plaques with different compositions compared with histology results., Methods: OCT and IVUS were performed in 15 autopsied heart specimens and the isolated coronary artery was assessed by routine histological processing thereafter. Coronary plaques were classified into 3 types (lipid-rich plaque, calcified plaque and fibrous plaque) according to standard criteria respectively. Sensitivity and specificity for detection of different types of plaque by OCT and IVUS were calculated according histology results., Results: Seventy seven coronary plaques were analyzed. OCT demonstrated a sensitivity and specificity of 69% and 88% for lipid-rich plaque, 93% and 92% for calcified plaque, 88% and 98% for fibrous plaque. IVUS demonstrated a sensitivity and specificity of 61% and 92%, 98% and 97%, 68% and 90% respectively. The agreement between OCT and IVUS in assessment of coronary plaque was 0.831 (Kappa = 0.72, P < 0.01)., Conclusions: Both OCT and IVUS correctly detected ex vivo coronary plaques and there was a good agreement in assessment of coronary plaques between OCT and IVUS. OCT is superior to IVUS in assessment of fibrous plaque and is similar as IVUS in assessment of calcified plaque.

Objective: To define the role of interferon-γ on radiotherapy of lung cancer and explore a new way to clinical treatment., Methods: A549 cells were exposed to γ ray with or without IFN-γ co-treatment. MTT assay was performed to evaluate cell viability. Western blot was used to observe the expression of P53 protein., Results: The results showed that co-treatment of IFN-γ decreased the cell viability significantly compared with the γ ray irradiation group (71.4% ± 2.1% vs 44.1% ± 3.1%, n = 7, P < 0.01). In addition, the expression of P53 protein also increased significantly after co-treatment (P < 0.01); Furthermore, the cell cycle was changed obviously in co-treatment group compared with γ ray irradiation group, S phase increased (12.9% vs 20.9%, n = 5, P < 0.05) and also blocked the G2/M phase (28.8% vs 38.9%, n = 5, P < 0.05)., Conclusions: The results suggested that γ ray irradiation combined with IFN-γ can increase the efficiency of radiotherapy on A549 cells and there is much broad prospect in the clinical treatment of lung cancer.

Objective: To explore the clinical characteristics, diagnosis and surgical treatment of adult congenital bronchoesophageal fistula., Methods: Eleven cases of adult congenital bronchoesophageal fistula that were diagnosed and surgically treated between May 1990 and August 2010 had been reviewed. There were 7 male and 4 female patients, ranging in age from 28 to 66 years (mean 48.7 years). The chief clinical presentation included coughing and sputum in 10 cases, recurrent bouts of coughing after drinking liquid food in 6 cases, hemoptysis in 6 cases, low fever in 4 cases, chest pain in 3 cases. The duration of symptoms before diagnosis ranged from 5 to 36 years (mean 16.8 years). The diagnosis of bronchoesophageal fistula was confirmed most by esophagography. Associated diseased lung was resected in all patients (lobectomy in 10 cases and pneumonectomy in 1 case). The operation included right thoracotomy in 7 cases and left thoracotomy in 4 cases. The fistula was completely resected in 10 cases. The tract was simply divided and the end was sutured in 1 case., Results: The postoperative course was uneventful in 10 patients who were discharged from hospital 10 to 18 d after operation. One patient suffered from esophageal fistula and received second operation. Regular follow-up was conducted on all 11 patients, proving that 3-year survival rate was 11/11 and 5-year survival rate was 9/11., Conclusion: Persistence of congenital bronchoesophageal fistula into adulthood is rare. The main symptom is nonspecific coughing and bouts of coughing after drinking liquid food. The most useful diagnostic method is the esophagography. Even though it is benign disease, life-threatening complications might occur and it must be treated surgically as soon as the diagnosis is established.

