Objective To evaluate the clinical significance of combined detection of phosphatidyhnositol 3 proteoglycans (GPC3) and alpha-fetoprotein heterogeneity (AFP-L3) and total bile acids (TBA) in diagnosis of primary hepatocellular carcinoma.Methods Collected 154 cases of primary hepatocellular carcinoma patients (hepatocellular carcinoma group) and 78 cases of cirrhosis patients (cirrhosis group) and 56 normal controls (control group) from May 2011 to December 2012.The level of GPC3,AFP-L3 was measured by enzyme-linked immunosorbent assay,the level of alpha-fetoprotein (AFP) was determined by mdioimmunoassay,and the level of TBA was measured by enzymatic cycle,and they were compared.Results The level of GPC3,AFP-L3 and TBA was (10.70 ± 3.10) μ g/L,(338.60 ± 379.20) μ g/L,(79.91 ± 70.64) μ mol/L in hepatocellular carcinoma group,which was higher than that in cirrhosis group [(2.70 ±0.71) μg/L,(6.45 ±2.79) μg/L,(33.10 ±21.90) μmol/L] and control group [(1.28 ± 0.60) μ g/L,(0.68 ± 0.56) μ g/L,(5.40 ± 2.20) μ mol/L],the level of GPC3,AFP-L3 and TBA in cirrhosis group was higher than that in control group,and there was significant difference (P＜ 0.01).The sensitivity of three tumor markers combined detection was 97.2％ (280/288),which was significantly higher than that of individual detection of GPC3 (72.1％,111/154),AFP-L3 (53.2％,82/154),TBA (94.8％,146/154),the difference was statistically significant (P ＜ 0.01).Conclusion Serum GPC3,AFP-L3 and TBA combined detection can improve the sensitivity of primary hepatocellular carcinoma and has important chnical significance in early diagnosis of primary hepatocellular carcinoma.

Objective To discuss the methods of the prevention of the superior laryngeal nerveinjury at endoscopic thyroidectomy through subclaviclar approach.Methods The clinical data of 100 patients with thyroid diseases received endoscopic thyroidectomy through subclavicular approach were analyzed retrospectively.Nodular goiter in 69 cases,adenoma in 18 cases,simple goiter in 13 cases.Line side lobe resection in 35 cases,the side lobe subtotal in 8 cases,the side lobe resection + subtotal contralateral lobe in 48 cases,bilateral subtotal gland in 9 cases.Results All 100 patients after surgery pronounced normal,no tone deep,choking water,etc,patients 7-30 d laryngoscope review found no abnormalities.Conclusion Thyroidectomy through subclavicular approach under endoscope is calleidic thyroidectomy,effectively avoid permanent superior laryngeal nerve injury.

Objective To investigate the expressions of Yes-associated protein-1 (YAP1) in gallbladder mucosal epithelium of normal persons,in patients with simple/calculous cholecystitis,and in patients with gallbladder carcinoma;and to study the mechanism of YAP1 in gallbladder carcinoma development.Methods Immunohistochemistry was used to detect the expression and distribution of YAP1 protein in 50 persons with normal gallbladder,101 patients with simple cholecystitis/calculous cholecystitis and 100 patients with gallbladder carcinoma.RT-PCR and western-blot were used to detect the mRNA and protein levels of YAP1 in normal and malignant gallbladder mucosal epithelium cells.siRNA was used to shut down the expression of YAP1 in SGC996 cells.MTT was used to test cell vitality.Flow cytometry was used to measure cell cycle.Results Immunohistochemistry revealed the expression rates of YAP1 in the gallbladder carcinoma group,the cholecystitis/gallstone group and the control group to be 87.0％ (87/100),56.4％ (57/101) and 5.0％ (1/20),respectively (P ＜ 0.01).The YAP1 protein levels were higher in gallbladder carcinoma tissues and cells when compared to normal tissues and cells.RT-PCR showed the mRNA levels of gallbladder carcinoma cells to be 12.5 ± 1.2 times of normal gallbladder mucosal epithelial cells (P ＜ 0.05).After using siRNA to shut down the YAP1 expression,EMT associated proteins were down-regulated,cell vitality was decreased,and cell cycle was arrested in the S-phase.Conclusions YAP1 is closely related to cell proliferation and metastasis of gallbladder carcinoma.It may promote tumor progression through epithelial-mesenchymal transition.transition;Tumor progress

