To furtherly reduce the subxiphoid port site pain,improve the cosmetic result and patient satisfaction,and increase the safety for patients underwent laparoscopic cholecystectomy by advanced laparoscopic knotting skill.Methods:Among our 1500 patients underwent laparoscopic cholecystectomy since 1991,120 cases of modified laparoscopic cholecystectomy (MLC) were performed with three 5-mm ports and one 10-mm port(for laparoscope and sepcien withdrawn).There were 25 male and 95 female patients with an average age of 55 years (24～77years).The indications for MLC included polypoid lesions of gallbladder (21),simple cholecystitis(3),cholecystolithiasisi with chronic cholecystitis(84),with acute suppurative cholecystitis(7),with atrophic cholecystitis(5).Results:There were 5 patients underwent combined laparoscopic appendectomy(3),fenestration of hepatic cyst(1),and drainge for liver abscess(1).The average operative time for MLC was 55 minutes(30～150min),blood loss was 10ml(3～50ml),and postoperative stay was 3 days(1～5days).There were no conversion from MLC to either LC or open surgery,without mortality.Complications were limited to two patients(1.7%).One was retained common bile duct stone and another was port site bleeding after operation.They were treated by transduodenal endoscopic stone retrieval and simple suture ligation,respecrtively.Conclusions:The advantages of MLC conducted mainly by advanced laparoscopic knotting techniques were no more laparoscope (either 2-mm or 5-mm)needed,no sacrifice of good illumination and laproscopic image.Most of all,its costeffective and operative safety were all improved furtherly.

Objective To evaluate the effects of dexmedetomidine on the activity of cAMP response element binding protein (CREB) and c-fos in the spinal dorsal horn in a rat model of neuropathic pain.Methods Fifty-four adult male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly divided into 3 groups (n =18 each):sham operation group (group S),chronic neuropathic pain group (group C) and dexmedetomidine group (group D).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 350 mg/kg.The sciatic nerve was exposed and 4 ligatures were placed on the right sciatic nerve at 1 mm intervals with 4-0 silk thread in C and D groups.In group D,dexmedetomidine 50 μg/kg was injected intraperitoneally once a day starting from the end of operation until 1 day before the animals were sacrificed,while the equal volme of normal saline was injected instead of dexmedetomidine in S and C groups.Paw withdrawal threshold to mechanical stimulation with yon Frey filament (MWT) and paw withdrawal latency to thermal stimulation (TWL) were measured on 1 day before operation and 3,7 and 14 days after operation.The animals were sacrificed after measurement of MWT and TWL.Their lumbar segments (L4-6) of the spinal cord were removed for measurement of the expression of phosphorylated CREB (pCREB) and c-fos by immunohistochemistry.Results Compared with group S,MWT was significantly decreased,TWL was shortened,and the expression of pCREB and c-fos was up-regulated on 3,7 and 14 days after operation in C and D groups (P ＜ 0.05).Compared with group C,MWT was significantly increased,TWL was prolonged,and the expression of pCREB and c-fos was down-regulated on 3,7 and 14 days after operation in group D (P ＜ 0.05).MWT was significantly lower,and TWL was shorter on 3,7 and 14 days after operation than on 1 day before operation in C and D groups (P ＜ 0.05).MWT was significantly lower,TWL was shorter,and the expression of pCREB and c-fos was higher on 7 and 14 days after operation than on 3 days after operation in C and D groups (P ＜ 0.05).MWT was significantly higher,TWL was longer,and the expression of pCREB and c-fos was lower on 14 days after operation than on 7 days after operation in C and D groups (P ＜ 0.05).Conclusion The mechanism by which dexmedetomidine reduces neuropathic pain is related to inhibition of the activity of CREB and c-fos in the spinal dorsal horn of rats.