Objective: To investigate the histologic effect of botulinum toxin type A (Botox A) injection on myocutaneous flap expansion in minipigs model., Methods: Seven minipigs were included in this study. Two symmetric tattoo area, 10 cm x 6 cm in size, were selected on the bilateral flank of the pigs. The Botox A was injected into one tattoo area randomly, 4 U every point, 2 cm apart, with a total dose of 96 U. The same dose of sterile normal saline (0.9%) was injected in the same fashion on the opposite side as control. 3 days after injection , two 200 ml expanders were inserted beneath the cutaneous muscle at the tattoo area. After complete expansion of 200 ml, the specimens were drawn from both groups symmetrically and were stained by means of HE and Masson. The histologic changes of myocutaneous flap were compared. Thickness of each layer in myocutaneous flap was measured in histological section., Results: The thickness of cutaneous muscle, capsule, dermis were (275.74 +/- 28.93) microm, (468.03 +/- 34.28) microm, (1990.79 +/- 102.10) microm in Botox group, and (409.13 +/- 44.63) microm, (626.55 +/- 44.05) microm, (2508.44 +/- 70.71) microm in saline group, respectively, show a significant difference between the two groups (P < 0.01). The Masson stained slice showed that collagen average gray of capsule in Botox group was 185.38 +/- 9.56, which was significantly higher than that in the saline group (120.77 +/- 10.31, P < 0.01). Light microscope (HE stained sections ) showed that muscle in Botox group was significantly atrophy and cross-section of muscle fiber decreased. The muscle fiber in saline group was generally normal. It was observed through transmission electron microscope that the light and dark band of muscle cell became fuzzy and the Z line bending in Botox group. The light and dark band in saline group arranged neatly, the Z line was clear., Conclusions: Application of Botox A in myocutaneous flap expansion can make the muscle atrophy and reduce the content of collagen in capsule layer, making the myocutaneous flap thinner which is suitable for reconstruction in face and neck.

Objective: To explore the value of partial atrium or large blood vessel resection for the treatment of locally advanced lung cancer., Methods: Thirty-five patients with locally advanced lung cancer (T(4)N(0)-N(2)M(0)) underwent lobectomy or pneumonectomy combined with intrapericardial vascular management or partial resection of the atrium. Of the 35 patients , 15 underwent left pneumonectomy combined with partial resection of the left atrium, 3 had pneumonectomy and partial resection of pulmonary artery trunk, 11 received right pneumonectomy and partial resection of the left atrium, 3 had middle and lower lobectomies and partial resection of the left atrium, and 3 underwent right upper lobectomy, partial resection of the superior vena cava and replacement of artificial blood vessel., Results: No death occurred in the 35 patients. Postoperative arrhythmia occurred in 4 cases and respiratory failure in 2 cases. The 1, 2, 3 and 4 year survival rates of the patients were 79.2% (19/24), 53.3% (8/15), 46.2% (6/13) and 36.4% (4/11), respectively. Pathologically, 27 patients had squamous carcinoma, 3 had adenocarcinoma, 3 had adenosquamous carcinoma and 2 had large cell carcinoma. In TNM staging, 6 were in T(4)N(0)M(0), 11 in T(4)N(1)M(0) and 18 in T(4)N(2)M(0)., Conclusion: Pneumonectomy or lobectomy combined with intrapericardial vascular management or partial resection of the atrium can enhance the possibility of radical resection of locally advanced lung cancer and increase the long term survival rate.

Objective: To investigate the method of early surgical treatment of bronchopleural fistula after pneumonectomy., Methods: Twelve patients (9 males and 3 females with a mean age of 58.6-/+5.7 years) with bronchopleural fistula after pneumonectomy received a reoperation within 72 h after a definite diagnosis. Empyema was found in none of the 12 cases. Fistula occurred within 4 to 17 days (8 days in average) after the operation. The fistula of the residual main bronchus was resected, and the thoracic cavity was asepticized by flushing., Results: Ten patients were discharged with complete healing. One patient was discharged following open drainage with daily change of the wound dress. One patient died due to multiple organ failure. The hospital stay of the patients ranged from 18 to 49 days (31 days in average) after the reoperation., Conclusion: Bronchopleural fistula after pneumonectomy, in case that empyema and multiple organ failure do not occur, can be healed by closing the fistula with the stapling device in early stage. Flushing the thoracic cavity is also necessary after the reoperation.

