Objective:To establish a method for isolation,purification,and culture of human multipotent adult progenitor cells(MAPCs) in vitro,so as to lay a foundation for the application of hMAPCs in tissue-engineering research and clinical medicine.Methods: The mononuclear cells were obtained from bone marrow of healthy volunteers by gradient centrifugation and were subjected to adherence culture.The adherent cells were then subjected to magnetic activated cell sorting(MACS)(depletion selection with CD45 and GlyA microbeads).Then the purity of selected cells was identified by flow cytometer.The growth of the purified cells was observed and the expression of CD29,CD44,CD34,and HLA-DR was analyzed by flow cytometers.Results: (1)Averagely(5-10)?10~(4)/ml hMAPCs could be separated from 1?10~(6)/ml bone marrow mononuclear cells by MACS.The cell viability was similar before(96.7?1.7%) and after(96.0?2.4%) isolation.(2)The isolated MAPCs grew well in the self-designed culture medium and could be passaged for 20 generation.(3)The purity of the CD45~-and GlyA~-cells separated from bone marrow adherent cells was more than 98% as determined by flow cytometer.(4) In hMAPCs,the positive rate was 99.2% for CD29,98.3% for CD44,1.2% for CD34,and 5.3% for HLA-DR.Conclusion:(1)The bone marrow-derived hMAPCs can be purified by MACS through depletion selection of CD45 and GlyA microbeads.(2)The hMAPCs can be expanded in vitro in self-designed medium,maintaining a non-differentiation state for a long time.

Objective: To investigate the protective effect of astilbin on warm ischemia/reperfusion-induced liver injury and to study the related mechanism. Methods: C57BL/6 mice were randomly divided into four groups (n=8): sham-operated group (Sham), model control group (I/R), low dose astilbin treatment group (10 mg/kg) and high dose astilbin (40 mg/kg) treatment group. Mice in the two treatment groups were intraperitoneally injected with astilbin 24 hours and one hour before ischemia. Then 70% hepatic ischemia/reperfusion model (the left and middle hepatic lobe) was established. Mice in the I/R model control group and the sham operation group were administered with the same volume of normal saline. The blood sample and liver tissue samples were collected 90 min after ischemia and 6 h after reperfusion. Serum ALT activity was detected as an indicator of liver function damage and the content of MPO in liver tissues were detected by ELISA. The pathological changes of the liver were observed. The expression of IL-10 in liver tissues was detected by Western blotting and the expression of IL- 10 mRNA was detected by semi-quantitative RT-PCR. Results: Astilbin treatment can effectively lower the serum ALT level and MPO level in the liver tissues in some I/R mice. It could also improve the pathological manifestations of the liver. Compared with the I/R model control group, IL-10 protein levels gradually increased in the two treatment groups, which was consistent with the result of RT-PCR (low dose group P<0.05; high dose group, P<0.01). Conclusion: Astilbin can effectively reduce the inflammatory response after liver warm ischemia-reperfusion induced injury, effectively improve the mouse liver function and pathological damage, which might be related to the upregulation of IL-10 expression in the liver tissues.

Objective: To establish a method for isolation, purification, and culture of human multipotent adult progenitor cells (MAPCs) in vitro, so as to lay a foundation for the application of hMAPCs in tissue-engineering research and clinical medicine. Methods: The mononuclear cells were obtained from bone marrow of healthy volunteers by gradient centrifugation and were subjected to adherence culture. The adherent cells were then subjected to magnetic activated cell sorting (MACS) (depletion selection with CD45 and GlyA microbeads). Then the purity of selected cells was identified by flow cytometer. The growth of the purified cells was observed and the expression of CD29, CD44, CD34, and HLA-DR was analyzed by flow cytometers. Results: (1) Averagely (5-10) × 104/ml hMAPCs could be separated from 1 × 106/ml bone marrow mononuclear cells by MACS. The cell viability was similar before ([96.7±1.7]%) and after ([96.0±2.4]%) isolation. (2) The isolated MAPCs grew well in the self-designed culture medium and could be passaged for 20 generation. (3) The purity of the CD45- and GlyA- cells separated from bone marrow adherent cells was more than 98% as determined by flow cytometer. (4) In hMAPCs, the positive rate was 99.2% for CD29, 98.3% for CD44, 1.2% for CD34, and 5.3% for HLA-DR. Conclusion: (1) The bone marrow-derived hMAPCs can be purified by MACS through depletion selection of CD45 and GlyA microbeads. (2) The hMAPCs can be expanded in vitro in self-designed medium, maintaining a non-differentiation state for a long time.