Objective To explore how to isolate and culture human primordial germ cells(PGCs).Methods Human embryonic fibroblasts(HEFs) isolated from embryos at 2 to 3 months of age received a radiation of 60Co ?-ray and then served as feeder layer cells.Cells collected from trypsinized human embryonic gonadal ridges and dorsal mesenteries(5 to 9 weeks post-fertilization) were cultured on HEF feeder layers in the medium with recombinant human leukemia inhibitory factor(LIF),recombinant human basic fibroblast growth factor(bFGF) and forskolin.Histochemical and immunocytochemical assays were used to identify the stage specific embryonic antigen-1and 4(SSEA-1/4) of the cultured cells.Reverse transcription polymerase chain reaction(RT-PCR) was used to identify the expressions of specific genes,including Oct-4,Ifitm-3,stella,Mvh and DAZL.Embryonic bodies were cultured as while.Results The collected cells were growing on the feeder layer and formed a typical nestlike multicellular colonies.The cells showed diploid chromosome in karyotype analysis and expressed SSEA-1/4 and specific genes,Oct-4,Ifitm-3,stella,Mvh and DAZL,indicating that the cells were PGCs.When cultured in hanging drop,the PGCs developed into embryonic bodies,showing multipotential ability.Conclusion Human PGCs can be isolated from genital ridges and dorsal mesenteries of embryos and cultured in vitro on HEF feeder layer.The formation of the embryonic stem cell colony can be observed,and the cells cultured by this method is confirmed to be embryonic germ cells.

Objective: To observe the expression of urokinase-type plasminogen activator receptor (uPAR) in human tongue squamous cell carcinoma Tca 8113 cells at different dosages of heat shock. Methods:The expression of uPAR protein in Tca 8113 cells was examined using immunohistochemical technique (IH) and flow cytometry (FCM) after the cells had been treated at 37,40,43 or 45 ℃ for 40 min. Results:The fluorescent intensity of uPAR protein of Tca 8113 cells treated at 37,40,43 and 45 ℃ was 11.36?0.06,9.98?0.12, 10.37?0.05 and 11.31?0.10 respectively.Conclusion:Heat shock can reduce the expression of the uPAR protein of Tca 8113 cells.

Objective To study the cardiac function effect of allopurinol for hypemricemia combined with dilated cardiomyopathy patients.Methods One hundred and twenty hypemricemia combined with dilated cardiomyopathy patients were divided into allopurinol group and control group according to the treatment method with 60 cases each.All the patients were given conventional treatment,the control group was added the nitroprusside treatment,and the allopurinol group was added the allopurinol and nitroprusside treatment.The treatment period was 3 months.Results The total effective rate in allopurinol group was significantly higher than that in control group [90.0％ (54/60) vs.75.0％ (45/60)],there was statistical difference (P ＜ 0.05).After treatment,the left ventricular ejection fraction and left ventricular posterior wall thickness at end-systole in allopurinol group were significantly higher than those in control group [(67.85 ± 7.12)％ vs.(30.78 ±7.00)％ and (1.40 ±0.20) mm vs.(1.16 ±0.18) mm[,but the left ventricular internal diameter at end-systole,left ventricular internal diameter at end-diastole and left ventricular posterior wall thickness at end-diastole were significantly lower than those in control group [(4.72 ± 0.41) mm vs.(6.48 ± 0.47) mm,(2.93 ± 0.32) mm vs.(5.56 ± 0.62) mm and (0.77 ± 0.13) mm vs.(0.92 ± 0.18) mm],there were statistical differences (P ＜ 0.05).After treatment,The uric acid,urea nitrogen and creatinine were significantly lower than those in control group [(45.43 ± 11.24) μ mol/L vs.(167.23 ± 19.22) μ mol/L,(10.23 ± 7.12)mmol/L vs.(40.93 ± 8.09)mmol/L and (32.01 ± 8.34) mmol/L vs.(78.09 ±9.11) mmol/L],there were statistical differences (P ＜ 0.05).Conclusion Allopurinol used for treating hyperuricemia combined with dilated cardiomyopathy patients can reduce uric acid,early reversal the atherosclerosis and improve heart function,it should be widely applied research.

Objective To evaluate the relationship of metabolic syndrome (MS) and MS components with thyroid nodule. Methods A total of 10 357 subjects ( age ＞ 18 years old) who received physical checkup at the Second Affiliated Hospital of Dalian Medical University between June 2009 and June 2010 were enrolled in this retrospective study. Anthropometric parameter, fasting plasma glucose (FPG),serum lipid profile, blood uric acid, alanine aminotransferase and thyroid ultrasonography were measured. Results The prevalence of thyroid nodule,MS,and thyroid nodule + MS was 46. 96% ,23. 6%,and 11.6%, respectively. The prevalence of thyroid nodule was significantly higher in MS patients than in non MS patients ( 75.9% vs 38. 0%, P ＜ 0. 05 ). Multifactor logistic analysis showed that MS, body mass index (BMI) and FBG (β vales were 0. 78,1.22,and 0. 62,respectively; odds ratios were 4. 167,3. 876,and 2. 359, respectively; all P ＜ 0. 05 ) were independently correlated with the development of thyroid nodule. Conclusions Significantly increased prevalence of thyroid nodule could be found in MS patients. BMI and FBG may be independent risk factors of thyroid nodule.

