Background Recombinant hirudin variant Ⅲ(rHV3) can effectively prevent galactose-induced human lens epithelial cells LECs injury,but little is known about the molecular mechanism of its action.Objective The present study was to investigate the effects of rHV3 on the expression of apoptosis-related genes in damaged LECs induced by galactose.Methods The rHV3 was extracted by our research group,and the biological activity of rHV3 was identified by titration of thrombase according to Markwardt's method.Human LECs (SRA01/04) were cultured using 125×10-3 mol/L D-galactose+10% FBS+D/F12 medium to establish the damaged human LECs model.rHV3 was added into the medium of the damaged human LECs model.Human LECs were cultured in D/F12 medium containing 10% FBS as normal control.The expression of apoptosis-related genes,such as aldose reductase (AR),bax,bcl2 and p53,in LECs at the mRNA level was detected using RT-PCR.The abundance ratio of target genes was presented with the absorbance (A) of gene mRNA/GAPDH mRNA.Results Compared to the normal control group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were significantly elevated in model group (t=3.90E-06,t=8.44E-04,t=5.15E-08,P＜0.01).However,in the rHV3-treated group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were lower than those of model group (t=5.90E-06,t=1.51E-04,t=3.42E-06,P＜0.01).The bcl2 mRNA/GAPDH mRNA was markedly downregulated in the model group when compared with the normal control group (t=1.86E-05,P＜0.01);while after rHV3 addition,bcl2 mRNA/GAPDH mRNA increased in comparison with the model group (t=8.56E-05,P＜0.01).Conclusion 125×10-3mol/L D-galactose induces the damage and apoptosis of human LECs.rHV3 likely plays a protective function on D-galactose-induced damage of human LECs by inhibiting the polyol pathway and mitochondria-mediated pathway.

OBJECTIVETo evaluate the clinical efficacy and safety of acupuncture-moxibustion in treatment of irritable bowel syndrome systematically.

METHODSClinical randomized controlled trials on treatment of irritable bowel syndrome with acupuncture-moxibustion were collected. Through retrieval of CNKI (1979 - December of 2011) and VIP (1979- December of 2011), randomized and quasi-randomized controlled clinical trials on treatment of irritable bowel syndrome with control study between acupuncture and sham acupuncture or western medication were included. The test bias risk and quality assessment of each experiment were carried out by two researchers in accordance with the Cochrane Handbook 5.1.0 standard. And RevMan 5.1.6 software was adopted for the Meta analysis.

RESULTSEleven researches were included with totally 969 patients. Meta analysis shows that the effective rate of the combined methods of acupuncture and moxibustion [RR = 1. 27, 95% CI ( 1.09, 1.49)] is superior to conventional western medication treatment.

CONCLUSIONAcupuncture-moxibustion for irritable bowel syndrome is better than the conventional western medication treatment.

Acupuncture Therapy ; Clinical Trials as Topic ; Humans ; Irritable Bowel Syndrome ; therapy ; Moxibustion

In order to study the value of sequential and quantitative analysis of chimerism in determination of optional time of donor lymphocyte infusion (DLI) and prediction of efficacy of DLI, six patients with leukemias who relapsed or failed of engraftment were treated with DLI. Serial and quantitative analyses of donor chimerism (DC) both prior to and following DLI were performed by multiplex PCR amplification of STR markers (STR-PCR) and capillary electrophoresis with fluorescence detection. The results showed that at the time of relapse or graft rejection, STR-PCR indicated the decreasing donor chimerism in all six patients, at levels ranging from 27.3% to 85.7%. The declining value of DC (<90%) was detected in four patients at 26 days before relapse or graft rejection diagnosed clinically. Therefore the decrease of value of DC can be identified the high risk of relapse or graft failure and can be used to guide DLI implementation at early stage. In this study the clinical response were seen in two patients, the value of DC in these patients increased with convertion to a predominant donor profile (>90%) or converted to stable FDC shortly after DLI, while in the patients without clinical response, the level of DC decreased persistently or declined after transient increase. Three patients without response received second DLI. It is concluded that the monitoring of chimerism is proved to be a valuable to determine the optional time point of DLI and to early evaluate the efficacy of DLI. Furthermore, it can present a rational basis for treatment of intensification in the patients who did not respond to first-line DLI treatment.
Adolescent ; Adult ; Hematopoietic Stem Cell Transplantation ; Humans ; Lymphocyte Transfusion ; Recurrence ; Tissue Donors ; Transplantation Chimera