Objective: To investigate the relationship between the vasoactive substances including endothelin-1 (ET-1), calcitonin gene-related peptide (CGRP), and nitric oxide (NO) in myocardium and the cardiac functions in chronic renal ischemia rats., Methods: Male Wistar rats weighting 180-200 g were randomly divided into 2 groups: operation group (n=30) and sham operation group (n=10). A ligation of abdominal aorta between right and left renal artery was made by silk suture in operation group and the necrosis degree of aorta was about 50%. Aorta was not ligated in sham operation group. Sixteen weeks after operation, invasive measurement of blood pressure and cardiac function were performed, and content of ET-1, CGRP, and NO in myocardium were determined., Results: Compared with sham operation group, blood pressure significantly increased in the operation group after ligation, along with decreased cardiac systolic and diastolic function, increased left ventricular mass index. After 16 weeks, compared with sham operation group, the content of ET-1 in cardiac tissue were significantly elevated in operation group [(361.0+/-118.7) vs. (503.4+/-139.6 ) pg/ml, P<0.01), along with significantly decreased CGRP content [(74.4+/-24.8) vs. (45.4+/-15.1) pg/ml, P<0.01). The content of ET-1 in cardiac tissue was negatively correlated with the maximum pressure rise rate of left ventricular r=-0.37, P<0.05)., Conclusions: Chronic kidney ischemia caused by abdominal aorta ligation may result in the increase of ET-1 content in cardiac tissue and decrease of CGRP decreased. The content of ET-1 in cardiac tissue is also negatively correlated with left ventricular systolic function.

A dominant wheat stripe rust resistance gene, temporarily designated as YrY201, was identified in an accession Y201 of Aegilops tauschii. By bulk segregation analysis, three microsatellite markers Xgwm273b, Xgwm37 and Wmc14 were found to be linked to YrY201 with genetic distance of 11.5, 5.8 and 10.9 cM , respectively. According to the locations of the linked markers, the resistance gene was located on chromosome 7DL. Based on the chromosomal location and the resistance pattern of the gene, we proposed that YrY201 was a novel stripe rust resistance gene, and could be selected by marker-assisted selection.

To constructed the recombinant human anti-rabies virus ScdsFv, cys sites were introduced into framework region (FR) of VH and VL genes which were amplified from human anti-rabies virus ScFv respectively using genetic point mutation technology. Cloned the ScdsFv gene into expression vector pET22b (+) and transformed into E. coli BL21 (DE3). The target protein was expressed by inducing with IPTG. Followed by renaturation in vitro and purified by Ni-NTA. The binding activity of ScdsFv was identified by Fluorescent antibody test (FAT) and ELISA. Results showed that recombinant ScdsFv were expressed at high level. Purity of the protein > 90% after purified by Ni-NTA and renaturaton in vitro. FAT and ELISA results demonstrated that ScdsFv could binding antigen specificity and was more stable than ScFv. Recombinant ScdsFv provided experiment materials for further functional study.

Objective: To study the effect of GMCSF-absence on the rate of wound healing and neovascularization during wound repair., Methods: 30 wild type (WT) mice and 30 GMCSF- absence mice (GMCSF-KO) were obtained. They were received full thickness skin wound (0.8 cm x 0.8 cm) in each side of midline after deeply anesthesia. In the different post-injury time points, the wound sites were digitally photographed to calculate the percentage of wound closure by using computer image analyses software. The wound specimens were also obtained dynamically for immunohistological analysis of CD31 at wound site., Results: The analysis of the wound closure showed that wound healing in GMCSF-KO mice was delayed significantly comparing with that in WT mice from the day 3 post-wounding. At days 7 and 10 after wounding significantly more numbers of blood vessels were formed in the WT controls compared to the GMCSF-KO mice., Conclusions: GMCSF-absence inhibits neovascularization during wound repair and leads to the delay of wound healing.

Aim: To express the anti-rabies MBP-ScdsFv fusion protein in E.coli and identify it's activity assay., Methods: A full length of ScdsFv gene was Cloned into prokaryotic vector pMAL-p2x by recombinant DNA, Then the recombinant plasmid pMAL-ScdsFv was transformed to E.coli ER2566 for expression and induced by IPTG. The fusion protein was digested by Factor Xa, purified by Ni(+) and amylose resin affinity chromatography and Identified by SDS-PAGE and Western blot. The binding activity of the protein was determined by ELISA., Results: Recombinant plasmid pMAL-ScdsFv was successfully constructed. The anti-rabies ScdsFv protein which was digested by Factor Xa and purified by Ni-NTA and amylose resin affinity chromatography had good binding activity and high stability., Conclusion: The anti-rabies ScdsFv protein is of good binding activity and high stability, and it can be used for the immune therapy.

