Objective:To compare the clinical efficacy of Du's heat-reinforcing method and Western medication in treating diarrhea-predominant irritable bowel syndrome (IBS-D).Methods:Sixty-five IBS-D patients were randomized into two groups by the random number table.Thirty-three cases in the treatment group were intervened by acupuncture with Du's heat-reinforcing method,while thirty-two cases in the control group were given oral administration of pinaverium bromide tablets.The intervention lasted 4 weeks for both groups.The IBS symptom severity score (IBS-SSS),IBS-quality of life (IBS-QOL) and hospital anxiety and depression scale (HAD) were adopted to evaluate the therapeutic efficacy.Results:The two groups each had two dropout cases.The clinical recovery rate was 38.7％ and the total effective rate was 90.3％ in the treatment group versus 13.3％ and 66.7％ in the control group.The treatment group was superior to the control group in both clinical recovery rate and total effective rate (both P＜0.05).After 1-week and 4-week treatment,respectively,the IBS-SSS scores were lower compared with the baseline in both groups,and the intra-group differences were statistically significant (both P＜0.05);the score in the treatment group was lower than that in the control group,and the between-group difference was statistically significant (both P＜0.05).After 4-week treatment,the component scores of IBS-QOL showed improvements in both groups,and the treatment group was superior to the control group in the improvements of dysphoria,interference with activity,health worry and food avoidance (all P＜0.05).The anxiety and depression scales of HAD (HAD-a,HAD-d) in the treatment group and the HAD-a in the control group obtained significant improvements (all P＜0.05);the scores of HAD-a and HAD-d in the treatment group were significantly better than those in the control group (both P＜0.05).Conclusion:Acupuncture with Du's heat-reinforcing method can effectively ease the symptoms of IBS-D,improve the quality of life and the state of anxiety and depression.It can produce a more significant efficacy than oral administration of pinaverium bromide tablets.

Objective To investigate the distribution of ompA gnne in 15 Chinese reference standard strains belonging to 15 serogroups of Leptospira interrogate, and to express recombinant OmpA ( rOmpA ) and to identify immunogenicity and immunoprotection of rOmpA. Methods Genomic DNAs from different leptospiral strains were extracted by phenol-chloroform method. Entire ompA gene fragments from the strains were amplified by PCR and then sequenced after T-A cloning. A prokaryotic expression system of ompA gene from L. interrogans strain 56601 was constructed, and the expression and yield of rOmpA were determined by SOS-PAGE plus Bio-Rad Agarose Image Analyser. Rabbits were immunized with rOmpA for obtaining antiserum, and immunodiffusion test was used to measure the antiserum's titer. Western blot assay was performed to determine the immunoreaetivity of rOmpA with the antiserum against rOmpA and antiserum against whole cell of L. interrogans strain 56601, while mi-croscopic agglutination test (MAT) was applied to detect the cross agglutination to the 15 L. interrogans strains. A leptospire adhering cell model and a leptospire infecting guinea pigs model were used to determine the adhesion-bloc-king effect of rOmpA antiserum and immunoprotection of rOmpA. Results All the 15 L. interrogans strains, but not L. biflexa strain Patoe Ⅰ , had sequence conserved ompA genes. The yield of rOmpA was approximate 20% of the total bacterial proteins, rOmpA could induce rabbits to produce antibody and immunodiffusion titer of the anti-serum was 1:4. Both antisera against rOmpA and against whole cell of L. interrogans strain 56601 were able to pro-duce positive Western blot signs to rOmpA, and the former offered 1 : 20-1 : 320 MAT titers to the 15 L. interrogans strains. 1: 10-1:160 dilutions of rOmpA antiserum could efficiently block L. interrogans strain 56601 adhering to J774A. 1 cells, and 100 μg and 200 μg rOmpA displayed 50.0% and 75.0% immunoprotective rates in the infee-ted guinea pigs. Conclusion ompA gene only exists in genomes of different pathogenic L. interrogans serogroups. rOmpA has relatively stronger antigenicity, cross immunoreactivity and certain immunoprotection, implying that this recombinant protein may be used as a candidate antigen for developing universal genetic engineering vaccine of L. interrogans.

