Objective To investigate difference of clinical curative effect between laparoscope combined with choledochoscope and traditional laparotomy in treatment of cholecystolithiasis and choledocholithiasis.Methods The clinical data of 80 patients with cholecystolithiasis and choledocholithiasis were retrospectively analyzed,among whom 48 patients were given laparoscopic cholecystectomy (LC) ± laparoscopic common bile duct exploration (LCBDE) (observation group),and 32 patients were given traditional laparotomy (control group).The curative effect was compared between 2 groups.Results There were no statistical differences in operating time,calculi clearance rate and cost of hospitalization between 2 groups (P ＞ 0.05).The intraoperative bleeding,postoperative length of hospital stay,and rate of postoperative complication in observation group were significantly lower than those in control group:(28 ± 5) ml vs.(32 ± 11) ml,(6.0 ± 1.1) d vs.(7.0 ± 1.2) d,4.17％ (2/48) vs.18.75％ (6/32),and there were statistical differences (P ＜ 0.05).Conclusion Laparoscope combined with choledochoscope is an effective treatment method of cholecystolithiasis and choledocholithiasis.

The study of quality control of Chinese medicine has always been the hot and the difficulty spot of the development of traditional Chinese medicine (TCM), which is also one of the key problems restricting the modernization and internationalization of Chinese medicine. Multivariate statistical analysis is an analytical method which is suitable for the analysis of characteristics of TCM. It has been used widely in the study of quality control of TCM. Multivariate Statistical analysis was used for multivariate indicators and variables that appeared in the study of quality control and had certain correlation between each other, to find out the hidden law or the relationship between the data can be found,.which could apply to serve the decision-making and realize the effective quality evaluation of TCM. In this paper, the application of multivariate statistical analysis in the quality control of Chinese medicine was summarized, which could provided the basis for its further study.
Medicine, Chinese Traditional ; standards ; Multivariate Analysis ; Quality Control

AIM:To investigate the effect of pretreatment of stem cell factor(SCF) and granulocyte colony-stimulating factor(G-CSF) on the proliferation and the differentiation of mesenchymal stem cells(MSCs) into cardiomyogenic cells.METHODS:The MSCs,isolated primarily from bone marrow,and purified by passage culture,were obtained from the adult rats of four groups:the rats were pretreated by 5 daily injections of SCF;the rats were pretreated with G-CSF;the rats were pretreated with SCF and G-CSF;the rats were treated without any intervention.The 4th passage of MSCs was labeled by DAPI and cellular cycle analysis was conducted by flow cytometry before co-culture.The neonatal rat cardiomyocytes cultured for 3 days were co-cultured with DAPI-MSCs.The percentage of the differentiation of MSCs into cardiomyogenic cells during the five co-culture days was analyzed.The morphologic changes of MSCs and the proteins expression of cardiac myosin heavy chain(MHC) and troponin T(TnT) were recorded respectively with digital microscope camera system and immunofluorescence technique.The percentage of the differentiation of MSCs into cardiomyogenic cells was also calculated.RESULTS:The percentage of MSCs in G0/G1 phase in SCF/G-CSF group was significantly lower than that in SCF group,G-CSF group and the control group.The percentage of MHC protein-positive MSCs in SCF/G-CSF group was markedly higher than that in SCF group,G-CSF group and the control group,and that in SCF group and G-CSF group was significantly higher than control group.The percentage of TnT protein-positive MSCs in SCF/G-CSF group,SCF group and G-CSF group was significantly higher than that in control group.CONCLUSION:SCF and G-CSF show the ability to stimulate the proliferation of MSCs and induce MSCs to differentiate into cardiomyocytes.The combination of using SCF and G-CSF is more effective than using only SCF or G-CSF.

