Objective To establish a HPLC method for quality control of Pei-bao capsule. Methods The content of lysine derivatized from 2,4-dinitoflruorobenzene was determined by HPLC, with chromatographic column being the agilent Zorbax SB-C18 column (250 mm×4.6 mm, 5 μm) , mobile phase being acetonirile-0.04mol/L, potassium dihydrogen phosphate (40 : 60), the flow rate being 1.0 ml/min, the column temperature being 30 ℃, and wavelength being 360 nm. Results It showed good linear coorelation when the concentration of Lysine hydrochloride being within 0.39～ 19.3μg/ml (γ =0.9999) , and the average sample recovery rate was 100.2%. Conclusion This method is simple, accurate, stable and reliable. It can be used for quality control of Pei-bao capsule.

Objective To investigate the influence of double-effect activation of cholinergic antiinflammatory pathway on the liver injury and inflammatory response in obstructive jaundice rats by applying cholinesterase inhibitor and cholinergic M receptor blocker to activate alpha 7 nicotinic acetylcholine receptor.Methods 22 adult male Wistar rats were randomly assigned into three groups:sham operation (SO) group (n=6),bile duct ligation (BDL) induced obstructive jaundice with (BDL treatment group) or without treatment (BDL control group) (n=8 each).The medicine treatment group was given anisodamine (25 mg/kg) and neostigmine (25 μg/kg) daily via intraperitoneal injection after surgery,the control group was given equal amount of normal saline.The body weights of rats in each group were measured every other day.After 12 days,the rats were killed,and the pathological changes of liver injury,liver function and the expression levels of proinflammatory cytokines in the serum and liver tissue were observed.Results The body weight of BDL rats was significantly lower than the SO group rats,and the growth rate of BDL treatment group rats was the same as the rats in BDL control group 3 days after the starting of treatment.The AST,ALT,bilirubin and gamma-GT levels of BDL control and treatment groups were significantly higher than the SO group (P＜0.05),but there was no significant difference between BDL control and treatment groups.The serum albumin level of BDL treatment group was obviously higher than that of BDL control group,but the pathological liver injury was significantly slighter.The gene expression levels of TNF-alpha,IL-1 beta and IL-6 in the liver tissue were significantly higher in BDL groups than SO group (P＜0.05),but BDL treatment group was significantly lower than BDL control group (P＜0.05).In addition the serum TNF-alpha and IL-1 beta concentrations of BDL treatment group and control group were significantly higher than the SO group (P＜0.05),but the BDL treatment group was obviously lower than that BDL control group (P＜0.05).Conclusion The combine application of cholinesterase inhibitor and cholinergic M receptor blocker to activate the cholinergic anti-inflammatory pathway can significantly inhibit the obstructive jaundice induced proinflammatory gene expression and liver injury.

Objective To establish a luciferase labeled McA-RH7777 hepatoma rat model,which could be used for gross observation to further observe the effect of selective ligation of the portal vein and bile duct on tumor growth and metastasis.Methods The luciferase gene was transfected into rat McA-RH7777 hepatoma cells with pCDH-puromycin-CMV as the carrier,which were subcutaneously inoculated into Buffalo rats.Tumor pieces were then heterotransplanted into the left lateral lobe of the allogenic rat liver to observe the tumor growth in vivo.After the successful hepatoma modeling,the rats were randomly divided into three groups,namely the implanted portal vein group with combined portal vein and bile duct ligation,the implanted portal vein group with single portal vein ligation and sham operation group.The rats were executed at the 1 st week and 2nd week after ligation,and the livers were dissected to record the tumor growth and metastasis inside and outside the liver,respectively.Results The tumor formation rates of Buffalo rats after subcutaneous and intrahepatic implantation were both 100％.The fluorescence signal implanted into the liver lobe could be observed in vivo after the intrahepatic implantation of luciferase transfected Luc-McA-RH7777 at 2nd week,the range and intensity of which increased over time.Only local tumor growth could be found at the 4th week,without obvious intrahepatic and lung metastasis.However,both an increased in situ tumor volume and the pulmonary metastasis could be observed in the implanted portal vein group with combined portal vein and bile duct ligation at 2nd week after the ligation.Immunohistochemistry showed AFP positive immunoreactions in the vast majority of intrahepatic tumor cells and Luc positive immunoreactions in part of tumor cells.Conclusion Luc-McA-RH7777 cells could be used to establish the heptoma rat model and the in vivo analysis within the Buffalo rat liver demonstrated that the combined ligation of the portal vein and bile duct can accelerate the development and metastasis of liver cancer.