In order to explore the resistance and the staphylococcal chromosome cassette mec (SCCmec) types of Methicillin-resistant S. aureus (MRSA) in the area of Haikou, 686 strains of MRSA had been distinguished from 1174 strains of S. aureus using PBP2a testing. The resistance to the seven deputies of seven kinds antibiotics which in common use in clinic, including Oxacillin, Vancomycin, Doxycyclin, Amikacin, Erythromycin, Chloramphenicol, Ciprofloxacin, and SCCmec type of 58 strains had been tested using the K-B Agar diffuse, E-test and multiplex PCR strategy, and seven kinds of new SCCmec types were found in 17 strains. Their specialties of structure are: type-new3 possess four loci of A, F, H, M; New4 possess three loci of F, H, M; New5 possess three loci of D, B, M; New6 possess three loci of A, B, M; New7 possess four loci of H, E, C, M; New8 possess two loci of A, M; New9 possess three loci of A, C, M. All of them are different from the types reported. The strains carrying new SCC mec types are different from that carrying old SCC mec types in the epidemical distribution and resistance to the antibiotics: they were mostly isolated from the out-patients and have high level and wider range of resistance to antibiotics and deserve to pay more attention.

In order to explore the resistance, epidemical distribution and the staphylococcal chromosome cassette mec (SCCmec) types of Methicillin-resistant S. aureus (MRSA) in the area of Haikou, the resistance and SCCmec type of MRSA isolated from clinical sample has been tested using the K-B Agar diffuse, E-test and multiplex PCR strategy, resulting that two kinds of new SCC mec types were found in them. Type-newl possess three loci of B, M, F. Locus M, which locate in mecA, is the indicator of mecA, and also used as the positive control. Locus B, which locate in upstream of mec A, is specific for type- II and F, which locate in downstream of mec A, is specific for type- iU ; and so, type-newl was combined from the type- II and - I . Type-new2 possess five loci of A, B, M, F, H. Locus A, which is locate in upstream of mecA, is specific for type- I and H, which is locate in downstream of mecA, is also specific for type- 1 ; and so, type-new2 was combined from the type- I , - II and - I . The strains carrying new SCC mec types are different from that carrying old SCC mec types in the epidemical distribution and resistance to the antibiotics. They can be usually isolated from the out-patients and have high level and wider range of resistance to antibiotics. It must be paid more attention to them.

The choice of specific promoters used within a transgene construct is a vital strategy to achieve the transgene regulation in the temporal, spatial and measurable manner. The strategy has been widely used in diverse aspects of plant gene engineering, such as quality improvement, resistance breeding and bioreactor. In this paper, we describe the structure feature, classification and research method of the specific promoter and its application progresses in plant gene engineering.

Under induction with 41 degrees C, pBV220-stxb/DH5alpha expressed the recombinant protein Shiga Toxin B Subunit (StxB), which was purified by centrifugation, salting out and ion exchange chromatography. As a target, the purified-protein-coated ELISA plate was used to screen phages able to bind onto it from a random 12-mer peptide library. After 4 rounds of affinity screening, a group of clones were isolated from the peptide library. ELISA assay detected their binding activity with the target. 27 clones showed the specific binding activity. The peptide sequences of these positive phage clones were analyzed. 16 of them had the same sequence named A6, 2 clones had the A9 sequence, and 3 clones had the A3 sequence. To evaluate the neutralization effect of A6 phage, animal test was carried out. The Shiga Toxin was incubated with A6 phage clone, then used to attack the Balb/C mice. As a control, the toxin was incubated with negative phage. In the control group, no mice survived. Comparing with it, the survival rate of the mice in neutralization group could reach to a level of 33.3%. It showed that the toxicity of Shiga Toxin was partly inhibited. The A6 peptide could be developed as an inhibitor of Shiga Toxin to cure the diseases caused by Shiga Toxin.

Leptospira interrogans (L. interrogans) genomic DNA was used as template to amplify the full-length gene for ribosomal protein L11 methyltransferase (liPrmA) by PCR. The pET22b-/liprmA expression plasmid was successfully constructed in Escherichia coli (E.coli.) strain TOP10 and confirmed by restriction enzyme digest and sequencing. Through optimizing expression of the recombinant liPrmA-6xHis fusion protein in expression host E. coli. BL21, the yield of soluble target protein reached 40 mg (liter culture)-1. The LiPrmA was purified to apparent homogeneity in a single step using Ni-NTA His Bind chromatography. Amino acid homologous analysis showed that liPrmA shared significant identity with other prokaryotic PrmA and eukaryotic putative PrmA in the catalytic region including AdoMet binding domain. Methylation activity experiments showed purified liPrmA was able to catalyze the ribosomal protein L11 of L. interrogans methylated under the presence of S-adenosyl-methionine (AdoMet).