Objective To establish the modified Ziehl-Neelsen acid -fast staining method and to investigate the value of modi-fied Ziehl-Neelsen acid-fast staining method of bronchoalveolar lavage fluid in the diagnosis of pulmonary tuberculosis(TB)with negative sputum specimen.Methods 50 cases of negative sputum pulmonary TB were performed the bronchoalveolar lavage by the fiberoptic bronchoscope before the treatment,at the same time,the bronchoalveolar lavage fluids were collected and detected by the two methods of the traditional and modified Ziehl-Neelsen staining.The diagnostic positive rates were compared between the two groups.Results The positive rates of the two kinds of acid-fast staining method were 38% and 82% respectively,the difference showing statistical significance(P <0.05 ).Conclusion The modified Ziehl-Neelsen acid-fast staining of bronchoalveolar lavage fluid can highly improve the positive diagnostic rate of pulmonary TB patients with negative sputum and deserves to be clinically promoted.

Objective:To investigate the expression of CXCL1 2 and CXCR4 in adenoid cystic carcinoma(ACC)and to explore its re-lationship with clinicopathologic characteristics and prognosis of the patients.Methods:The expression of CXCL1 2 and CXCR4 in af-fected tissue was detected immunohistochemically in 62 cases of ACC.Both of the two factors and clinicalpathology factors were pro-cessed in accordance with the Kaplan-Meier method and the COX regression model.Results:The positive rates of CXCL1 2 and CX-CR4 expression were 54.8%(34/62)and 77.42%(48/62)respectively.Patients with the 2 factor expression had a shorter survival time than those without them(P<0.05).Multivariate analysis revealed that CXCR4 expression,clinical stage,histological differentia-tion and metastasis/recurrence were independent risk factors for the prognosis of ACC patients.Conclusion:The expression of CXCR4 may be correlated with the malignancy of ACC.CXCR4 expression,clinical stage,metastasis/recurrence and histological differentia-tion can indicate the prognosis of ACC patients.

Objective To investigate the effect of doxapram on inhibition of medullary respiratory center excitability by sevoflurane in rats.Methods Neonatal Sprague-Dawley rats of both sexes,aged 1-4 days,were used in this study.Isolated medulla oblongata-spinal cord specimens were made according to the method described by Suzue and perfused with the artificial cerebrospinal fluid saturated with 95％O2-5％CO2.The specimens were randomly divided into 3 groups ( n =9 each):control group (group C),sevoflurane group (group S) and sevoflurane + doxapram group (group S + D).Respiratory rhythmical discharge activity of the hypoglossal nerve rootlets was recorded using suction electrode.After 10 min of equilibration,the specimens were perfused with the artificial cerebrospinal fluid,5％ sevoflurane and the mixture of 5％ sevoflurane and 5 μmol/L doxapram for 10 min in groups C,S,and S + D respectively.The respiratory cycle,inspiratory time and integral amplitude of inspiratory discharge were recorded.Results Compared with group C,the respiratory cycle was significantly prolonged,the inspiratory time was significantly shortened,and the integral amplitude of inspiratory discharge was significantly decreased in group S (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group S + D (P ＞ 0.05).Compared with group S,the respiratory cycle was significantly shortened,the inspiratory time was significantly prolonged,the integral amplitude of inspiratory discharge was significantly increased in group S + D ( P ＜ 0.05).Conclusion Doxapram antagonizes sevoflurane-induced inhibition of excitability of medullary respiratory center in rats.

Objective To investigate the effect of mixed-skin grafting with autologous microskin and allogenetic acellular dermal matrix microskin on wound healing in rats,and to make a further study on the related mechanism.Methods Wistar rats were served as a allogenetic acellular dermal matrix donor rats,and SD rats as acceptors with mould of full thickness skin defects on their back.The ninety SD rats were divided into 5 groups with 18 rats in each group.Group 1 was transplanted with autologous microskin,and group 2 with allogenetic acellular dermal matrix microskin.Groups 3,4 and 5 were grafted with mixed-skin ratio between autologous microskin and allogenetic acellular dermal matrix microskin 1 ∶ 1,1 ∶ 0.5 and 1 ∶ 0.25,repectively.The rate of wound healing was measured,wound samples collected,hematoxylin and eosin stain carried out,fibronectin (FN) and laminin (LN)detected,and intergroup comparison made,respectively,2,3 and 4 weeks after skin grafting.Results The wound healing rates and FN and LN expression of mixed-skin grafting groups were higher than those of the group with autologous microskin grafting.The group of 1 ∶ 0.25 obviously increased (P＜0.05 or P＜0.01).Conclusions The wound healing rate with mixed-skin grafting is higher than that with autologous microskin grafting.The best effect is achieved when the skin ratio between autologous microskin and allogenetic acellular dermal matrix microskin is 1 ∶ 0.25.It is possibly due to the increase of FN and LN on wound skin surface.