Objective In order to improve cirrhotic liver management,each aspect of the liver's complex blood flow must be understood.This study investigates the protective effect of portal vein occlusion,with hepatic artery preservation,on cirrhotic liver after ischemia and reperfusion.Methods Carbon tetrachlorideand induced cirrhotic rats and normal rats were randomly assigned into 4 groups:normal sham operation (N-SO),cirrotic sham operation (C-SO),portal triad clamping (PTC),and portal vein clamping without hepatic artery inflow control (PVC).During the occlusion,the total 3-minute blood loss from the liver surface cut was weighed.At 1,6,and 24 hours post reperfusion,the serum alapine amino transferas (ALT),the adenosine triphosphate (ATP) of liver tissue,the malonolialdehgde (MDA) of liver tissue,and the morphological changes were evaluated.Result The amount of hemorrhage between the groups ranked as follows:PTC ＜ PVC ＜ N-SO ＜ C-SO (P＜0.05).At 1,6,and 24 hours post reperfusion.the ALT and MDA levels of the groups ranked as follows:PTC ＞ PVC ＞ C-SO ＞ N-SO (P＜0.05).Additionally,each group's ATP level ranked as follows:PTC ＜ PVC ＜ C-SO ＜ N-SO (P＜0.05).With histopathological examination,the hepatic injuries of the PTC and PVC group were more severe than those of the C-SO group,especially in the PTC group.Conclusion Therefore,the technique of portal vein clamping and hepatic artery inflow control can reduce the ischemic reperfusion injury of the cirrhotic rats' liver.

OBJECTIVETo observe the effect of congestion/reperfusion injury (CRI) and ischemia/reperfusion injury (IRI) on remnant liver in rats after hepatectomy.

METHODSMale SD rats were divided into IRI, CRI, and control groups. In the former two groups, the left lateral lobe of the rats were subjected to IRI or CRI for 30 min with the rest lobes (about 70% of the total liver weight) resected; the rats in the control group received hepatectomy preserving only the left lateral lobe. The mortality rate of the rats was recorded, and the surviving rats were sacrificed at 1, 3 and 7 days after operation for analyses of ICG plasma disappearance rate (ICG-PDR), ALT, AST, liver regeneration rate, and Ki-67 labeling index.

RESULTSThe mortality rate was significantly higher in CRI group (34.3%) than in IRI group (8%, P<0.05) and control group (4%, P<0.01). On day 1 following hepatectomy, CRI group showed significantly higher liver enzyme levels and poorer liver functions than the control group (P<0.05) without significant differences from those in IRI group (P<0.05); Ki-67 labeling index in CRI group was significantly lower than that in the control group (P<0.01) and IRI group (P<0.01). Compared with the control group, CRI group showed a significantly lowered maximum Ki-67 labeling index with also a delayed occurrence (P<0.01); CRI resulted in poorer liver regeneration rate on day 3 after hepatectomy compared to the control group (P<0.01) and IRI (P<0.05).

CONCLUSIONCompared with IRI, CRI can result in severer liver damage and lowered liver regenerative capacity in rats early after hepatectomy.

