Objective To explore the management of unsuspected gallbladder carcinoma (UGC) discovered during laparoscopic cholecystectomy (LC). Methods From January 2000 to May 2007, 15 560 cases of LC were performed in our hospital, UGC was discovered in 28 of the cases. Clinical data of the 28 patients were analyzed retrospectively. Results Among the 28 cases of UGC, 12 were discovered intraoperatively, and 16 were diagnosed postoperatively.Radical cholecystectomy was performed on the 12 cases who were detected during the operation,and 5 of the 16 patients who were diagnosed after the operation.LC was carried out in the remaining 11 cases. Postoperative pathological examination showed TNM stageⅠin 3 cases, stageⅡin 23, and stage Ⅲ in 2. The patients were followed up for 7-66 months with a mean of 22 months. During the period, 19 patients died. The 1-, 3-, and 5-year survival rate of the 17 patients who received radical cholecystectomy were 76.5% (13/17), 23.5% (4/17), and 11.8% (2/17), respectively. Whereas, in the 11 patients who underwent LC, the 1-year survival rate was 45.4%; moreover, none of the 11 survived for more than 2 years. Conclusions For high-risk population, attention should be paid to UGC during operation. Most of the UGCs discovered during LC are early-stage carcinomas, which should be treated with standard radical cholecystectomy, but not LC.

Objective To discuss how to improve judgement and technique for avoiding bile duct injury during laparoscopic cholecystectomy(LC).Methods The clinical data of 39 860 patients treated by LC between October 1992 and October 2005 were analyzed retrospectively.Bile duct injury occured in 46 patients.Results Bile duct repair and T-tube drainage was performed in 26 patients and bile duct end-to-end anastomosis and T-tube drainage was performed in 4 patients,with the T-tube indwelling time for 3~12 months.The Roux-en-Y cholangiojejunostomy was conducted in 11 patients,the ligation of accessory hepatic duct was conducted in 5 pauients,the re-operation for bile duct stenosis in 4 patients,and the re-operation for stenosis after cholangiojejunostomy,in 2 patients.Conclusions Intensive anatomic knowledge and skillful surgical performance can avoid or minimize the incidence of bile duct injury.Early detection and active repair of the bile duct injury for preventing acute inflammation is the most important measure to avoid repeated operations.

Objective To investigate the effects of Huaier Granule combined with TAC chemotherapy on immunologic function and prognosis in triple negative breast cancer patients after operation.Methods Ninty-two cases of Ⅰ-ⅢA stage triple negative breast cancer patients entered the study.They were randomly divided into control group and research group.Only TAC chemotherapy was affected in control group of 42 cases, while Huaier Granule combined with TAC chemotherapy was applied in research group of 50 cases.And we gave Huaier Granule to patients in the first day of chemotherapy.The changes of T cell subset, NK cell were detected in the patients respectively in the first day before therapy and after therapy.The five-year disease-free survival rate and overall survival rate were also observed in two groups.Results Compared with control group, the changes of T cell subset, NK cell had no significant differences in research group in the first day before therapy(P ＞0.05).In the first day after therapy, the level of CD4 + (38.16 ± 5.71) ％ in control group was significantly lower than that (44.67 ± 6.47) ％ in research group(P ＜ 0.05).However, the percentage of CD8 + T cells (28.68 ± 4.06) ％ in control group was markedly higher than that (22.85 ± 3.22) ％ in research group (P ＜ 0.05).The level of NK cell (15.75 ± 3.47) ％ in control group was obviously lower than that (19.46 ± 4.22) ％ in research group(P ＜ 0.05).The five-year disease-free survival rate is 28.6％ in control group, however the five-year disease-free survival rate is 42.0％ in research group, there were significant differences between two groups(P ＜ 0.05).Furthermore, the five-year overall survival rate in research group was obviously higher than that in control group(P ＜ 0.01).Conclusions Huaier Granule combined with TAC chemotherapy not only enhances immune function but also improves prognosis and quality of life in triple negative breast cancer patients after operation.It provides a positive exploration for anti-cancer research by integration of traditional and western medicine.