Objective To study the application of the combined detection of Golgi glycoprotein 73(GP-73),phosphatidylinositol proteoglycan 3(GPC3 )and percentage of AFP heteroplasmon(AFP-L3%)in the diagnosis of primary hepatocellular carcinoma (PHC).Methods The concentrations of GP-73,GPC3 and AFP-L3 were detected by enzyme-linked immunosorbent assay(ELISA) in 154 patients with PHC(PHC group),78 patients with cirrhosis(cirrhosis group)and 56 healthy subjects(control group).Then the detection results were statistically analyzed.Results The levels of GP-73,GPC3 and AFP-L3% in the PHC group were signifi-cantly higher than those in the liver cirrhosis group and the control group(P <0.05).The positive rates of GP-73,GPC3 and AFP-L3% in the PHC group were 66.2%,72.1% and 53.2% respectively.The positive rate in the combined detection of these three in-dices could reach 97.9%,which was higher than the sensitivity and accuracy in any single index detection and the combination de-tection.In the PHC group,the comparison between different levels of GP-73 and AFP-L3% with the AFP levels showed the statis-tically significant difference(P <0.05 ).Conclusion The combination detection of GP-73,GPC3 and AFP-L3% can improve the sensitivity and accuracy for diagnosing PHC and has reference significance in the differential diagnosis of early PHC.

Objective To explore the causation,diagnosis and management of iatrogenic bile duct injury(BDI) of laparoscopic cholecystectomy(LC).Methods A total of 1 656 patients undergoing laparoscopic cholecystectomy in our department during the last 7 years were included in this study for retrospective analysis.Results There were 274 patients with gallbladder polyps,168 patients with acute calculous cholecystitis and 1214 patients with chronic calculous cholecystitis.There were 15 BDIs associated with LC(0.91%).A total of 8 BDI patients were diagnosed during cholecystectomy.The remaining 7 BDI patients were diagnosed postoperatively.The intraoperative diagnosis of BDI was made on the discovery of bile leakage or double biliary stump during cholecystectomy.Clinical features,diagnostic abdominocentesis and imaging findings formed the basis of diagnosis of BDI postoperatively.One BDI patient was treated by repairing the injuried common bile duct with a T-tube drinage.Four BDI patients were treated by end-to-end anastomosis of injuried bile duct,and one of the four patients was reoperated with Roux-en-Y hepaticojejunostomy because of bile leakage.The remaining 10 BDI patients were treated by Roux-en-Y hepaticojejunostomy,and good results were achieved in all of these patients.Conclusions There is no relationship between the etiology of gallbladder disease and BDI during laparoscopic cholecystectomy.Good results can be achieved if BDI is diagnosed early and treated properly during or after operation.Roux-en-Y hepaticojejunostomy is the primary operation method for treating BDI.

AIM: To investigate the effects of mitochondrial ATP sensitive potassium (MitoK_ ATP) channel opening on translocation of protein kinase C epsilon (PKC?) and the relationship between the translocation of PKC? and the production of reactive oxygen species. METHODS: The expression of PKC? in cultured adult rat ventricular myocytes was investigated with immunofluorescence and Western blotting techniques. RESULTS: (1) Diazoxide, a selective MitoK_ ATP channel opener, caused a significant translocation to myofibrillar-like structures in cultured adult rat ventricular myocytes. (2) The N-2-mercaptopropionylglycine (MPG), a free radical scavenger, partly inhibited the translocation of PKC? caused by diazoxide. (3) Chelerythrine, a selective protein kinase C (PKC) inhibitor, completely blocked the translocation of PKC? caused by diazoxide. CONCLUSION: Opening of MitoK_ ATP channel might activate PKC? and make it translocation to myofibrillar-like structures. PKC? activation occurs downstream of MitoK_ ATP channel, and might be caused by production of reactive oxygen species after opening of MitoK_ ATP channel.