Objective To understand the Helicobacter pylori ( Hp) infection eradication rate of standard tri-ple therapy in Guangdong Meizhou and the drug resistance situation for metronidazole ,clarithromycin ,amoxicillin and levofloxacin ,in order to look for the treatment countermeasures in Hp eradication failure .Methods 297 cases of Hp positive patients because of gastrointestinal symptoms to our hospital examined from April 2011 and March 2013,were randomly assigned into three standard triple therapy groups:A ( OCA ) group and B ( OCM ) group and C ( OCL ) group.The Hp eradication rate was analyzed .Patients with primary treatment failure were selected as group D (OBAL),proceed to (PPl+B+A+L)7 d therapy,the Hp eradication rate was analyzed .230 Hp strains were isola-ted and cultured from 297 cases received the first eradication therapy and 87 cases received again eradication therapy . The minimum inhibitory concentration (MIC) of metronidazole,clarithromycin,amoxicillin and levofloxacin were tested by E-test,in order to determine the resistance of these four antibiotics in clinical isolated Hp strains .Results With intention-to-treat(ITT) analysis,the Hp eradication rates of group A (OCA),group B(OCM) and group C(OCL) were 72.0%(72/100),63.0%(63/100) and 72.2%(70/97),respectively.With per-protocol(PP) analysis,the Hp eradication rates of group A (OCA),group B(OCM) and group C(OCL) were 72.7%(72/99),64.3%(63/98),73.7%(70/95),respectively.The eradication rate among three standard triple therapy groups had no obvi-ous difference (ITT:P=0.278,PP:P=0.288,P>0.05).With ITT analysis,the Hp eradication rate in the quadrup-le therapy group D(OBAL) was 92.0%(80/87).With per-protocol(PP) analysis,the Hp eradication rate in the quadruple therapy group D(OBAL) was 97.6%(80/82),which was higher than that of the three standard triple ther-apy groups(ITT:P=0.000,PP:P=0.000).In 230 clinical isolated Hp strains,the resistant rates of levofloxacin,amoxicillin,clarithromycin and metronidazole were 6.08%(14/230),6.52%(15/230),25.65%(59/230), 70.87%(163/230),respectively.Of those 37 strains were mixed resistance,the mixed resistant rate was 16.09%(37/230).The resistant rate of metronidazole was higher than levofloxacin , amoxicillin and clarithromycin ( P =0.000,P<0.01),the resistant rate of clarithromycin was higher than levofloxacin and amoxicillin (P=0.000),no statistically significant difference between amoxicillin and levofloxacin (P=0.848).Conclusion The Hp resistance is similar to the national average in Guangdong Meizhou ,the eradication rate of standard triple therapy is lower than 80%,contain bismuth agent of quadruple therapy is good rescue therapy .

Objective To study the expression and clinical correlation of miR-141 in peripheral blood and tumor tissue in elderly patients with rectal cancer.Methods Rectal tumor tissue and tumor-adjacent normal tissues were taken in 75 cases of rectal cancer.The expression levels of miR-141 in peripheral blood and tumor tissue as well as tumor-adjacent normal tissues were determined by real-time polymerase chain reaction method.The correlation of miR-141 expression between peripheral blood and tumor tissue,and the correlation of peripheral blood miR-141 with clinicopathologic features and clinical prognosis were analyzed.Results (1) Compared with peripheral blood in the subjects undergoing normal physical examination or tumor-adjacent normal tissue in the patients with rectal cancer,the miR-141 expression level in peripheral blood and tumor tissue in the patients with rectal cancer was decreased significantly (P<0.01).(2) The cancer tissue miR-141 level in the patients with lymph node metastasis was positively correlated with peripheral blood miR-141 level (r=0.694,P<0.01),and cancer tissue miR-141 level in the patients without lymph node metastasis was positively correlated with peripheral blood miR-141 level (r=0.725,P<0.01).(3) The differences between peripheral blood miR-141 level with tumor stage,tumor differentiation degree and lymphatic metastasis had statistical significance (P<0.01).(4)The postoperative 6-month follow up displayed that among 75 cases,13 cases(17.33%) appeared replase/ metastasis,and peripheral blood miR-141 level was (2.64±0.34),which was significantly lower than that in the patents without replase/metastasis (P<0.01).Conclusion Expression variation trend of miR-141 in peripheral blood is similar to that in tumor tissue,which can reflect the clinicopathologic feature in the patients with rectal patients.

