Objective To study the prevention and treatment of bile duct injury during laparoscopic cholecystectomy(LC). Methods 22 cases of bile duct injury during LC were reviewed retrospectively. The features, diagnosis, treatment and efficacy of injury were summarized. Results All patients were successfully treated by Roux-en-Y cholangio-jejunostomy. Besides, 8 patients underwent plastic operation of bile duct of hepatic portal and 3 patients middle lobectomy of liver. 22 cases were followed up at the 1st and 3rd year after surgery, and no bile duct stricture, recurrence of jaundice and cholangitis occurred. Conclusions It's a key to prevent bile duct injury during LC. The management of bile duct injury should be chosen according to injured time, sites and types.

Objective To assess the levels of glucose-6-phosphate isomerase(GPI) mRNA in peripheral blood monocytes and serum GPI levels in patients with rheumatoid arthritis, and analyze the association of serum GPI with MCV antibody, CCP antibody and RF of RA. Methods Fluorogenic quantitative real-time polymerase chain reaction (FQ-RT-PCR) was used to examine mRNA expression on peripheral blood monocytes in 60 RA patients (28 case in active stage,32 cases in stable stage) ,30 patients with other rheumatic diseases, and 30 healthy controls. ELISA was used to detect the levels of serum GPI, anti-MCV antibodies, anti-CCP antibodies and RF in each group. Results The levels of GPI mRNA in RA group [△Ct=4.21 (3.04-7.23)] were significantly higher than those in patients with other rheumatic diseases [△Ct=8.42 (5.16-9.98),P<0.01] and healthy controls [△Ct=8.66 (4.90-10.01), P<0.01]. There were statistically significant differences of GPI mRNA levels between active RA [△Ct=3.78 (1.28-6.09)] and inactive RA[△Ct =5.88(3.23-8.94),H=11.760,P<0.01)]. The RA group serum GPI levels [3.02 (2.02-8.39) mg/L] were higher than those of other rheumatic diseases [0.20 (0.11-0.32) mg/L] and healthy controls [0.18(0.08-0.30) mg/L]. There were significant differences of serum GPI levels between active RA group [4.84(2.81-10.38) mg/L] and inactive RA group[2.12 (1.26-4.34) mg/L] (H=9.830, P<0.01). The sensitivities of GPI, anti-MCV and anti-CCP were 68% (41/60) ,57% (34/60),58% (35/60), respectively and specificities were 95% (57/60), 92% (55/60) and 93% (56/60), respectively. Conclusions The high expression of GPI mRNA in RA patients shows that it may play a pathological role in the development of RA, and it may be correlated with the activity of RA. It may be a valuable diagnostic parameter for RA, because of its high sensitivity and specificity.

Objective To explore the effect of radiofrequency ablation (RFA) combined with transcatherter arterial chemo-embolization (TACE) and percaulaneous ethanol injection (PEI) for unresectable hepatic malignancies. Methods The clinical data of 41 cases of unresectable liver cancer(URLC)treated by RFA,TACE and PEI were analysed retrospectively. Results There were 30 cases of primary hepatic cancer(PHT) and 11 cases of secondary hepatic cancer in this series.Ultrasound,CT and MRI showed the tumors shrinking or stable in 26 of the 41 patients postoperatively.AFP decreased to normal in 12 cases of 16 AFP positive PHT patients after operation. No severe complication was seen in the series. Conclusions RFA combined with TACE and PEI is a safe, well-tolerated and effective method for unresectable hepatic carcinoma, and may improve the treatment efficacy of URLC.

Objective Reverse transcription-polymerase chain reaction (RT-PCR) is widely used in gene expression analysis.Selection of proper housekeeping gene is very important.Rat model of myocardial infarction is a major animal model of myocardial infarction.Few reports are found about the selection of rat housekeeping gene after myocardial infarction.This study was aimed to explore the stability of the housekeeping genes expression in the rat model of myocardial infarction.Methods Myocardial infarction models were made using the anterior descending coronary artery ligation method.Combining with practical work,analysis and selection were made of four widely used standard housekeeping genes:glyceraldehydes-3-phosphate dehydrogenase (GAPDH),ribosomal protein L13A (RPL13A),beta-actin (ACTB) and acidic ribosomal phosphoprotein P0 (ARBP).The expressions in the rat heart were compared using real-time fluorescent RT-PCR and analysis was carried out to figure out which was the most suitable housekeeping gene for study of gene expression in rats after myocardial infarction using GeNorm program.Results The M values of RPL13A,GAPDH,ARBP and ACTB gene were 0.812,0.721,0.812 and 1.2 respectively.Conclusion GAPDH and ARBP are the most stable genes for the rat myocardial infarction model.