Aim To explore the effects of tetrameth-ylpyrazine-2′-O-sodium ferulate (TSF)on the swelling of
astrocytes and the expression of AQP4 after oxygen gl-ucose deprivatio /reoxygenation(OGD /Reox).Methods
Astrocytes were divided into 4 groups:control group, OGD /Reox group,Ozagrel group and TSF group.The effects of TSF on astrocytes were investigated 6,1 2,24 and 48 h after OGD /Reox.The cell injury was assessed by measuring LDH activity and MTT.The expression levels of AQP4 protein of astrocytes were detected u-sing Western blot.Results OGD /Reox induced obvi-ous cell swelling and significant reduction of LDH in astrocytes whereas TSF remarkably attenuated OGD-in-duced astrocyte swelling and LDH reduction (P <0.01 ).The expression levels of AQP4 protein in-creased gradually to the peak at 48 h in OGD /Reox
group(P <0.05 ).AQP4 protein levels in TSF group were much lower compared with OGD /Reox group at each time point (P <0.05).And there was no signifi-cant difference between TSF group and OGD /Reox group(P >0.05 ).Conclusion TSF can attenuate OGD-induced swelling of astrocytes through decreasing the AQP4 expression.

Objective To discuss about the relationship between the occurrence of the uterine cervix cancer and the risk factors.MethodsAmong the patients we collected,92 patients were diagnosed to have the cervical cancer,63 patients were diagnosed as CINⅢ (the cervical intraepithelial neoplasia,CIN),and 252 patients belonged to the control group.All patients had not been cured before.We used the questionnaire to investigate them about the dietary habit and other information.Through using different statistical Methods ,we explored the relationship between the cervical cancer and the potential factors.ResultsWe found out that many factors contributed to the happening of the cervical cancer,such as gynecopathy(χ2=19.31,P＜0.01),the first time of the menses(F＝11.62,P＜0.01),the first time of the sex life(F＝25.76,P＜0.01),the first time of the parturition (F＝28.02,P＜0.01),the times of the pregnancy and the parturition(F1＝13.98,P1＜0.01;F2＝4.78,P2＜0.01),the culture degree(χ2＝10.70,P＜0.05),the infection of HPV(χ2＝179.95,P＜0.01),the level of the folic acid (F＝3.39,P＜0.05) and so on.The result of the logistic regression analysis showed that keeping the habit of drinking tea(OR＝0.321,β＝-1.136,P＜0.05),paying attention to the sanitation of the sex life(OR＝0.387,β＝-0.950,P＜0.05),putting off the first time of the sex life(OR＝0.551,β＝-0.596,P＜0.05),and the higher level of the folic acid (OR＝0.502,β＝-0.688,P＜0.05) were the protective factors tothe occurrence of the uterine cervix cancer.Meanwhile,the infection of the HPV(OR＝27.215,β＝3.304,P＜0.01),many times of the parturition(OR＝1.846,β＝0.613,P＜0.05) and the passive smoking (OR＝1.673,β＝0.515,P＜0.05) were the risk factors.ConclusionsWith the higher level of the folic acid,the less possibility you will get the cervical cancer.There exists many measures to prevent the happening of the cervical cancer,like keeping the healthy sex life,keeping far away from the passive smoking,not to give birth too early or too many and preventing being infected the HPV.What's more,having the good habit of drinking tea and paying attention to the supplement of the folic acid and Vitamin B12 may be the effective method to prevent the occurrence of the cervical cancer.