Animals ; Hepatectomy ; Liver Diseases ; Liver Regeneration ; Male ; Postoperative Period ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury

Objective: To investigate MAML2 gene-translocation in primary pulmonary mucoepidermoid carcinoma (PMEC) and pulmanary adenosquamous carcinoma, and the optimal diagnostic immunohistiochemical (IHC) panel in distinguishing PMEC from adenosqumous carcinoma. Methods: Twenty-four cases of PMEC and 44 adenosqumous carcinoma diagnosed in the Guangdong General Hospital were tested for MAML2 translocation by fluorescent in-situ hybridization (FISH) using tissue array. An IHC panel including TTF1, Napsin A, CK5/6, p63, p40 and Ki-67 was performed on the cohort. The clinical data for all cases were collected and all PMEC patients had follow-up information. Results: The patients′ age ranged form 6 to 73 years, with a median age of 32 years. The male to female ratio was 1.4∶1.0. MAML2 translocation was found in 16/24 (66.7%) cases of PMEC whereas all 44 cases adenosqumous carcinoma were negative for translocation. All the cases of the PMEC were negative for TTF1 and Napsin A but positive for CK5/6, p63 and p40 in the intermediate cells and epidermal-like cells. In most PMEC cases, the Ki-67 expression index was lower than 10%. In contrast, most cases of adenosqumous carcinomas expressed TTF1 and Napsin A in the adenomatous component and CK5/6, p63 and p40 in the squamous component, which expression pattern was different from that of PMEC. Based on IHC staining, 2 cases of highly invasive ALK-positive adenocarcinoma mimicing PMEC were also found in the study. Conclusions MAML2 gene translocation can be detected in about two-third of PMEC. Translocation of MAML2 gene and lower morphology grading are associated with good prognosis. The combined use of IHC antibodies panel is helpful to distinguish PMEC from the adenosqumous carcinoma and adenocarcinoma mimicing PMEC.

OBJECTIVETo study the clinical, pathologic, immunophenotype, molecular characteristics and prognosis of HIV-negative plasmablastic lymphoma (PBL).

METHODSTwelve cases of HIV-negative PBLs diagnosed between 2005 and 2014 in Guangdong General Hospital were identified according to WHO classification of tumors of haematopoietic and lymphoid tissues (2008). The clinicopathologic features and outcome were analyzed and the relevant literatures were reviewed.

RESULTSThe patients were predominantly male (11/12) with a median age of 55.5 years. The tumor cells showed the characteristic combination of immunoblastic/plasmablastic morphology, plasma cell phenotype and high proliferation, no expression of mature B cell markers. 7/10 of the cases were EBER positive. Two cases were positive for C-myc translocation. Four of twelve patients were died.

CONCLUSIONSPBL is a rare, aggressive B-cell lymphoma. HIV-negative PBL has lower rate of oral involvement and EBER expression than HIV-positive patients, the differential diagnosis is very challenging, and the prognosis is worse.

Diagnosis, Differential ; Female ; HIV Seronegativity ; Humans ; Immunophenotyping ; Lymphoma, B-Cell ; diagnosis ; Male ; Middle Aged ; Plasma Cells ; classification ; Plasmablastic Lymphoma ; diagnosis ; pathology ; Prognosis ; Translocation, Genetic

Objective To investigate the difference of hepatic microvessel density, neovasculariza-tion of regenerating liver tissue after ablation of two ways of irreversible electroporation and radiofrequency ablation in rats. Methods 90 male Sprague-Dawley rats were randomly divided into 3 groups, including the control group ( n =30), the irreversible electroporation group ( n =30 ) and the radiofrequency ablation group (n=30). 3,7 and 10 days were executed after the operation and draw material, expression of vascu-lar endothelial growth factor(VEGF) and CD34 in tissue was studied by immunohistochemistry, and the mi-crovascular density of tissue and VEGF positive cells were measured. Results The microvascular density of 3, 7 and 10 days in the control group was 50. 3 ± 12. 5, 54. 6 ± 11. 9 and 58. 2 ± 14. 7, the microvascular density of the radiofrequency ablation group was 18. 4 ± 4. 7, 17. 3 ± 5. 1 and 18. 1 ± 5. 9, respectively. The microvascular density of the irreversible electroporation group was 42. 8 ± 10. 4, 45. 6 ± 10. 2 and 49. 2 ± 13. 8, respectively. The positive cells of VEGF in control group was 50, 56 and 57 at 3, 7 and 10 days, and 32, 30 and 33 at 3, 7 and 10 days in radiofrequency ablation group, 44, 43 and 45 at 3, 7 and 10 days in irreversible electroporation group; expression of VEGF and CD34 in 3, 7, 10 d and the microvascular density of ablation area in radiofrequency ablation group was significantly lower than those in control group after irreversible electroporation and radiofrequency ablation. No significant differences were found between irreversible electroporation group and control group. Conclusion The irreversible electroporation can effectively protect the microvessels in the ablation area, ensure the tissue’s blood supply after the ablation, and provide a guarantee for the repair and regeneration of the tissue.