Objective To investigatc the role of phosphatidylinositol 3-kinase (PI3K)/protein-serine-threonine kinases (Akt) signal pathway in ginsenoside Rb1 pretreatment-induced attenuation of myocardial ischemiareperfusion (I/R) injury in diabetic rats.Methods Male SD rats weighing 250-300 g were used in this study.Diabetes mellitus was induced by intraperitoneal streptozotocin and confirmed by fasting blood glucose ≥ 16.7mmol/L.Eight weeks after diabetes mellitus was induced,48 rats were randomly divided into 4 groups ( n =12each):group myocardial I/R (group I/R); group ginsenoside Rb1 (group R); group ginsenoside Rb1 + wortmannin (PI3K inhibitor) (group RW) and group wortmannin (group W).Myocardial I/R was induced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion.Ginsenoside Rb1 40 mg/kg was injected iv at 10 min before ischemia in groups R and RW,while in groups RW and W wortmannin 15 μg/kg was injected iv at 20 min before ischemia.Arterial blood samples were collected at the end of 120 min reperfusion for determination of creatine kinase (CK) and lactate dehydrogenase (LDH) activities.The rats were then sacrificed.The infarct size was measured by tetrazolium method.Myocardial apoptosis was detected by TUNEL and apoptotic index (the number of apoptotic myocardial cells/the total number of myocardial cells) was calculated.The expression of Akt and phosphorylated Akt (p-Akt) was determined by Western blotting.Results Ginsenoside Rb1 pretreatment significantly reduced the infarct size,myocardial cell apoptotic index and serum CK and LDH activities and up-regulated p-Akt expression in group R as compared with group I/R.The protective effects of ginsenoside Rbl against myocardial I/R injury were significantly attenuated by wortmannin pretreatment in group RW compared with group R.Conclusion PI3K/Akt signal pathway is involved in the protective effects of ginsenoside Rb1 against myocardial I/R injury in diabetic rats.

Objective To evaluate the role of phosphatase and tensin homologue deleted on chromosome 10 (PTEN) in diabetes mellitus-induced reduction of hypoxic postconditioning (HPO)-induced protection of cardiomyocytes and the relationship with glycogen synthase kinase-3β (GSK-3β)-mediated mitochondrial apoptotic pathway.Methods H9c2 cells incubated in high-glucose (30 mmol/L) medium for 24 h were divided into 6 groups (n =5 each) using a random number table:normoxia group (group N),hypoxia-reoxygenation (H/R) group,group HPO,PTEN gene silencing normoxia group (group P-N),PTEN gene silencing H/R group (group P-H/R),and PTEN gene silencing HPO group (group P-HPO).H9c2 cells were exposed to 95％ N2-5％ CO2 for 4 h followed by 2 h reoxygenation with 90％ O2-10％ CO2.HPO was induced by 3 cycles of 5 min reoxygenation followed by 5 min hypoxia before reoxygenation.At the end of reoxygenation,the level of lactate dehydrogenase (LDH) in the supernatant was detected by enzyme-linked immunosorbent assay,the changes in mitochondrial membrane potential (MMP were assessed by JC-1 fluorescence assay,the cell apoptosis was detected by AnnexinV-FITC/PI flow cytometry,and the expression of PTEN and phosphorylated GSK-3β (p-GSK-3β) was determined by Western blot.The JC-1 monomer/polymer ratio and apoptosis rate were calculated.Results Compared with group N,the amount of LDH released,JC-1 monomer/polymer ratio and apoptosis rate were significantly increased,and the expression of PTEN was up-regulated in H/R and HPO groups (P＜0.05).There was no significant difference in the parameters mentioned above between group H/R and group HPO (P＞0.05).Compared with group HPO,the amount of LDH released,JC-1 monomer/polymer ratio and apoptosis rate were significantly decreased,PTEN expression was down-regulated,and the expression of p-GSK-3β was up-regulated in group P-HPO (P＜0.05).Compared with group N,the expression of PTEN was significantly down-regulated,and no significant changes were found in the other parameters mentioned above in group P-N (P＞0.05).Compared with group H/R,the expression of PTEN was significantly down-regulated,and no significant changes were found in the other parameters mentioned above in group P-H/R (P＞0.05).Conclusion PTEN is involved in diabetes mellitus-induced reduction of HPO-induced protection of cardiomyocytes,and the mechanism is associated with PTEN-induced activation of GSK-3β-modulated mitochondrial apoptotic pathway.