Objective To evaluate the effect of dexmedetornidine on the expression of Toll-like receptor 4 (TLR4) and nuclear factor kappa B (NF-κB) in the spinal cord in rats with neuropathic pain (NP).Methods One hundred and eight male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly assigned into 3 groups (n =36 each):sham operation group (group S),NP group and dexmedetomidine group (group D).NP was induced by chronic constrictive injury in anesthetized rats.Sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals with 4-0 silk thread.In group S,the right sciatic nerves were exposed,but not ligated.Dexmedetomidine 50 μg/kg was injected intraperitoneally once a day from the onset of operation to one day before the rats were sacrificed in group D,while the equal volume of normal saline was injected in groups S and NP.Mechanical withdrawal threshold (MWT) and thermal pain threshold (TPT) were measured on the day before operation (T0) and 3,7,and 14 days after operation (T1-3).After measurement of pain threshold at T1,T2 and T3 after operation,the L4-6 segments of the spinal cord were removed for determination of the expres-sion of TLR4 and NF-κB mRNA (by RT-PCR) and the expression of TLR4 and NF-κB in spinal dorsal horn (by immuno-histochemistry).Results Compared with group S,MWT and TPT were significantly decreased and the expression of TLR4,NF-κB and TLR4 and NF-κB mRNA was up-regulated after operation in groups NP and D (P ＜ 0.05).Compared with group NP,TPT and MWT were significantly increased and the expression of TLR4,NF-κB,TLR4 mRNA and NF-κB mRNA was significantly down-regulated after operation in group D (P ＜ 0.05).Conclusion The mechanism by which dexmedetomidine attenuates NP in rats is related to inhibition of the expression of TLR4 and NF-κB in rat spinal cord.

Objective To evaluate the effect of dexmedetomidine and small dose of ketamine on the expression of P2X4 receptor (P2X4 R) mRNA and P2X7 receptor (P2X7R) mRNA in the dorsal root ganglion of rats with neuropathic pain.Methods Ninety male Sprague-Dawley rats,aged 6-9 weeks,weighing 180-220 g,were randomly divided into 5 groups (n =18 each):sham group (group S),chronic constrictive injury group (group CCI),dexmedetomidine group (group D),ketamine group (group K) and dexmedetomidine + ketamine group (group DK).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 400 mg/kg.Neuropathic pain was induced by CCI in CCI,D,K and DK groups.The sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1mmintervals with 4-0 silk thread.In group S,the sciatic nerves were only exposed but not ligated.In D,K and DK groups,dexmedetomidine 50μg/kg,ketamine 10 mg/kg and dexmedetomidine 25μg/kg + ketamine 5 mg/kg were injected intraperitoneally,respectively,while the equal volume of normal saline was injected in S and CCI groups,once a day for 14 consecutive days after CCI.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before CCI,and 3,7 and 14 days after CCI.Six animals were sacrificed after measurement of pain threshold at 3,7 and 14 days after CCI and the lumbar segments (L4-6) of the dorsal root ganglion were removed for determination of P2X4 R mRNA and P2X7 R mRNA expression by RT-PCR.Results Compared with group S,MWT and TWL were significantly decreased at 3,7 and 14 days after CCI in groups CCI,D,K and DK,the expression of P2X4R mRNA and P2X7R mRNA was up-regulated at 3,7 and 14 days after CCI in groups CCI,D and K,and the expression of P2X4 R mRNA and P2X7 R mRNA was up-regulated at 3 and 7 days after CCI in group DK (P ＜ 0.05).Compared with group CCI,TWL and MWT were significantly increased and the expression of P2X4 R mRNA and P2X7 R mRNA was down-regulated at 3,7 and 14 days after CCI in groups D,K and DK (P ＜ 0.05).Compared with D and K groups,TWL and MWT were significantly increased and the expression of P2X4 R mRNA and P2X7 R mRNA was down-regulated at 3,7 and 14 days after CCI in group DK (P ＜ 0.05).Conclusion The mechanism by which the combination of dexmedetomidine and small dose of ketamine produces a synergistic antinociception in rats with neuropathic pain may be related to down-regulation of the expression of P2X4 R mRNA and P2X7 R mRNA.

