Objective To investigate the application value of transcystic choledochoscopy combined with holmium laser for common bile duct stones . Methods Clinical data of 16 patients with common bile duct stones who underwent transcystic choledochoscopy and holmium laser cholelithotripsy between March 2012 and December 2013 in this hospital were retrospectively analyzed.During holmium laser cholelithotripsy (1.0 J/10 Hz, 400 μm fiber), the stones were fragmented under direct vision of choledochoscopy and were extracted by using water flushing or a stone basket . Results The stones were successfully removed in 15 patients, including 6 cases of laparoscopic operation and 9 cases of open operation .The operation time was (132 ±27) min for open surgery and (156 ±33) min for laparoscopic surgery .The stone fragment and removal time was (30.2 ±8.5) min for open surgery and (45.6 ±10.4) min for laparoscopic surgery .The hospital stay was (9.7 ±1.4) d.There was 1 case of failed cholelithotripsy due to obstructed water flow and blurry vision .Follow-up examinations in 14 cases for 2-24 months found no recurrence of stones or biliary strictures. Conclusion Transcystic choledochoscopy and holmium laser cholelithotripsy is intuitive , accurate, and effective, being a safe and reliable alternative for choledocholithiasis .

Pancreatic cancer has a very poor prognosis, with a five-year survival less than 5%. Nowadays, to improve the treat-ment efficacy of pancreatic cancer has been a highlight for medical researchers worldwide. Increasing knowledge about the molecular pathogenesis of the disease has shown that genic mutations, such as oncogene Kras2, and anti-oncogenes Cdkn2a, and TP53, are hall-marks of pancreatic cancer. Therefore, it is vital to deeply understand the pathogenetic mechanism of pancreatic cancer and find new therapeutic strategies to cover it. In this review, we summarized the recent advances in the signaling pathways of pancreatic cancer.

Objective　To explore the phenomenon of neural apoptosis after chronic compressive spinal cord injury. 　Methods　A newly designed chronic progressive compressive spinal cord injury model of rat was adopted in this study. Forty-five Wister rats were divided into mild, moderate, and severe group according to the compressive degree of the spinal cord. The Feulgen stain and TUNEL were used to investigate the apoptosis and its characteristics in different kinds of neural cells. 　 Results　Apoptosis index in moderate injury group was the highest. Apoptotic cells largely located in ventral, lateral and dorsal column of the white matter. Most of them were oligodendrocytes. Positive neuron occasionally presented in laminae Ⅲ～Ⅸ part of the sections. Most of them located in dorsal gray horn. 　Conclusions　 Apoptosis is an important event in secondary pathophysiological process of chronic progressive compressive spinal cord injury. Apoptosis is one of the reasons for neural cell death. The apoptosis of oligodendrocytos may contribute to myelin sheath disruption of white matter.

Objective To investigate the effect of a selective inhibitor of COX-2 nimesulide on growth and apoptosis of human cholangiocarcinoma cell line QBC939 in vitro. Methods MTT assay was used to determine the influence of nimesulide on the proliferation of QBC939 cells, apoptosis of QBC939 was measured by transmission electron microscopy and flow cytometry.Expression of apoptosis related genes mRNA and bcl-2 ,bax, survivin were detected by RT-PCR and immunocytochemistry. Results Nimesulide effects a dose-dependent and time-dependent growth inhibition on QBC939 cells. High concentration of nimesulide (200 ?mol/L) not only inhibits the growth of QBC939 cells but also induces apoptosis cell nuclear condensation and apoptotic bodies were seen by transmission electron microscopy. Immunocytochemistry and RT-PCR shows upregulation of bax and down regulation of bcl-2 and survivin. Conclusion Nimesulide significantly inhibits the proliferation of QBC939 in vitro by induction of apoptosis in a dose- and time- dependent manner.

Objective: To understand the cardiac involvement of Dengue fever patients and to analyze the clinical characteristics of Dengue fever combining cardiac complications.
Methods: A total of 395 patients of Dengue fever treated in our hospital were analyzed. Blood levels of CK, CK-MB, CTnT, NT-proBNP were examined, ECG and echocardiography (UCG) were conducted in all patients. Upon Dengue recovery, the levels of CK and CK-MB were re-examined in patients with elevated indexes.
Results: Overall, there were 47.3% patients with increased CK, 61.3% with increased CK-MB, 42.5% with increased CTnT and 22.4% with increased NT-proBNP; 37.2% patients with abnormal ECG and 29.9% with abnormal UCG. There were 65.5% patients showing CK and CK-MB reduction more than 4 days upon Dengue recovery. Compared to those without the history of heart disease (HD), the patients with HD history had the higher abnormal rates of CTnT (17.7 vs 60.8) %, NT-proBNP (4 vs 32.3) % and ECG (25.1 vs 51.7) %, UCG (15.7 vs 41.7) %, allP<0.01. The abnormal rates in patients with heart failure (HF) were higher than those without HF as in CK-MB (75.8 vs 53.2) %, cTnT (85.5 vs 31.7) % and ECG (87.1 vs 31.7) %, UCG (63.8 vs 17.0) %, allP<0.01. The abnormal rates in elder patients were higher than non-elder patients as in CTnT (64.4 vs 7.7) %, NT-proBNP (32.2 vs 4.9) % and ECG (53.4 vs 21.8) %, UCG (38.9 vs 14.1) %, allP<0.01. The patients with elder age and HD history were more likely to have cardiac involvement upon Dengue infection.
Conclusion: Blood levels of CK, CK-MB, CTnT, NT-proBNP and ECG, UCG could be used to evaluate the patients with Dengue fever in order to prevent the cardiac complications at the early stage.