Objective To investigate the effects of penehyclidine hydrochloride (PHC) on acute lung injury induced by blunt chest trauma-hemorrhagic shock and resuscitation in rats.Methods Forty male SpragueDawley rats,aged 8 weeks,weighing 250-300 g,were randomly assigned into 4 equal groups (n =10 each) using a random number table:sham operation group (S group),blunt chest trauma combined with hemorrhagic shock and resuscitation group (group THSR),PHC for prevention group (group P1)and PHC for treatment group (group P2).ALI was induced by dropping a 300 g weight onto a precordial protective shield to direct the impact force away from the heart and toward the lungs in anesthetized rats in THSR,P1 and P2 groups.Blood was withdrawn via the femoral artery 5 min later until MAP was decreased to 35-45 mmHg within 15 min and maintained at this level for 60 min,followed by resuscitation.In P1 group,PHC 2 mg/kg was injected intravenously at 30 min before blunt chest trauma.In P2 group,PHC 2 mg/kg was injected intravenously at 60 min after hemorrhagic shock.At 6 h after the model was established,arterial blood samples were obtained for blood gas analysis and for measurement of concentrations of interleukin-6 (IL-6) and interleukin-1β (IL-1β) in serum by ELISA.Oxygenation index (OI) was calculated.The animals were sacrificed and bronchoalveolar lavage fluid (BALF) was collected for determination of white blood cell count and protein concentrations.Lungs were removed for examination of pathological changes and ultrastructure and for determination of Toll-like receptor (TLR4) and phosphor-p38 mitogen activated protein kinase (p-p38MAPK) expression (by Western blot).Results Compared with group S,PaO2 and OI were significantly decreased,PaCO2,protein concentrations in BALF,white blood cell count,and IL-6 and IL-1β concentrations in serum were increased,and TLR4 and p-p38MAPK expression was up-regulated in THSR,P1 and P2 groups.Compared with group THSR,PaO2 and OI were significantly increased,PaCO2,protein concentrations in BALF,white blood cell count,and IL-6 and IL-lβ concentrations in serum were decreased,TLR4 and p-p38MAPK expression was down-regulated in P1 and P2 groups.No significant differences were found in the parameters mentioned above between P1 and P2 groups.Conclusion PHC can mitigate acute lung injury induced by blunt chest trauma-hemorrhagic shock and resuscitation in rats,and inhibited activation of TLR4/ p38MAPK signaling pathway and attenuated inflammatory responses are involved in the mechanism.

Objective To investigate the effects of penehyclidine hydrochloride on Fas/FasL expression during acute lung injury induced by blunt chest trauma-hemorrhagic shock and resuscitation (HSR) in rats.Methods Thirty male SPF Sprague-Dawley rats, aged 8 weeks, weighing 245-275 g, were randomly assigned into 3 equal groups using a random number table: sham operation group (group Sham) , blunt chest trauma-HSR group (group THSR) and penehyclidine hydrochloric group (group PHCD).The model of acute lung injury induced by blunt chest trauma-HSR was induced by dropping a 300 g weight onto a precordium in anesthetized rats.Blood was withdrawn via the femoral artery 5 min later until mean arterial pressure was decreased to 35-45 mmHg within 15 min, and maintained at this level for 60 min, followed by resuscitation.In PHCD group, PHCD 2 mg/kg was injected intravenously at 60 min after hemorrhagic shock.At 6 h after the model was established, the rats were sacrificed, the lungs were then removed for microscopic examination of pathologic changes and for determination of Fas, FasL and caspase-8 expression, and interleukin-6 (IL-6) and IL-1β contents in lung tissues.Apoptotic index was calculated.Results Compared with group Sham, the expression of Fas, FasL and caspase-8 was significantly up-regulated, and AI and contents of IL-6 and IL-1β were increased in THSR and PHCD groups (P＜O.05).Compared with group THSR, the expression of Fas, FasL and caspase-8 was significantly down-regulated,and AI and contents of IL-6 and IL-1β were decreased in group PHCD (P＜0.05).The pathologic changes of lungs were significantly reduced in group PHCD compared with group THSR.Conclusion The mechanism by which penehyclidine hydrochloride inhibits lung cell apoptosis induced by blunt chest trauma-HSR is associated with inhibition of Fas/FasL expression in rats.