Objective:To study the function of transcription factor 4 (TCF4) and to prepare TCF4 polyclonal antibody.Methods:pET-41/TCF4 was transformed into E.coli BL21(DE3) and induced by IPTG.The purified GST-TCF4 fusion protein was applied to immunize rabbit to produce antiserum. The specificity of the affinity of purified anti-TCF4 antibody was examined by Western blotting analysis of the eukaryotic expressed products of TCF4. Dig-labeled probe and antibody against TCF4 were used to examine the expression of TCF4 in Saos-2 cells under mechanical stretch. Results:Western blotting showed that the antibody could bind to TCF4 specifically. The expression of TCF4 mRNA and protein were significantly increased in Saos-2 cells under mechanical stretch. Conclusion: TCF4 antibody has been prepared successfully.

Objective:To evaluate the status quo and influencing factors of hyperlipidemia management in patients with contracted family doctor service in the community.Method:The baseline data and blood lipid testing results of 752 hyperlipidemia patients (334 males and 418 females) with contracted family doctor service in Yuetan Community Health Service Center from November?2019 to May 2020 were collected. The hyperlipidemic patients were managed by family doctors based on atherosclerotic cardiovascular diseases(ASCVD) riks assessment. The ASCVD risk levels and low-density lipoprotein cholesterol (LDL-C) compliance rate of patients with different general data were compared, and the influencing factors of LDL-C control failure were analyzed by logistic regression.Results:The ASCVD risk assessment showed that among 752 patients there were 172 cases of low risk(22.87%), 167 cases of moderate risk(22.21%),352 cases of high risk(46.81%) and 61 cases of extremely high risk(8.11%). A significant difference was detected in sex,rate of smoking,incidence of overweight or obesity among patients with different ASCVD risk levels ( P<0.05).The overall control rate of LDL-C was 48.8% (367/752), that for low, moderate, high and extremely high risk patients were 83.73% (144/172), 53.89% (90/167), 34.38% (121/352) and 19.67%(12/61), respectively. A significant difference was detected in sex(female: 52.87%, 221/418),age(aged over 80: 58.82%, 110/187), rate of smoking (non-smoking:52.40%, 327/624) and medication compliance (good compliance:52.87%,221/418) between LDL-C control and uncontrol groups (χ2=6.323,11.816,19.022,25.274; P<0.05). Multiple logistic regression analysis revealed that male gender ( OR=1.800,95% CI:1.325-2.419), smoking ( OR=2.630,95% CI:1.726-4.007) and poor medication compliance ( OR= 2.179, 95% CI: 1.581-3.003) were independent risk factors for uncontrolled LDL-C levels. Conclusion:Patients with hyperlipidemia have a relatively high risk of cardiovascular diseases, and their blood lipids are not well controlled. The management of blood lipid should be enhanced in patients with chronic diseases, particularly for male patients with smoking and poor medication compliance.

Objective To investigate the effect of prostaglandin E: (PGE1) on recovery of early renal graft functions after transplantation. Methods One hundred and seven patients after renal transplantation were allocated in the treated group, and treated by conventional treatment with injection of 10 μg prostaglandin E1 additionally twice a day for 14 days. And eighty-eight patients who received conventional treatment alone after renal transplantation at the corresponding period were allocated in the control group. Indexes of the two groups, including incidence of delayed graft function and acute rejection reaction, volume of urine, serum certaintie (SCr), endogenous certainties clearance rate (CCr), the blood flow resistance in graft as well as blood viscosity (BV), and platelet aggregation rate (PAR), were determined. Results The urinary volume and endogenous certainties clearance rate of the treated group were significantly higher, but the level of SCr, incidence of renal function recovery retardation, BV, PAR and blood flow resistance in graft were significantly lower than these of the control group (P<0.05). The difference of incidence of acute rejection reaction between the two groups was insignificant (P>0.05). Conclusion Prostaglandin E1 can improve blood microcirculation and decrease the incidence of renal function recovery retardation. These effects are helpful for recovery of renal function after renal transplantation.