BACKGROUND: Mesenchymal stem cells (MSCs) can differentiate into myocardial cells in vitro and in vivo, but the amount is small and directed differentiation rate is low.OBJECTIVE: To explore the effect of stem cell factor (SCF) on promotion of MSCs differentiating into myocardial cells.DESIGN: Opening experiment.SETTING: Institute of Cardiovascular Disease, Xianning College and Department of Cardiology, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiment was performed at the Experimental Center of Institute of Cardiovascular Disease, Xianning College from October 2003to August 2004. A total of 20 infant SD rats aged 1-2 days were selected for culture of myocardial cells. Another 25 clean adult SD rats were selected and randomly divided into SCF group, blank control group with 12 rats in each group. The left one rat was used for MSCs extraction, culture and purification.METHODS: ①MSCs were labeled with 4', 6-diamidino-2-phenylindole,dihydrochloride (DAPI) before co-culture. SCF was given into rats of the SCF group by subcutaneous injection successively for 5 days, 20 μg/kg per day. Isolated MSCs were co-cultured with myocardial cells that had been cultured for 3 days. Saline of the same volume was given in the blank control group by subcutaneous injection. MSCs without any intervention were co-cultured with myocardial cells that had been cultured for 3 days.②Expressions of MHC α/β and troponin T were recorded and measured with digital micro-camera shot and immunofluorescence technique, respectively. Percentage of DAPI labeled MSCs differentiating into myocardial cells was measured.MAIN OUTCOME MEASURES: ①Growth of MSCs, ②detection of labeling rate of DAPI on MSCs, ③analysis of activity and purity of co-cultured myocardial cells, and ④differentiation of MSCs into myocardial cells after co-culture.RESULTS: ①Bone marrow cell suspension was inoculated in plastic petri dish. Round cells scattered at the bottom of bottle. At hour 24 a few long fusiform shape adhered cells appeared following liquor change, and at day 4 many fusiform shape adhered cells appeared, which entered logarithm increased period. At days 8-12 80% were confluence. The proliferation of passage cells was more rapid. With liquor change and passage, the MSCs were purified gradually. ②Labeling rate of DAPI on MSCs was 100%. ③At 3-day in vitro culture of myocardial cells the percentage of beat cells was 63%, with beat frequency of 40-60 times per minute. Percentage of positive cells of troponin T expression was 75% examined with immunocytochemical technique. ④At co-cultured days 2 and 3, positive percentage of DAPI labeled MSCs expressing MHC α/β was significantly higher in the SCF group than in the blank control group (P ＜ 0.01). At the co-cultured days 3, 4 and 5, positive percentage of DAPI labeled MSCs expressing troponin T was obviously higher in the SCF group than in the blank control group (P ＜ 0.01).CONCLUSION: SCF has markedly accelerating effect on proliferation and differentiation of MSCs.

miRNAs were discovered less than a decade ago, and have emerged as important regulators of gene expression in mammals. A large number of miRNAs have been identified to be located within the intronic regions of protein-encoding genes(host genes) and called intronic miRNAs. The intronic miRNAs may play a key role in regulating the expression and function of their host genes due to the fact that most of them are co-expressed with the host genes. In this paper, the recent advances on the research on potential relationship between intronic miRNAs and their host genes are reviewed.

Objective To explore the roles of reactive oxygen species(ROS) and TGF-?1 in aldosterone-induced PAI-1 production.Methods Quiescent rat mesangial cells (MCs) were treated by aldosterone.The level of ROS in MCs induced by aldosterone was measured by confecal laser scanning microscopy and the TGF-?1 activity in the supematant of culture was measured by mink lung epithelial cell (Mvllu) proliferation inhibition MTT assay.Then,before the addition of aldosterone,MCs were pretreated with NAC or TGF-?1 neutralizing antibody to decrease cellular ROS or inhibit activity of TGF-?1 induced by aldosterone respectively.PAI-1 mRNA was examined by semi-quantification RT-PCR and PAI-1 protein by Western blotting.Results The intracellular ROS induced by aldosterone increased by 5-fold compared to that of control group,and the activity of TGF-?1 stimulated by aldosterone increased markedly.TGF-?1 neutralizing antibody and NAC effectively decreased aldosterone-induced PAI-1 mRNA expression by 30% and 32%,and PAI-1 protein expression by 21% and 11%,respectively.However,neither TGF-?1 neutralizing antibody nor NAC alone could regulate aldosterone-induced PAI-1 mRNA and protein expression to normal level in 24 hours.Conclusions ROS and TGF-?1 play important roles in up-regulation of aldosterone- induced PAI-1 in MCs.ROS and TGF-?1 are not the exclusive pathway of PAI-1 expression induced by aldosterone in MCs.

ObjectiveTo study the effect of fluorine on proliferation of osteoblast through extra cellular signal-regulated protein kinase(ERK) signaling pathway.MethodsMouse osteoblasts(MC3T3-E1) were cultured in vitro with different concentrations of fluoride for 24 and 48 h (the concentrations of Fˉ were 0,200,400,600,1000,2000,4000,8000,10 000 μmol/L,respectively).The optimum concentration for promotion of cell proliferation was determined by methylthiophene tetrazolium(MTT) assay.According to the optimum concentration,the cells were randomly divided into three groups:control group (0 μmol/L Fˉ); fluorine group (400 μmol/L Fˉ); fluorine and MAPK inhibitor PD98059 group(400 μ mol/L Fˉ + 10 μ mmol/L PD98059).Cell cycle was detected by flow cytometry after 48 h culture.The expression of P-ERK protein was determined by Western blotting and immunofluorescence.ResultsThe optimum concentration of fluorine for proliferation of osteoblasts was 400 μ mol/L.Compared with the control group[(76.12 ± 10.08)％,(2.06 ± 0.31)％],the number of cells in G0/G1 phase[(63.04 ± 8.12)％] reduced and the number of cells in S phase[(9.13 ± 2.08)％] increased in fluorine group (all P ＜ 0.05) ; but the number of cells in G0/G1 phase [(92.11 ± 9.01 ) ％] in fluorine and mitogen-activated protein kinases (MAPK) inhibitor PD98059 group was significantly increased(P ＜ 0.05 ).Western blotting results showed that:compared with the control group[(100.00 ± 0.00)％],the expression of P-ERK protein in fluorine group[(131.24 ± 13.88)％] was significantly higher(P ＜ 0.05 ),but the expression of P-ERK protein in fluorine and MAPK inhibitor PD98059 group [(91.33 ± 9.68 )％] was not significantly changed(P ＞ 0.05).The results of immunofluorescence were similar to that of Western blotting.ConclusionsFluorine at the concentration of 400 μmol/L can promote the proliferation of osteoblasts.ERK signaling pathway has played a key role in the proliferation of osteoblasts.