Objective:To establish a new bile duct injury and repair model in mice by generating bile duct distal stricture and proximal dilatation.Methods:The mice were randomly divided into sham operation group, bile duct stricture (BDS) group and bile duct ligation (BDL) group. The dilated bile duct of BDS mice was injured and then repaired 14 days after the modeling operation. Biochemical markers were detected and histopathological changes were analyzed.Results:14 days after the establishment of the model, the body mass in BDL group was significantly lower than that of the sham group ( P<0.05), while the body mass in BDS group was similar to sham group. Compared with the sham group, the bile duct and gallbladder of the BDS group and BDL group were both prominently dilated, but the sum of the diameters of bile duct and gallbladder in BDS group was significantly smaller than that in the BDL group ( P<0.05). Indocyanine green fluorescence imaging confirmed that biliary tract of BDS group could still drain bile. Serum ALT, AST and TBil levels in the BDS group were slightly higher than those in the sham group (all P<0.05), but significantly lower than those in the BDL group ( P<0.05). Bile ducts of BDS mice were injured by notching and repaired with bile duct path. 30 days after the repairing, HE staining showed that the bile duct epithelium around the patch was arranged in orderliness. Immunohistochemistry confirmed the positive staining of green fluorescent protein (EGFP) and CK19 in those groups. Conclusion:This model of bile duct injury and repair in mice can provide a new model for the study of the mechanism of bile duct injury and repair and the evaluation of tissue engineering bile duct.

Objective:To determine the risk factors of drainage time longer than 1 day in patients with selective abdominal drainage after laparoscopic cholecystectomy.Methods:The clinical data related to patients with selective abdominal drainage undergoing laparoscopic cholecystectomy from November 2009 to November 2019 at Chinese PLA General Hospital were retrospectively analyzed. Of 233 patients enrolled into this study, there were 147 males and 86 females, with a median aged 59.0 (47.5, 65.5) years old. The patients were divided into drainage time 1 day group of 65 patients and longer than 1 day group of 168 patients according to postoperative drainage time. The baseline data and perioperative data were collected, the risk factors correlated with drainage time longer than 1 day were analyzed.Results:The drainage time was 1 in the 1 day group and 2~8 in another group. Among the 233 patients, there was one with biliary leakage and 14 patients had abdominal bleeding, all of them healed after 2~3 days. All of the 233 patients were recovered when discharged. Independent risk factors related to drainage time longer than 1 day include BMI&ge;28 kg/m 2 ( OR=3.443, 95% CI: 1.411-8.405, P=0.007), operation time &ge;65 min ( OR=2.570, 95% CI: 1.310-5.045, P=0.006), thickness of gallbladder wall &ge;0.5 cm ( OR=12.720, 95% CI: 1.350-5.478, P=0.005), postoperative stomachache ( OR=13.537, 95% CI: 1.685-108.748, P=0.014) and postoperative fever ( OR=8.156, 95% CI: 1.035-64.249, P=0.046). Conclusion:For patients undergoing selective abdominal drainage after laparoscopic cholecystectomy with BMI &ge;28 kg/m 2, operation time &ge;65 min, gallbladder wall thickness &ge;0.5 cm, postoperative abdominal pain and fever, clinicians should appropriately prolong the drainage time to ensure medical safety.