With culture-independent approach, microbial total DNA was directly extracted from Pachychalina sp. Using the total microbial DNA as template, archaeal 16S rDNAs were amplified by PCR with universal primers. Amplified products were cloned into T-vector and secondarily amplified by PCR. Then the secondarily amplified products were purified to be further characterized by termed ARDRA (amplified rDNA restriction analysis). According to the enzyme restriction mapping, the apparent difference among them were disclosed. Furthermore eight archaeal cloned partial sequences were acquired and built up a phylogenetic tree. In the phylogenetic tree, the eight archaea belonged to Methanogenium organophilum and Methanoplanus petrolearius, but the 16S rDNAs similarities among them and those archaea registered in RDP Database didn't excess to 90%. It means that they maybe represent some novel archaeal groups.

Objective: To explore the methodology and operative essentials of laparoscopic cholecystectomy with remote Zeus surgical robotic system., Methods: Based on strict training and successful experiment in animal model of swine, laparoscopic cholecystectomy using Zeus robotic system was performed on 16 patients with biliary diseases, including choledocholithiasis, cholelithiasis, polyposis of gallbladder, and chronic cholecystitis, 10 males and 16 females, aged 33 (14 approximately 27), 26 April to 31 August 2004. The general data, preoperative preparation time, operation time, amount of bleeding, complications, and hospitalization time were analyzed., Results: All operations were performed without event. Along with the accumulation of experience the preoperative preparation time was shortened from 90 min to 30 min with an average of 41.7 min, and the operation time from 120 min to 30 min with an average of 64.4 min. The average amount of bleeding was 27.7 ml, and the average postoperative hospitalization time was 2.4 d. A telephone follow-up 30 days after operation showed no abnormality., Conclusion: Laparoscopic cholecystectomy with Zeus surgical robotic system is feasible and reliable with the advantages of clearer images in the field of operation, more precise handling, and remote surgery or education.

Aim: To clone the canine IL-18 cDNA and to explore the immunological effectiveness of canine IL-18 as an adjuvant of genetic vaccine., Methods: Canine leukocytes were separated from peripheral blood stimulated with ConA for 5-12 h. The full length cDNA of canine IL-18 was amplified by RT-PCR using leukocyte mRNA as a template. IL-18 cDNA was cloned into pMD18-T. Sequencing result showed that the full length cDNA was 582 bp, encoding 193 amino acids, which was identical with that published. The eukaryotic expression vector pIIL18 was constructed. Dogs were inoculated in mixture form of pIGneo and pIIL18., Results: Canine IL-18 cDNA was successfully cloned and eukaryotic expression vector pIIL18 was constructed. The immunological assay result showed that the anti-rabies virus-specific antibody level of the group immunized with mixture of pIIL18 and pIGneo was obviously lower than that of the group immunized with pIGneo alone, but the level of cellular immunity of the former was higher than the latter., Conclusion: Canine IL-18 can enhance cellular immunity but at the same time suppress humoral immunity, which lays the foundation for IL-18 as an adjuvant of genetic vaccine.

Aim: To study the influence of FasL overexpression in transgenic mice on the Sertoli cell immune response to testicular infection., Methods: Ureaplasma urealyticum (UU) was directly injected into bladders of FasL transgenic mice and wild-type mice respectively, to mimic an ascending infection pathway. At week 1, 2 and 3 after injection, respectively, the mice were put to death to observe the pathological alterations in testis section. And at the same time the differences of FasL, TGF-beta, IL-1alpha and IL-6 expressions on Sertoli cells were compared by immunohistochemical staining between wild mice and transgenic mice before and after infection, respectively. The high-purified Sertoli cells were isolated from the testis tissue of wild-type mice, comparing apoptotic capability of Fas(+) Jurkat cells mediated by FasL(+) Sertoli cells of wild control and wild UU-infected groups., Results: The pathological changes of testis tissue from transgenic mice were more serious as compared with wild-type mice and the model of cytokines secreted by sertoli cells was distinctive between the two kinds of mice. The UU-infected Sertoli cells increased Fas(+) Jurkat cell apoptosis., Conclusion: FasL overexpression can influence the cytokine's secretion in the process of anti-infection immunity and further affects the immune balance of testis locality. Not all FasL over expression is benefit to body's anti-infection immune response.