In order to evaluate relapse predication ability of STR-PCR combining with qualitative RT-PCR for the bar/abl transcripts to the patient with chronic myeloid leukemia (CML) fulfilled allogeneic stem cell transplantation (allo-HSCT), 24 patients with CML after allo-HSCT were dynamically investigated for MRD, quantitative analysis of donor chimerism was performed by multiplex PCR amplification of STR markers and capillary electrophoresis with fluorescence detection, qualitative detection of bcr/abl transcripts was detected by nested RT-PCR. The results showed that persistent full donor chimerism (DC >/= 95%) was associated with an absence of MRD. All patients with stable MC (90% Genes, abl ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; diagnosis ; therapy ; Neoplasm, Residual ; Polymerase Chain Reaction ; methods ; Transplantation, Homologous

OBJECTIVETo evaluate the early operative treatment and clinical results of pelvic fractures associated with urethra disruption.

METHODSFrom January 1995 to January 2005, 25 patients suffered from pelvic fractures combined urethra disruption treated by operation were retrospectively analyzed. According to Tile's classification, 1 case was stable pelvic fracture, 17 rotational unstable fractures, and 7 rotational combined vertical unstable fractures. The complete urethra rupture were in 23 cases and incomplete in 2 cases. The operative methods included: (1) emergency open reduction and internal fixation of the pelvis combined primary urethra suturing in 2 cases, partial suturing after realignment in 4 cases, realignment in 2 cases, and urethrovaginal penetrating wound repairing in 1 case; (2) primary urethra realignment only and delayed (range, 7 to 21 days) pelvic internal fixation in 10 cases; (3) early cystostomy and delayed (range, 3 to 21 days) urethra realignment and pelvic internal fixation in 6 cases.

RESULTSThe mean follow-up time of all patients was 34 months (range from 6 to 120 months). According to Majeed's evaluation, 17 cases of pelvic injury showed excellent results, 5 good, and 3 fare. After urinary catheter removed, the mean maximal urine flow rate of 19 (76%) patients was 18.6 ml/s and the mean scar length between both disrupted ends on the film of excretion urethrography was 0.51 cm. Five (20%) cases suffered in dysuria needed urethral dilatation or further surgery. One (4%) female could not control urination who need a second-look operation. The primary suprapubic soft tissue avulsion wound infection secondary to retropubic abscess was found in 1 case, posterior urethra-stenosis in 5 cases, sexual impotence in 3 cases, and incontinence in 1 case.

CONCLUSIONSThe satisfactory reduction and effective fixation of the pelvic fractures is an anatomical basis for receiving "tension-free urethral anastomosis".

Adult ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Pelvic Bones ; injuries ; Postoperative Complications ; prevention & control ; Retrospective Studies ; Time Factors ; Treatment Outcome ; Urethra ; injuries

Purpose :To investigate the relationship of peripheral blood T-lymphocyte subsets with the acuterejection or severe CMV infection in transplanted patients. Methods :T-lymphocytes subsets of peripheral bloodwere consecutively detected by using mice-verse-human T-lymphocytes subsets monoclonal antibody-OKT serialsand flow cytometer. Results:The difference of CD4/CD8 ratios between the no acute rejection group and the acuterejection group, or between the acute rejection remission group and the resistant acute rejection group was signif-icant ( P ＜0.05); In patients with intensive CMV infection, the CD4/CD8 ratios were converse to the acute re-jection group. Conclusions:These results indicated that monitoring of peripheral blood T-lymphocyte subsets wasof much benefit to early diagnosis and differential diagnosis of acute rejection of intensive CMV infection and rea-sonable treatment.