The present study was undertaken to define whether intracarotid injection of capsaicin induces Fos expression associated with the activation of NOS-containing neurons in brainstem nuclei by combining the immunocytochemical method for Fos with NADPH-d histochemical technique for NOS. The results obtained are as follows: (1) Intracarotid injection of capsaicin caused a significant increase of Fos-like immunoreactive neurons in area postrema (AP), nucleus tractus solitarius (NTS), paragigantocellularis lateralis (PGL) and locus coeruleus (LC), without influence upon the neurons of raphe nuclei (RN) and periaqueductal gray (PAG). (2) NO-containing neurons in PGL and NTS and the double-labeled neurons in PGL were also increased significantly following intracarotid injection of capsaicin. Small numbers of NO-containing neurons were found in LC, but there was no change in the number of NO-containing neurons in RN and PAG. No NADPH-d histochemical activity could be found in AP. (3) The above responses to capsaicin were significantly inhibited by pretreatment with either a capsaicin receptor antagonist ruthenium red or a NMDA receptor antagonist MK-801. The above results indicate that intracarotid injection of capsaicin may activate the neurons in brainstem nuclei involved in cardiovascular regulation, and that NO only plays an indirect role in the modulation of the responses of brainstem nuclei to capsaicin. These effects of capsaicin are mediated by capsaicin receptors with involvement of glutamate.

There were many factors that affect the chimerism of donor DNA, such as the type of disease, the treatment of the disease, the severity of the disease, the source of hematopoietic stem cells, HLA, GVHD, and so on. This paper mainly discussed the two factors that affect forensic science, including the posttransplantation intervals and sensitivity of the detection method. Forensic workers in forensic practice and paternity and individual identification of allo HSCT recipients, should pay attention to the time after transplantation and the detection method of sensitivity of chimerism. When female recipients received HSCT in the case of male donors, Y chromosome DNA of the donor can be detected from the recipient's hair follicles. The case of biological specimens from the crime scene can not only detect Y chromosome speciifc genetic markers, so as to avoid wrong identiifcation.

Objective To evaluate the clinical value of adenosine triphosphate (ATP) determination in CD4+ cells in cytomegalovirus pneumonia after renal transplantation.Methods The ATP level of CD4+ T cells was measured by ImmuKnowTM kit.The ATP levels were determined in 187 renal transplant recipients before and 30,60,90,180 days after operation,and at the time of CMV pneumonia and 4 weeks after treatment of CMV pneumonia.The associations between ATP levels and CMV pneumonia were analyzed.Analysis of variance (ANOVA),Pearson-Spearman and relative risks were used for data analysis.Results 17 cases out of 187 renal transplant recipients were diagnosed as CMV pneumonia (9.1％),and the onset of CMV pneumonia started on the (2.8 ±1.2)month after renal transplantation.ATP concentrations in CD4+ T cells were significantly lower after operation than those before operation (P＜0.01).ATP concentrations reached the lowest on the about postoperative day 90 (P＜0.05),then increased gradually.In 17 recipients with CMV pneumonia,the ATP levels before and 30,90 days after operation,at the time of CMV pneumonia and 4th week after treatment of CMV pneumonia were (376 ±182),(283 ± 146),(196 ± 112),(145 ± 102) and (236 ± 117) μg/L respectively.ATP levels at the time of CMV pneumonia were significantly lower than any other time points (P＜0.05).There was close correlation between ATP levels and CMV pneumonia.Conclusion The determination of ATP in CD4+ cells could reflect the status of cell-mediated immunity in renal transplant recipients,and could evaluate the severity and prognosis of CMV pneumonia and guide the clinical treatment.