Nowadays, proteomics focuses on quantitative analysis rather than qualitative. In the field of quantitative proteomics, Isobaric tags for relative and absolute quantitation (iTRAQ) is one of the most widely used techniques. The advantage of iTRAQ is high throughput, high stability and free of the restriction of sample property. iTRAQ is suitable for almost all kinds of samples, and up to 8 samples can be analyzed simultaneously by commercially available kit. Along with the development of techniques, more and more mass spectrometry (MS) platforms are used in iTRAQ experiments and the accuracy of iTRAQ has been improved. iTRAQ has been applied to studies of microorganism, animal, plant, medical and protein post-translational modification. Here we review the recent progress in the development of iTRAQ and its applications in quantitative proteomics.
Animals ; Humans ; Mass Spectrometry ; Proteomics ; methods

The phospholipase A2 ( PLA2 ) is one of the main constituents of the bee venom. Its hypotensive effect and mechanism are studied in this report.In the anaesthetized rats and cats, the PLA2 of the bee venom given intravenously caused a quick and profound fall in the arterial blood pressure.The results of this study indicate that its hypotensive effect is mainly concerned with releasing the endogenous histamine. If the antagonists of H1 and H2 receptor are administered together, its hypotensive action will be countered.

Objective To construct human Cystatin C expression vector and make primary protein purification. Methods A 380 bp cDNA fragments encoding Cystatin C were isolated by RT PCR from human placenta tissue and then inserted into pPIC9 vector. Recombinant pPIC9 Cystatin C was transformed into Pichia pastoris strain GS115. Secreted soluble Cystatin C was produced by induction of the AOX1 promoter with methanol. Recombinant Cystatin C was purified by Hitrap SP cation exchange chromatography. Results The expression level of Cystatin C reached 16 mg/L. The stability of Cystatin C is 3 days in expression system. SDS PAGE revealed that Cystatin C was expressed up to 60% of the total soluble protein. After purified by Hitrap SP cation exchange, the recovery rate was 40.5%. Conclusion Cystatin C has a high secretion in Pichia pastoris yeast and could be recognized by polyclonal antibody of native human Cystatin C. After purification,this protein can be used as a standard and antigen to develop immunoassay for human Cystatin C in serum.

Objective To explore the method of obtaining and expanding epithelial stem cells(ESCs) from plucked hair follicle in vitro and clarify that fibroblasts are necessary for ESCs growth.Methods A amount of ESCs from plucked hair were obtained,the epithelial outgrowth of human plucked hair follicle was studied and compared between different culture conditions including no coating well,rat tail-collagen coating well,collagen Ⅳ coating well and 3T3 fibroblasts feeder.The characteristics of these cells were detected by RT-PCR and immunofluorescent staining for CK15.Results After compared between different culture conditions including no coating well,rat tail-collagen coating well,collagen Ⅳ coating well and 3T3 fibroblasts feeder,the results indicated that the plucked hair fragment B3(bulge area of hair follicle) placing on 3T3 fibroblasts feeder was the optimum condition.RT-PCR and immunofluorescent method confirmed that these cells expressed CK15 which was the special marker of ESCs.Conclusion ESCs can be obtained from bulge area of hair follicle which grows on the 3T3 fibroblasts feeder and this method is the optimum condition for ESCs growth after compared between different conditions.

Objective To develop a dual real-time fluorescent RT-PCR method for rapid detection of enterovirus(EV)and en terovirus type 71(EV71).Methods Specific primers and probes were designed and the dual real-time fluorescent RT-PCR reaction system was established.The quantitative standard curve was drawn;its sensitivity and precision were evaluated.Feces and throat swab specimens of 109 clinical patients with hand foot and mouth disease were collected and tested by using this method.Then the obtained results were compared with those detected by commercial EV71 PCR kit.Results The relative coefficient(2)of EV and EV71 standard curve established by the dual real-time fluorescent RT-PCR method were both 0.998.Its sensitivity reached 0.5 TCID50/mL for detecting EV and 0.05 TCID50/mL for detecting EV71.The within-run precision for detecting EV and EV71 was ＜3％ and total precision≤4％.The results showed good specificity for the detection of enterovirus and non-enterovirus.In 109 detected clinical samples,84 cases of EV positive samples were detected,in which 56 cases were EV71 positive with the total positive rate of 51.4 ％,which was consistent with the result of simple fluorescent RT-PCR commercialization kit(P=1.000).Conclusion The established dual real-time fluorescent RT-PCR method has high sensitivity and good stability,which has an important significance for early high throughput rapid diagnosis of hand foot and mouth disease.