ABSTRACTOBJECTIVE To explore the acupuncture treatment mechanism of constipation?predominant irritable bowel syn-dromeIBS?C from the perspective of brain?gut axisby observing the plasma 5HTCGRP and NPY level before and after the treatment.METHODS 60 patients with IBS?C were randomly divided into two groups.30 cases in the acupuncture groupAG were treated by acupuncture five times a weekwith such acupoints as TianshuZusanliShangjuxuTaichongSanyin-jiaoYintangBaihui being selected.The other 30 cases in the medicine groupMGwere treated with lactulose oral solution15 ml per time for three times a day.After 4 weeks treatmentenzyme?linked immunosorbent assayELISAwas used to de-tect the plasma 5HTNPY and CGRP level of 30 healthy volunteers and the two treatment groups and observe the improve-ment of clinical symptoms.RESULTS ①Both groups could significantly improve the clinical symptoms in patients with IBS?CP <0.01while the curative effect in the AG was superior to that in the MGP <0.01.②The levels of 5?HTNPY and CGRP in patients with IBS?C of two treatment groups were significantly higher than those in healthy volunteersand the differ-ence was statistically significantP <0.01.The plasma 5?HTNPY and CGRP levels all went down in both groups after treatmentand the differences of 5?HTNPY and CGRP levels were significant when compared to the levels before treatment in AGP <0.01.The differences of 5?HTNPY levels were also significant when compared to the levels before treatment in MGP <0.01while there was no significant difference on CGRPP <0.01.There were no significant differences of 5?HT and NPY levels between these two groups after treatmentP >0.05but the difference of CGRP was highly statistically sig-nificantP <0.01.The downward trend of 5?HTNPY and CGRP level was better in the AG.CONCLUSION ①Acupunc-ture treatment can significantly improve the clinical symptoms in patients with IBS?Cand the theraputic effect is superior to that in the MG.②The increase of 5?HTNPY and CGRP level in patients with IBS?C after treatment suggests a close relation-ship between the abnormal level of brain?gut peptide and IBS syndrome.③Acupuncture treatment can decrease the plasma lev-els of 5?HTNPY and CGRPalleviate the degree of abdominal pain and discomfortwhich indicates that benign regulation of brain?gut peptide level in patients with IBS?C by acupuncture may be one of the effective mechanisms in treatmenting this dis-ease.

BACKGROUNDInflammation plays a pivotal role in the formation and progression of ischemic stroke. Recently, more and more epidemiological studies have focused on the association between C-reactive protein (CRP) -717A > G and -286C > T > A genetic polymorphisms and ischemic stroke. However, the findings of these researches are not conclusive.

METHODSWe performed a meta-analysis to determine whether these two polymorphisms are associated with the risk of ischemic stroke. Eligible studies were identified from the database of PubMed, Medline, Embase, Web of Science, CNKI, Weipu, and Wanfang. We calculated odds ratios (ORs) with 95% confidence intervals (CIs) to assess the strength of the association.

RESULTSFour articles were included in our study, including 1926 cases and 2678 controls for -717A > G polymorphism, 652 cases and 1103 controls for -286C > T > A polymorphism. The results of meta-analysis showed that single nucleotide polymorphism (SNP) -717A > G was not significantly associated with the risk of ischemic stroke (GG vs. AA, OR = 1.12, 95% CI = 0.83-1.50, P = 0.207; GG + GA vs. AA, OR = 1.04, 95% CI = 0.93-1.17, P = 0.533; GG vs. GA + AA, OR = 1.10, 95% CI = 0.82-1.47, P = 0.220). Meta-analysis of SNP - 286C > T > A also demonstrated no statistical evidence of a significant association with the risk of ischemic stroke (AA vs. CC, OR = 0.86, 95% CI = 0.59-1.25, P = 0.348; AA vs. CC, OR = 0.92, 95% CI = 0.80-1.06, P = 0.609; AA vs. CC, OR = 0.89, 95% CI = 0.62-1.30, P = 0.374).

CONCLUSIONSThis meta-analysis demonstrated little evidence to support a role of CRP gene -717A > G, -286C > T > A polymorphisms in ischemic stroke predisposition. However, to draw comprehensive and more reliable conclusions, further larger studies are needed to validate the association between CRP gene polymorphisms and ischemic stroke in various ethnic groups.

Alleles ; Brain Ischemia ; genetics ; C-Reactive Protein ; genetics ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Polymorphism, Single Nucleotide ; genetics ; Stroke ; epidemiology ; genetics