Objective To investigate the effects of Sulfotanshinone Sodium Injection (SSI) on neuropathic pain in rats.Methods One hundred and eight adult male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly divided into 3 groups (n =36 each):sham operation group (group S) ; chronic constrictive injury (CCI)group; group SSI.The animals were anesthetized with intraperitoneal 10% chloral hydrate 350 mg/kg.In groups CCI and SSI,4 ligatures were placed on the right sciatic nerve at 1 mm intervals with 4-0 silk thread according to the method described by Bennett et al.In group S,the right sciatic nerves were exposed,but not ligated.In group SSI,SSI 25 mg/kg was injected intraperitoneally once a day starting from the end of operation until one day before the animals were sacrificed,while the rats received the equal volume of normal saline (5 ml/kg) instead of SSI in groups S and CCI.Twelve animals in each group were chosen at 1 day before operation and 3,7 and 14 days after CCI (T1-4) to measure mechanical paw withdrawal threshold to yon Frey stimuli (MWT) and paw withdrawal latency to thermal nociceptive stimulus (TWL).Six rats in each group were sacrificed at T2-4 after measurement of pain threshold,and their lumbar segnents (L4-6) of the spinal cord were immediately removed for determination of Bcl2 and caspase-3 expression in spinal dorsal horn (by immune-histochemistry),and MDA content and SOD activity (by spectrophotometry) in spinal cord.Results Compared with group S,PWT was significantly decreased,PWL was shortened,the expression of Bcl-2 and caspase-3 was up-regulated,MDA content was increased and SOD activity was decreased at T2-4 in groups CCI and SSI (P ＜ 0.05).Compared with group CNP,PWT was significantly increased,PWL was prolonged,the expression of Bcl-2 was up-regulated,the expression of caspase-3 was downregulated,MDA content was decreased and SOD activity was increased at T2-4 in group SSI (P ＜ 0.05).Conclusion SSI can mitigate neuropathic pain in rats and inhibition of oxidative stress in spinal cord tissues and reduction of apoptosis in spinal dorsal horn neurons are involved in the mechanism.

ObjectiveTo evaluate the therapeutic effect ofZishen decoction combined with standard treatment of western medicine for for stage IV diabetic nephropathy with Qi and Yin deficiency.MethodsA total of 112 patients with stage IV diabetic nephropathy and Qi and Yin deficiency were randomized to thestandard treatment and the combined treatment groups, 56 in each. The standard treatment group received conventional treatment, including blood glucose controlling, antihypertensive, blood lipid regulating and diet controlling. The combined treatment group receivedZishen decoction on the basis of conventional treatment. All the patients were treated for 3 months. The blood urea nitrogen (BUN), serum creatinine (SCr), total cholesterol (TG), triacylglycerol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and glycated hemoglobin (HbA1c) were measured by an automatic chemistry analyzer. The urinary albumin excretion rate (UAER) was measured by the enzyme-linked immunosorbent assay.ResultsCompared with the standard treatment group, the SCr (53.51 ± 18.12μmol/Lvs. 62.66 ± 21.14μmol/L;t=2.459,P<0.05), UAER(100.73±84.24μg/minvs. 156.24 ± 96.38μg/min;t=3.245,P<0.05), TG(1.73±0.22 mmol/Lvs. 2.06 ± 0.21 mmol/L;t=8.112,P<0.01), TC(4.56 ± 0.62 mmol/Lvs. 5.10 ±0.31 mmol/L;t=5.830, P<0.01), LDL-C (2.42 ± 1.05 mmol/Lvs. 3.31 ± 0.81 mmol/L;t=5.022,P<0.01) in the combined treatment group decreased significantly, and the HDL-C (1.67 ± 0.33 mmol/Lvs. 1.36 ± 0.41 mmol/L;t=4.460,P<0.01) increased significantly. ConclusionZishen decoction on the basis of conventional treatment can improve the SCr and UAER, and regulate the blood lipid in the patients with stage IV diabetic nephropathy and Qi-Yin deficiency.