ObjectiveThe relationship between clinicopathological features and prognosis in the women patients with breast mucinous carcinoma was investigated. Methods 101 patients with breast mucinous carcinoma were divided into two groups:pure mucinous carcinoma ( n =74), and mixed type ( n =27). The relationship between clinicopathological features and prognosis was analysed. Results Lymph node was less frequently involved in pure type group (24%) than that in the mixed type (52%) (? 2 =10 4, P

Objective To explore the changes and their correlation of event-related potential(ERP)and the plasma levels of monoamine neurotransmitter in the patients with epilepsy.Methods ERP and the plasma levels of noradrenalin(NE),dopamine(DA) and 5-hydroxytryptamine(5-HT) were tested in 50 patients with epilepsy,and the results were compared with the normal controls.Results The latency of N2,P3 of ERP and the amplitude of P3 in the epilepsy group were significant longer and lower than those in the normal control group(all P

BACKGROUND:. Until now modern science cannot explain clearly aboutthe mechanism of acute alcoholic liver injury. Besides such methods as givingup drinking, or symptomatic treatment, there is no specific therapy and remedy. Traditional Chinese Medicine (TCM) has obvious advantages for this: Itcannot only improve the clinical symptoms, but also adjust organism immunefunction, and has very good development future. OBJECTIVE: To study the effects of radix bupleuri and radix scutellariae incompatibility with the ratio of different dosage (2:1, 1:1, 1:2) on acute alcoholicliver injury by regulating the system of endocrine-immunity network in rats. DESIGN: Groups are divided randomly and experiments are made underblank conditions. SETTING: TCM Clinical Laboratory of Pharmaceutical School, Henan University of TCM. MATERIALS: From September to November 2002 in TCM Clinical Laboratory of Pharmaceutical School, Henan University of TCM, the general condi tion of the animals were as follows: 96 healthy Wistar rats were divided into 8groups: blank group, model group, radix bupleuri group, radix scutellariaegroup, xiao-chaihu group, radix bupleuri-radix scutellariae 2:1 group, radix bupleuri-radix scutellariae 1:1 group, radix bupleuri-radix scutellariae 1:2 group. METHODS: ① According to the ratio of 2:1, 1:1, 1:2, radix bupleuri andradix scutellariae were taken respectively (480 g, 240 g; 360 g, 360 g; 240 g, 480 g) to produce the water decocting nedicine. Except the blank group(duplicated by saline of the same volume through consecutive gastric infusion), the other groups are duplicated through consecutive gastric infusionby 56 degrees of Er-Guotou White Spirit (7 mL/kg) two times a day for tendays until the model-making is finished. From the first day, the medicine wasgiven two times a day. The blank group and the model group were given thedistilled water of the same volume. The other groups were given the properwater decocting medicine. The dosage in xiao-chaihu group was 10 g/kg andin the other was all 12 g/kg. ② 16 hours later after the last time when thewhite spirit was given, blood was taken from stomach aorta to produceserum. Liver, spleen, chest gland, adrenal gland were quickly removed. Thecalculating formula of an organ index: An organ index= The weight of an organ (g)/The weight of a body (g) ×100%. The calculating formula of vitamin C content in adrenal gland (mg/g): Content of vitamin C= [(The Avalue of the testing tube-The A value of the blank tube)/ (The A value ofthe standard tube-The A value of the blank tube)] ×density of the standardtube (6 mg/L)×dilution times of the sample before tested/protein content(g/L). ③ Comparison between the groups was shown by using variancesimilar test and single-factor variance analysis.MAIN OUTCOME MEASURES: Index of rats' liver, spleen, chest gland, and adrenal gland (on the right side); the determination of vitamin C content in rats' right adrenal gland.RESULTS: During the experiment, 3 rats in the model group died. Tworats died in radix scutellariae group and radix bupleuri-radix scutellariae group respectively, In each of the other groups only one rat died. The condition of the number of the rats which were used for analyzing results:There were 12 in blank group; 9 in model group; 10 in radix bupleuri group, radix bupleuri-radix scutellariae 1:1 group respectively; 11 in each of the other groups. ① Comparison of index of rats in liver, spleen, chest gland, adrenal gland (right side) between different groups: There was no significant difference between the indexes of rats' liver in different groups.The index of spleen, chest gland, adrenal gland in the model group were all greatly less than in blank group (P ＜ 0.05-0.01). The index of spleen in the radix bupleuri group wcre greatly higher than in the model group, it could make the reduced spleen index return to normal (P ＜ 0.05); The index of chest in radix scutellariae group, radix bupleuri-radix scutellariae 1:1 group, radix bupleuri-radix scutellariae 1:2 group and xiao-chaihu group were greatly higher than in model group (P ＜ 0.05); The index of adrenal gland in radix bupleuri-radix scutellariae 1:2 group and xiao-chaihu group were greatly higher than in model group (P ＜ 0.05). ② Comparison of vitamin C content in rats' right adrenal gland between the groups: Compared with blank group, the vitamin C content of adrenal gland in the model group had no significant difference. While that of radix bupleuri group, xiao-chaihu group, radix bupleuri-radix scutellariae groups were greatly less than that of model group (P ＜ 0.05-0.01), especially radix bupleuri-radix scutellariae 1:1 group had the most significant influence (P ＜ 0.01).CONCLUSION: Interfering functional injuries of liver, spleen and suprarenal gland is one of the mechanisms of acute alcoholic liver injury.Generally speaking, bupleuri-radix scutellariae 1:2 group has the greatest effect on the testing index.