The objective of this study was to investigate the alterations of cytokines TNF-alpha, IL-1beta and IFN-gamma at early phase after allogeneic hematopoietic stem cell transplantation and in the course of preconditioning, and to explore the relation of these cytokines with transplant-related complications. Alterations of TNF-alpha, IL-1beta, and IFN-gamma levels in serum were detected by ELISA in 95 patients received allogeneic hematopoietic stem cell transplantation (among them 43 cases with GVHD, 5 cases with thrombosis, 31 cases with infection) and 20 in healthy adults. Alterations of the three cytokines were analyzed during the preconditioning and the early phase after transplantation. The results showed that the TNF-alpha levels in aGVHD patients underwent allo-HSCT were already higher than that in normal controls before preconditioning (p<0.01), other patients did not show significant change during this course. TNF-alpha level in all patients were higher than that at day 4 of preconditioning, then decreased at end of preconditioning (p<0.05). TNF-alpha level increased at occurrence of aGVHD, thrombosis and infection, which is most significant in patients with aGVHD, and less significant in patients with infection as compared with patients with thrombosis (p<0.05). TNF-alpha level began to increase at 2 weeks before aGVHD and thrombosis developed in patients, while TNF-alpha levels did not change in patients with infection at the same time. IL-1beta levels did not change during preconditioning, but increased at time of aGVHD, thrombosis and infection in patients, in which IL-1beta levels in patients with thrombosis increased obviously, and more obviously in patients with aGVHD than that in patients with infection (p<0.01). IL-1beta levels in patients with aGVHD began to increase at 1 week before aGHVD developed, but IL-1beta levels in patients with thrombosis began to increase at two weeks before complication developed. IFN-gamma levels did not change in all patients during the process of transplantation. It is concluded that the alterations of cytokine levels exist during the course of allo-HSCT, which reflects the vascular damage following preconditioning and occurrence of some transplant associated complications. Levels of TNF-alpha and IL-1beta are closely related to aGVHD or thrombotic complications. Monitoring changes of TNF-alpha and IL-1beta levels contributes to early discovery of aGVHD and thrombotic complications.
Adolescent ; Adult ; Child ; Female ; Graft vs Host Disease ; diagnosis ; Hematopoietic Stem Cell Transplantation ; Humans ; Interferon-gamma ; metabolism ; Interleukin-1beta ; metabolism ; Male ; Middle Aged ; Thrombosis ; diagnosis ; Transplantation Conditioning ; methods ; Transplantation, Homologous ; Tumor Necrosis Factor-alpha ; metabolism ; Young Adult

OBJECTIVETo establish multiple short tandem repeat (STR) amplification by fluorescence labeling polymerase chain reaction (PCR) combined with capillary electrophoresis for quantitative determination of chimerism, and to evaluate the status of engraftment and predict the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSThirty-one patients received bone marrow transplantation (BMT) or nonmyeloablative allogeneic stem cell transplantation (NST) were evaluated. Peripheral blood and bone marrow were co-llected before and after transplantation in different period. Nine different STR markers were co-amplified in a single reaction by using a commercial AmpF/STR Profiler Plus PCR amplification kit. Separation of the PCR products and fluorescence detection were performed by ABI prism 310 Genetic Analyzer with capillary electrophoresis. The Genescan and Genotype software were used for size calling and quantification of peak areas. The formula to calculate donor chimerism values was based on the different allelic distribution type between donor and recipient.

RESULTS48.4% of the patients received sex-matched transplantation and the quantification of donor chimerism could only be performed by STR-PCR method. Comparison of values obtained by FISH analysis with that by STR-PCR in patients transplanted from sex-mismatched donors showed an excellent correlation. The median number of informative alleles was 6.7 (range 2 - 10). The donor's alleles appeared in all the patients on day 7 post-transplant. The median values of donor chimerism in BMT group were inferior to that in NST group on day 7, day 14 and 1 month post-transplant. However the difference disappeared in the midterm or later period of transplant. On day 21, all of the 31 patients had stable engraftment and the percentage of donor chimerism was more than 92%. Median follow-up was 17 (3.5 - 29.0) months after transplantation. Twenty-six of 31 patients had durable engraftment and donor chimerism ratio was more than 90%. So for all of them survived leukemia-freely. Four of the 31 patients had unstable mixed chimerism and relapsed within 6 months post allo-HSCT. Another patient with unstable mixed chimerism appeared graft rejection. Decreasing values of donor chimerism were detected prior to the occurrence of graft rejection and disease relapse. The incidence of GVHD was much higher in the group of full donor chimerism.

CONCLUSIONSequential and quantitative monitoring of STR is a valuable tool for studying engraftment dynamics, graft rejection, and relapse and for predicting GVHD. Furthermore it can provide a basis for early intervention of clinical treatment.

Adolescent ; Adult ; Child ; Electrophoresis, Capillary ; Female ; Graft Rejection ; Hematopoietic Stem Cell Transplantation ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Polymerase Chain Reaction ; Recurrence ; Tandem Repeat Sequences ; Transplantation Chimera ; Transplantation, Homologous