Objective To investigate the effects of nimesulide, a selective cyclooxygenase-2 (COX-2) inhibitor, on human cholangiocarcinoma QBC939 cell line in vitro. Methods The effects of nimesulide on QBC939 cells were observed with the following techniques: the influence of nimesulide on the proliferation of QBC939 cells was determined by MTT assay; the apoptosis of QBC939 cells was viewed and measured by transmission electron microscopy and flow cytometry, respectively; the expressions of proliferation cell nuclear antigen (PCNA) and COX-2 of cholangiocarcinoma cells were detected by immunocytochemistry. Results Nimesulide inhibited the expressions of PCNA and COX-2 and the proliferation of cholangiocarcinoma QBC939 cells, whose effects intensified as the dose increased and time elongated. Flow cytometry showed that the apoptotic rates of QBC939 cells increased significantly as the dose of nimesulide increased. The typical morphologic features of apoptosis were also observed by transmission electron microscopy. Conclusion Nimesulide significantly inhibits the proliferation of QBC939 cells in vitro by inducting cell apoptosis, which may be associated with the downregulation of COX-2 expression, and it also presents the features of dose and time dependents.

Objective:To observe the clinical effect of Tuina(Chinese therapeutic massage)combined with Bu Zhong Yi Qi Tang(Middle Jiao-supplementing and Qi-boosting Decoction)in the treatment of rectocele of Qi sinking due to spleen deficiency.Methods:A total of 108 patients with rectocele(grades Ⅰ and Ⅱ)were randomly divided into a Chinese medicine group and a Tuina+Chinese medicine group,with 54 cases in each group.Both groups were treated with Bu Zhong Yi Qi Tang,and the Tuina+Chinese medicine group was treated with additional Tuina manipulation.The score of clinical symptoms and grade of rectocele were observed and compared between the two groups.Results:After treatment,39 cases were cured,10 cases were effective,and 5 cases failed in the Tuina+Chinese medicine group,making a total effective rate of 90.7%.In the Chinese medicine group,26 cases were cured,16 cases were effective,and 12 cases failed,making a total effective rate of 77.8%.The efficacy difference between the two groups was statistically significant,checked by the rank-sum test(P＜0.05).After treatment,the grades of rectocele in the two groups were statistically different from those before treatment(P＜0.05),and the difference between the two groups was statistically significant(P＜0.05).After treatment,the score of each item and the total score of traditional Chinese medicine symptoms in the two groups were lower than those before treatment,and the differences within the group were statistically significant(P＜0.05).The score of each item and the total score of the Tuina+Chinese medicine group were lower than those of the Chinese medicine group,and the differences between the two groups were statistically significant(P＜0.05).Conclusion:Tuina plus Bu Zhong Yi Qi Tang has better clinical efficacy than Bu Zhong Yi Qi Tang alone in the treatment of rectocele of Qi sinking due to spleen deficiency.

Objective To explore the relationship between Y chromosomal microdeletions and chromosome karyotype,clinical phenotypes and sex hormone levels in patients with spermatogenic failure.Methods SRY gene,ZFY gene and 6 sequence-tagged-sites in AZFa,AZFb and AZFc were detected with multiplex polymerase chain reaction in 42 non-obstructive azoospermic and 39 severe oligozoospermic men.Results Ten cases (12.35%) of microdeletions were found in 81 non-obstructive azoospermic (6 cases) or severe oligozoospermic patients (4 cases).Among the 10 patients with Y chromosomal microdeletions, 1 had deletion of AZFa+AZFb+AZFc,2 AZFb+AZFc, 1 AZFb and 6 AZFc.Conclusion Microdeletions in the long arm of Y chromosome were associated with spermatogenic failure.Screening for Y chromosomal microdeletions in azoospermic or severe oligozoospermic patients should be of necessity and significance.

Objective To explore the effect and mechanism of Gli2 on EMT and invasion in the hepatocellular car-cinoma cell line SMMC-7721.Methods shRNA of Gli2 and Negative control (NC) shRNA were constructed and transfected into SMMC-7721 cells.shRNA-Gli2 group,shRNA-NC group and blank group were set up .Transwell chambers assay , adhesion experiments were used to detect the ability of invasion ,homogeneous and heterogeneous cells intercellular adhesion of each group .Meanwhile, the qRT-PCR, Western blot were used to examine Gli2, E-cadherin ,N-cadherin and vimentin mRNA and protein expression .Results In different hepatocellular carcinoma cell lines and be along with the increasing ability of the invasion in hepatocellular carcinoma cell lines , Gli2 expres-sion was higher .Compared with the shRNA-NC group and blank control group ,the interfered group extensive cells invasion ability was inhibited ( P <0.05 ) , homogeneous cells intercellular adhesion increased and heterogeneous cells intercellular was opposite ( P <0.05 ) , meanwhile , the expression of E-cadherin was declined significantly ( P<0.05 ) ,the expression of N-cadherin , vimentin raised significantly ( P<0.05 ) .Conclusions Silencing Gli2 gene can reverse the EMT of the hepatocellular carcinoma cell line SMMC-7721 and inhibit cell invasion ,its mecha-
nism may be related to the up-regulation of E-cadherin and the dow N-regulation of N-cadherin, vimentin.

The development of laboratory animal science plays a supporting role in the biomedical field, pharmacy and other fields.It is an important symbol to measure the level of science and technology in each countries in the world. The quality of employees directly affects the speed and quality of the development of laboratory animal industry.The purpose of this standard is to regulate the requirements of animal practitioners, to strengthen the training and qualification certification, and to promote the quality of employees.