Objective The objectives of this study were to investigate the clinical significance of CD39,CD73,double positive subgroups for CD39 and CD73,and other lymphocytes with clinicopathological parameters in the microenvironment of colorectal cancer.Methods Tumor infiltrating lymphocyte(TIL)was collected from 24 patients with colorectal cancer after radical resection.The expression of CD39+,CD73+ or CD39+ with CD73+ in T cells were measured by flow cytometry.The association between these subgroups and clinicopathologic parameters was analyzed.Results The CD73+ and CD39+ with CD73+ subgroups were associated with lymph node metastasis and poor degree of differentiation,and this mechanism was closely related to tumor-associated inflammation.Conclusion CD39+ with CD73+ colorectal tumor infiltration Treg has a more unique biological activity than other Treg group.This study provides a new idea and theoretical basis for predicting the prognosis of colorectal cancer.

Background Measurement of ciliary sulcus distance is important for phakic intraocular lens implantation.Suowei panoramic 50-MHz digital ultrasound biomicroscopy makes it possible for the direct measurement of ciliary sulcus distance.Objective The aim of this study was to investigate the correlation between anterior chamber(AC)angle diameter and ciliary sulcus distance in different AC depth population and offer a clinical guide.Methods The anterior chamber angle diameter and ciliary sulcus distance were measured in 30 right eyes with shallow anterior chamber caused by suspect glaucoma and 30 right eyes with deep anterior chamber dut to the high myopia with Suowei panoramic 50-MHz digital ultrasound biomicroscopy.Three horizontal B-scans with the focal plane approximately at a depth corresponding to the iris plane were acquired for each eye.The differences of the anterior chamber angle diameter and ciliary sulcus distance were calculated and compared between the two groups.The correlation of the anterior chamber angle diameter and ciliary sulcus distance were evaluated in respective group.Results In shallow AC group,the mean angle diameter was(11.49+0.75)mm,the mean ciliary sulcus distance was (10.97±0.86)mm with the mean standard deviation of the difference of(-5.14± 0.55)mm,showing a statistically significant difference(t =-5.092,P =0.000).In shallow AC group,linear regression analysis showed a statistically significant correlation between ciliary sulcus distance and angle diameter(R2=0.593,P=0.000)and the Pearson's correlation coefficient was 0.767.In deep AC group,the angle diameter was(12.69±0.67)mm;while ciliary sulcus distance was(12.31 ± 0.61)mm with the mean slandard deviation(-0.38 ± 0.17)mm,which was statistically significantly different(t=-5.531,P=0.000).The ciliary sulcus distance was found to be linear correlation with the angle diameter(R2 =0.699,P=0.000)and Pearson' s correlation coefficient of 0.836.Conclusions Panoramic ultrasound biomicroscopy is an useful tool to the measurement of ciliary sulcus distance.This study exhibited that the ciliary sulcus distance is significant associated with the anterior chamber angle diameter,which may offer a guide for phakic intraocular lens implantation.

Objective To explore the reactive inhibition and electrophysiological changes under the GO/NOGO paradigm in patients with alcohol dependence (AD) in order to provide a theoretical basis for neuromechanism study of AD executive dysfunctions.Methods The GO/NOGO paradigm event related potentials (ERP) were tested in 51 AD patients and 29 healthy controls, this paradigm behavior and ERP-N1, N2 and P3 latency and amplitude were analyzed.ERPs and execution functions were followed at different time points of withdrawal(4, 9,13 weeks).Results The number of correct responses (29.18±3.03)lowered and error responses (5.16±3.77) heightened in AD patients, and differences had statistical significance compared with those in control group(32.35±2.58,3.19± 1.84, P＜0.05), and there was no group difference in response time.There was no statistical significance in group difference in N1 component under the GO/NOGO tasks and in P3 latency under GO.N2 ((3.48±2.21) μV, (4.49±3.00) pV) and P3 amplitude ((3.47±2.31) μV, (3.92±4.09) μV) decreased and N2 latency ((256.30±31.62) ms, (256.09±33.77)ms) prolonged in AD group under both GO/NOGO tasks and so did P3 latency ((355.33±25.96) ms) under the NOGO, and the differences had statistical significance.Group main effects in N2 and P3 component (P=0.000) and task main effects in N2 amplitude and P3 latency (P＜0.05) were significant,group and task interaction was significant only in P3 latency (P＜0.01).ERPs of AD patients had no notable changes at different time points of withdrawal, executive dysfunctions were the worst within the 1st week of withdrawal and gradually recovered to pre-withdrawal levels.Conclusion AD patients have lower reactive inhibition manifested mainly by weakened capacity to inhibit and adjust conflict monitor and pre-executive motor program, ERP and executive functions cannot be well improved during short-term withdrawal, and the GO/NOGO-ERP can completely reflect electrophysiological changes of reactive inhibition process of AD patients.