Objective To evaluate the eradication rate and long-term therapeutic effect of a triple therapy consisted of cla-(rithromycin) (CLA), amoxicillin (AMO)and omeperazole on Hp infection,and explore the alternative therapeutic programs and their effects after first therapeutic failure.Methods A total of 92 children with Hp infection were divided into two groups: 70 children were given the triple therapy for one week (CLA group);Twenty-two children were given another triple therapy composed of metronida-(zoole) (MET), AMO and omeperazole for two weeks (MET group).All of the children were followed up for 1-30 months after the therapies ended.Children of the two groups who were therapeutic failure were given retreatment as follows.CLA triple therapy were given for one week to the children who were failure after MET triple therapy;increased doses of CLA with longer treatment course was given to the children who were failure after CLA triple therapy . A tetra therapy consisted of colloidal bismuth subcitrate (CBS), furazolidone (FUR) ,omeperazole and AMO was given to children in whom the retreatment failed.Results The Hp eradication rate of CLA group was 91.4%(64/70),and the Hp eradication rate of MET group was 72.7%(16/22).There was significant difference between eradication rate of the two groups(?~2=5.16 P

Catdiomyopathies are diseases that primarily affect the myocardium,leading to serious cardiac dysfimction and heart failure.Out of the three major categories of candiomyopathies(hypertrophic,dilated and restrictive),restrictive cardiomyopathy(RCM)is less common and also the least studied However,the prognosis for RCM is poor as some patients dying in their childhood The molecular mechanisms behind the disease development and progression are not very clear and the treatment of RCM is very difficult and often ineffective.In this article,we reviewed the recent progress in RCM research from the clinical studies and the translational studies done on diseased transgenic animal models.This will help for a better understanding of tare mechanisms underlying the etiology and development of RCM and for the design of better treatments for the disease.

Objective To investigate the clinical characteristics of the human Adenovirus (HAdv) infections in allogeneic hematopoietic stem cell transplantation ( allo-HSCT) patients and explore the clinical significance of HAdv monitoring .Methods A total of 845 cases underwent allo-HSCT were included retrospectively in Perking University People′s Hospital from October 2012 to August 2014.Peripheral blood HAdv load were monitored twice weekly within 100 days after allo-HSCT, or whenever necessary quantitatively by real-time PCR. Meanwhile, other clinical samples such as stool , urine, and bronchoalveolar lavage fluid ( BLAF ) were also detected qualitatively whenever necessary .The follow-up period was at least six months after allo-HSCT.All clinical data were collected and analyzed .Results The total positive rate of HAdv was 3.4% ( 29/845 ) .The incidence of HAdv infection was higher in children [3.8%(6/155), <18y] than that of adults [3.3%(23/690),≥18y].HAdv infection diagnosed within 100 days after allo-HSCT accounted for 72.4%(21/29) of the total number of positive cases .There were 19 cases detected positive in peripheral blood , 16 cases in stool , 9 cases in urine , and 1 cases in BLAF , respectively.One patient was positive in peripheral blood , stool and urine.The overall median time of HAdv was 69 (13-189) d.The median time was 56 (53 -144) d in stool ,which was earlier than that of in peripheral blood , urine and stool.Among 29 cases of HAdv positive patients , 17 patients were coinfected with Cytomegalovirus(CMV) and 11 casess with Epstein-Barr virus(EBV).Twenty-five cases of HAdv were diagnosed with acute graft-versus-host disease(aGVHD) before HAdv infection, and 4 cases were diagnosed with chronic graft-versus-host disease ( cGVHD ) . The most common clinical manifestation was HAdv enteritis (14 cases), followed by hemorrhagic cystitis (7 cases).Two cases complicated with multiple organ injury ( >2 ) clinically, 1 cases with pneumonia.There were 8 cases of death at the end of follow-up.Conclusions HAdv is an important pathogen causing infection in patients after allo-HSCT. The infenction is characterized with multiple organ involvement .CMV and EBV coinfection is common .HAdv monitoring was of great significance in allo-HSCT patients.

A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process.The current investigation is aimed at evaluating the effect of acetyl-L-carnitine on human spermatozoa and reactive species oxygen (ROS) level after freezing-thawing process.The spermatozoa were collected from 35 male patients diagnosed as having asthenospermia.The cryopreservation of human spermatozoa treated with acetyl-L-camitine at different concentrations (group B:2.5 mmol/L,group C:7.5 mmol/L,group D:15 mmol/L) was compared with control (group A:no acetyl-L-carnitine given).For the frozen-thawed spermatozoa,the viability,motility and DNA integrity were measured by comet assay,acrosome integrity by FITC-PNA staining and ROS level was determined in each group.The results showed that there were no significant differences in motility and viability between group A and group B,while the motility and viability of spermatozoa in group C and group D were significantly increased as compared with those in group A.As compared with group A,the values for DNA integrity parameters including comet rate (CR),tail DNA percentage (TD),tail length (TL) and Oliver tail moment (OTM) were significantly reduced in group C and group D.Group C and group D also displayed a higher proportion of intact acrosome than group A.No significant difference in ROS level was found between group A and group B,while with the increase in acetyl-L-camitine concentration,the ROS level in groups C and D was significantly reduced as compared with that in group A.In conclusion,acetyl-L-camitine at a concentration of 7.5 mmol/L is an effective antioxidant against cryo-damage on post-thawed human spermatozoa.