OBJECTIVETo observe anti-cancer effects of Jianpi Jiedu Recipe (JJR) on liver cancer (LC) rats with Pi deficiency syndrome (PDS) and its relation with the third complementary-determining region gene spectratyping of TCRVβ-chain (TCRVβCDR3).

METHODSRats were divided into 8 groups according to random digit table, i.e., the blank control group (normal), the PDS group, the LC model group, the LC-PDS group, high, middle, and low dose JJR groups (75.00, 37.50, 18.75 g/kg, respectively by gastrogavage, once per day), the thymus pentapeptide group (5 mg/kg, intramuscular injection, twice per week), 8 in each group. Rats in the normal group were administered with physiological saline by gastrogavage once per day. PDS rat model was prepared by bitter-cold purgation. LC model was prepared by orthotopic transplantation method. Twenty gene subfamilies of TCRβCDR3 in the thymus, liver, and LC tissues were detected by Gene Scan.

RESULTSHigh and middle dose JJR could postpone the growth of LC volume (P < 0.05), with equivalent liver index and thymus index to those of the normal group (P > 0.05). In thymus and liver tissue of the normal group, the number of clones (20 and 19), gene fragment number (220 and 113), Quasi-Gaussian distribution ratio of TCRVβCDR3 gene repertoire (100.0% and 42.1%), and fragment fluorescence peak area (6,539 ± 2,325 and 1,238 ± 439) were at the highest level among the 8 groups. TCRVβCDR3 expressions in thymus and liver tissue of high and middle dose JJR groups were approximate to those of the normal group. They were in the middle of the thymus pentapeptide group, the PDS group, the LC model group, and poorest in the LC-PDS group. TCRVβCDR3 in liver tissue expressed the best in the thymus pentapeptide group.

CONCLUSIONJJR might inhibit the growth of LC cells, and its mechanism might be related to enhancing TCRVβCDR3 spectratype expression.

Animals ; Complementarity Determining Regions ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Neoplastic ; Genes, T-Cell Receptor beta ; Liver Neoplasms ; drug therapy ; genetics ; Random Allocation ; Rats

Recombinant human BMP-2 was compounded with chitosan/gelatin/hydroxyapatite(HCG) scaffold and the complex was sterilized by 60Co radiating. Osteoblast isolated from cranial bones of newborn rat was primary cultured and seeded onto the complexes. 3 days after culturing, scanning electron microscope(SEM) was applied to detect the compatibility of the cell with the complex. SEM showed osteoblast attached closely with the complex and grew well in its pores. Then the complexes with osteoblast modification were implanted into athymic nude mice subcutaneously. 8 weeks after implantation, X-ray photograph and histological observation were applied to detect the bone formation of the complexes. Under X-ray a high-density areas consistent with the shape of the implanted complex could be seen. Histological observation also proved there was bone formation in the interspace of the complex. A conclusion was drawn that rhBMP-2 compounded HCG scaffold had good osteogenesis ability in vivo.

OBJECTIVE: To investigate the influence and mechanism of acupuncture at the points in Heel Vessel for the circadian clock genes of Period (Per) 1 and Per 2 mRNAs in the suprachiasmatic nucleus (SCN) in insomnia rats. METHODS: Thirty male SD rats were randomly divided into blank, model, acupuncture groups, 10 rats in each group. Insomnia model was established by intraperitoneal injection of PCPA (suspension, 1 mL/100 g). Acupuncture at "Shenmai" (BL 62) and "Zhaohai" (KI 6) was used in the acupuncture group for continuous 7 days, 15 min/day and once daily. The circadian rhythm was observed; the expressions of Per 1 and Per 2 mRNAs in SCN were examined with real time-PCR. RESULTS: The activity in the model group in rest period everyday increased compared with that in the blank group, and the expressions of Per 1 and Per 2 mRNAs in SCN decreased (P<0.05). Compared with the model group, the activity in the acupuncture group in rest period decreased and the expressions of Per 1 and Per 2 mRNAs in the SCN increased (P<0.05). CONCLUSIONS: Acupuncture at BL 62 and KI 6 can increase the expressions of Per 1 and Per 2 mRNAs in the SCN, so as to decrease the activity in rest period, and improve the quality of sleep in insomnia rats.