Objective To investigate the effective strategies to prevent and treat biliary complications after orthotopic liver transplantation.Methods The clinical data of 316 patients who received orthotopic liver transplantation at the Fuzhou General Hospital of Nanjing Military Command from November 2001 to March 2012 were retrospectively analyzed.Cold perfusion with HTK + UW solution was applied when obtaining the liver graft,and then the liver graft was preserved in the UW solution.The bile duct was perfused with UW solution thereafter.Orthotopic liver transplantation or piggyback liver transplantation were adopted in the cadaver liver transplantation.Left liver transplantation and right liver transplantation were adopted in the living donor liver transplantation.Choledochojejunal Roux-en-Y anastomosis or duct-to-duct choledochostomy were used for biliary reconstruction.Ordinary T tubes were used for drainage before 2006,and then 6 F pediatric suction catheter or epidural catheter were applied for drainage thereafter.The Ttube was pulled out 3-6 months after the operation.Enteral nutrition was applied to patients at the early phase after operation.The immunosuppressive agents used including tacrolimus + mycophenolatemofetil + adrenal cortical hormone,and for some patients,tacrolimus + mycophenolatemofetil + sirolimus + hormone were used.Patients were followed up for 2 years to learn the incidence of biliary complications and guide the medication.The difference in the incidence of bile leakage between patients who wcrc admitted before 2006 and those admitted after 2006 were compared using the chi-square test.Results The warm ischemia time was 2-6 minutes,and the cold ischemia time was 3-10 hours.For patients who received cadaver liver transplantation,orthotopic liver transplantation was carried out for 291 times and piggyback liver transplantation for 24 times; biliojejunal Roux-en-Y anastomosis was carried out for 5 times and bile duct end-to-end anastomosis for 310 times.For patients who received living donor liver transplantation,1 received left liver transplantation and 1 received right liver transplantation,and they received bile duct end-to-end anastomosis.A total of 311 patients received immunosuppressive treatment with tacrolimus + mycophenolatemofetil + adrenal cortical hormone,and 5 patients reveived tacrolimus + mycophenolatemofetil + sirolimus + hormone.Of the 316 patients who received orthotopic liver transplantation,38 had biliary complications after the operation,including bile leakage in 18 patients,intra-and extra-hepatic bile duct stricture in 6 patients,anastomotic stricture in 6 patients,biliarycomplications included cholangitis in the portal area and cholestasis in 4 patients,choledocholithiasis and cholangitis in 2 patients and biliary infection in 2 patients.The incidence of bile leakage before 2006 was 14.00％ (7/50),which was significantly higher than 4.12％ (11/267) of bile leakage after 2006 (x2-7.676,P ＜ 0.05).Of the 38 patients with biliary complications,the condition of 35 patients was improved,and 3 patients died.Of the 18 patients with bile leakage,15 was cured by conservative treatment,3 received surgical treatment (the condition of 1 patient was improved by drainage,anti-infection treatment and nutritional support,but died of peritoneal hemorrhage at postoperative 1 month; 2 patients received peritoneal drainage,1 was cured and 1 died of peritoneal infection).For the 6 patients with intra-and extra-hepatic bile duct stricture,1 was cured by liver retransplantation and 5 were cured by conservative treatment,endoscopic retrograde cholangio-pancreatography (ERCP) or balloon dilation.For the 6 patients with anastomotic stricture,the condition of 3 patients was improved by conservative treatment,balloon dilation or stent implantation,1 gave up treatment due to hepatic cancer recurrence and died thereafter,1 received anastomosis + T tube drainage,1 was cured by recurrent tumor resection and choledochojejunostomy.Four patients with cholangitis in the portal area and cholestasis were cured by conservative treatment.For the 2 patients with choledocholithiasis and cholangitis,1 was cured by stent implantation with ERCP,and 1 received conservative treatment,and the level of total bilirubin was decreased.Two patients with biliary infection were cured by anti-infection treatment.Conclusions Most of the biliary complications could be treated by non-surgical treatments.For patients with severe biliary complications or those could not be treated by non-surgical treatment,re-exploration of the bile duct is effective.Liver re-transplantation is the only choice for patients with dysfunction of liver graft caused by severe ischemic biliary injury.