Objective To investigate the usefulness of acoustic radiation force impulse (ARFI) elastography for noninvasive evaluation of liver fibrosis in rats.Methods A total of 70 male Wistar rats were included in the group for dimethylnitrosamine (DMN)-induced liver injury,and 10 saline-injected rats were used as normal control.Hepatic injury was induced by a single intraperitoneal injection of DMN at a dose of 50 mg/kg of body weight.Several rats in the group with DNM injected and the normal control group were randomly selected and sacrificed at each of the following post-injection time:day 5,7,10,14,21,24,and 28.And their livers were taken for pathology analysis.All the rats underwent ARFI elastography before sacrificed in order to acquire a shear wave velocity (Vs) to represent liver stiffness.Correlation between Vs and the histological finding was analysed.ResultsAmong 58 successfully modeled rats,9,13,14 and 12 rats were found to be with S1,S2,S3 and S4 of liver fibrosis pathologically,respectively.And 10 rats were found to be with severe inflammatory activity without any fibrosis.Values of Vs increased with the stage of liver fibrosis ( P ＜0.05).There was a significant correlation between Vs and stage of liver fibrosis ( r =0.947,P =0.000).The areas under ROC curve for the diagnosis of fibrosis S≥S1,S≥S2,S≥S3 and S=S4 were 0.983,0.995,0.999 and 0.964,respectively;for the cutoff values of Vs were 1.59 m/s,2.13 m/s,2.33 m/s and 2.51 m/s,respectively,the sensitivity was 95.8％,92.3％,100％ and 84.6％,and specificity was 100％,100％,96.9％ and 95.6％,respectively.The values of Vs in the group with severe inflammatory activity were significantly higher than those in the control group ( P =0.000).ConclusionsARFI has a relatively high value in the evaluation of liver fibrfosis in rats,while severe inflammatory activity may affect its accuracy.

BACKGROUND: Marrow stromal cells (MSCs) own the characteristic of migration. However, the mechanisms underlying the migration of these cells remain unclear. OBJECTIVE: To explore the roles of stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4 in trafficking of MSCs migration. DESIGN, TIME AND SETTING: The in vivo cytology experiment was performed at Department of Anatomy, National University of Singapore from March 2007 to June 2007. MATERIALS: MSCs were isolated and purified from a Wistar neonatal rat. Forty adult female Wistar rats were randomly divided into sham operation and experimental groups, with 20 animals in each group. METHODS: The chemotaxis assay was performed at a 48 well Boyden chamber, and a total of 25 μL SDF-1 was added to the lower layer of chamber, covered with 8 μm polycarbonate membrane filter; SDF-1 cultured in DMEM conditioned medium was served as a blank control group. Cell concentration was regulated to 1.5×109L-1/L. 50 μL and cell suspension was added into the upper layer of chamber, cultured at CO2 incubator with temperature of 37 ℃ for 10 hours. Rats in the experimental group were prepared for transected spinal cord injury models, and in the sham operation'group, only the vertebral plate was opened. 1.0 mL (1×109L-1/L) MSCs suspension labeled with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) was injected through internal jugular vein at 1 hour after completely transected spinal cord. MAIN OUTCOME MEASURES: Expression of chemokine receptor CXCR4 in MSCs, as well as the effect of SDF-1 on the migration of MSCs was observed by immunofluorescence, change of SDF-1 in lesion site of spinal cord was detected by real-time PCR analysis, as well as the in vivo migration of intravenously injected MSCs was detected by fluorescence microscopy. RESULTS: The pudfied MSCs were positive to CXCR4. Compared to the blank control group, SDF-1 with concentrations of 5, 50, and 500 μg/L could accelerated the migration of MSCs (P < 0.05), which reached a peak with concentration of 500 μg/L. The expression of SDF-1 RNA was obvious increased in the experimental group than that of the sham operation group (P < 0.05), and returned to a normal level at 14 days. At 2 weeks after cell injection, the number of MSCs migrated to the lesion site of completely transected spinal cord was significant increased than sham operation group (P < 0.05). CONCLUSION: SDF-1 may contribute to MSCs migration in vitro and in vivo. SDF-1 and its receptor CXCR4 are involved in the migration of injected MSCs to the lesion site of completely transected spinal cord.Ding P, Xue LP, Yang ZY, Wang CQ, Wang JH, Feng ZT, Ling EA.Chemokine stromal cell-derived factor-1 and its receptor CXCR4 mediate migration of marrow stromal ceils into the lesion site of completely transected spinal cord.