Objective To analyze the differentially expressed proteins in bone marrow supernatants of multiple myeloma patients by using 2-DE and MALDI-TOF-MS,and search for the special protein markers for studying the mechanism of the development and diagnosis or differential diagnosis of multiple myeloma.Methods The bone marrow supernatant samples of fourteen multiple myeloma patients,five other hematologic malignancies and five normal controls were collected.After removing albumin and IgG,the proteins in the supernatants were separated by 2-DE.Three groups images were analyzed and compared by Imagemnster 2D platinum 5.0 analysis software.Differentially expressed proteins were selected if the protein spots intensity showed more than 3 fold increase or decrease among different groups.The identities of the differentially expressed proteins with good repeatibility were determined by PMF based on by MALDI-TOF-MS or MALDI-TOF-MS/MS and NCBInr database search.Results 2-DE maps of bone marrow supernatants of the three groups could be analyzed and compared by image analysis of software.Forty-seven and fifty-eight differentially expressed protein spots were detected in multiple myeloma samples compared with normal controls and other hematologic malignancies samples,respectively.Forty-one reproducible spots were analyzed and identified by mass spectrum.Compared with other hematologic malignancies and normal controls,five up-expressed proteins and three down-expressed proteins were identified in multiple myeloma samples.They includes immunoglobulin J chain κ and λ light chain,provirus ancestral Gag polyprotein,mature oxy-cope catalytic antibody with hapten for up-expressed proteins,and hemoglobin,haptoglobin Hp2,zinc-alPha-2-glycoprotein for down-expressed proteins.These differentially expressed proteins reflect the features of muhiple myeloma,and relate to the development,progression and therapy of multiple myeloma.Conclusions Eight differentially expressed proteins in bone marrow supernatants of multiple myeloma are identified by using 2-DE and MALDI-TOF-MS.These differentially expressed proteins could be useful in studying the mechanism of the development and progression of multiple myeloma,and in developing diagnosis and differential diagnosis of multiple myeloma.

Objective To evaluate the effects of sevoflurane on the expression of early growth response gene 1 (Egr-1) and Egr-2 in the suprachiasmatic nuclei (SCN) of sleep-deprived rats.Methods Forty-eight pathogen-free healthy male Sprague-Dawley rats,were divided into 4 groups (n =12 each) using a random number table:control group (group C),sleep deprivation group (group SD),sevoflurane group (group SEV) and sleep deprivation plus sevoflurane group (group SD+SEV).The rats were subjected to sleep deprivation for 96 h in group SD.The rats inhaled 2.5％ sevoflurane for 3 h in group SEV.The rats inhaled 2.5％ sevoflurane for 3 h after being subjected to sleep deprivation for 96 h in group SD+SEV.The mechanical paw withdrawal threshold (MWT) was measured at 12 h,3 days and 7 days after emergence from anesthesia (T1-3).The animals were sacrificed after blood samples were obtained from the external carotid artery,and the cerebral SCN was removed for determination of Egr-1 and Egr-2 expression by Western blot.Results Compared with group C,the MWT was significantly decreased at T1-3 in SD,SEV and SD+SEV groups,the expression of Egr-1 and Egr-2 in SCN was significantly up-regulated at T1-3 in SD and SD+SEV groups,and the expression of Egr-1 in SCN was significantly up-regulated at T1-3,and the expression of Egr-2 in SCN was up-regulated at T1 in group SEV (P＜0.05).Compared with SD and SEV groups,the MWT was significantly decreased at T1-3 in group SD+SEV,and the expression of Egr-1 and Egr-2 in SCN was significantly up-regulated at T1-3 in group SD+SEV (P＜0.05).Conclusion The mechanism by which sevoflurane aggravates hyperalgesia is related to up-regulation of Egr-1 and Egr-2 expression in SCN of sleepdeprived rats.