It has gradually become a consensus in the industry that the traditional Chinese medicine gypsum should be decocted first, but the understanding of decocting method is not completely unified in the works of doctors since ancient times, and there are occasional disputes about whether it is necessary to decocting first. In this study, the phase determination, physical and chemical characterization, qualitative and quantitative analysis of inorganic and organic components of the decoctions of herbal pairs and the whole prescription Maxingshigan decoction with gypsum as the center, and the pre-decoctions and co-decoctions of them were carried out to explore the scientific connotation of the pre-decoctions of gypsum. Results show that decoction phases were different between the co-decoctions and pre-decoctions of licorice-gypsum (Gancao-Shigao, GC-SG), ephedra-gypsum (Mahuang-Shigao, MH-SG) and almond-gypsum (Xingren-Shigao, XR-SG). The results of the micromorphology, particle size and zeta potential of herbal pairs and prescription (Quanfang, QF) showed that the supramolecular particles in pre-decoctions were smaller, more uniform and more stable than the co-decoctions. The results of organic components analysis showed that different cooking methods did not change the organic composition and content. ICP-OES results showed that the content of inorganic components in pre-decoctions was higher than in co-decoctions for the same boiling time of gypsum. The IR results showed that the pre-decoctions had stronger chemical functional group effect than the co-decoctions. To sum up, compared with the co-decoction, the pre-decoction of gypsum has different phase state and chemical composition interaction, and the difference of inorganic composition is an important material basis affecting the change of phase state compared with the co-decoction. It indicates that the material basis of traditional Chinese medicine decoction is indeed different whether gypsum is decocted first or not, which can provide a basis for the clinical application of decocted gypsum.

Objects To explore the association of ulcerative colitis (UC) with the imbalance between Th1,Th 2 and Th17 cells in the colonic tissues.Methods A total of 41 UC patients and 52 controls was recruited in the present study.The real-time fluorescent quantitative PCR was applied for detecting the mRNA levels of Thl,Th2 and Th 17 cells-associated transcription factors T-bet,GATA-3 and RORγt and cytokines IFN-γ,IL-4 and IL-17A in the colonic tissues.Simultaneously,the expressions of IFN-γ,IL-4 and IL-17A in the colonic tissues were also examined by an immunohistochemical staining method.Results Compared with the controls,the mRNA expressions of GATA-3,RORγt and IL-17A were more significantly enhanced in UC patients (0.84 ± 0.24 vs.0.69 ± 0.22,P=0.002;0.99 ± 0.29 vs.0.83 ± 0.23,P=0.004;1.59 ± 0.65 vs.1.35 ± 0.43,P=0.035).According to the ＂Truelove and Witts Severity Index＂,those patients were divided into different subgroups.The mRNA expressions of GATA-3,RORγt,and IL-17A were shown to be higher in patients with moderate and severe UC than in those with mild UC (0.90 ± 0.18 vs.0.78 ± 0.16,P=0.030;1.11 ± 0.31 vs.0.87 ± 0.26,P=0.011;1.83 ± 0.64 vs.1.34 ± 0.66,P=0.020).Moreover,the immunohistochemistry results demonstrated that the IL-17A positive cells were positioned mainly in the intestinal epithelial layer and lamina propria.Compared to the controls,the mean integral optic density of IL-17A was significantly increased in the colonic tissues of UC patients (0.25 ± 0.07 vs.0.13 ± 0.03,P＜0.001).The similar results were obtained for IL-17A in patients with moderate and severe UC when compared to those with mild UC (0.31 ± 0.07 vs.0.19 ± 0.06,P＜0.001).In contrast to the controls,the mRNA ratios ofGATA-3/T-bet,RORγt/ T-bet and RORγt/GATA-3 were significantly higher in the tissues of colonic UC patients (1.12 ± 0.30 vs.0.96 ± 0.31,P=0.014;1.33 ± 0.37 vs.1.15 ± 0.33,P=0.015;1.44 ± 0.45 vs.1.20 ± 0.42,P=0.009),and in the patients,the mRNA ratios for GATA-3/T-bet,RORγt/T-bet and RORγt/GATA-3 were significantly higher in the patients with moderate and severe UC than in those with mild UC (1.27 ± 0.35 vs.1.00 ± 0.32,P＜0.001;1.45 ± 0.37 vs.1.19 ± 0.36,P=0.028;1.59 ± 0.43 vs.1.28 ± 0.46,P=0.031).Conclusions These findings suggest that the imbalance between Thl,Th2 and Th17 cells in the colonic tissues may be implicated in UC.

OBJECTIVEIn order to accumulate experiences for improving the efficiency in serological tests, the present study on mixed testing of serum samples was performed by taking the serological test of trichinellosis and toxoplasmosis as the examples, and had proved the effects on cost-effectiveness of seroepidemiological survey of parasitic disease with method of mixed-samples test.

METHODSAccording to the binomial distribution principle, to develop an approach to the feasibility of mixed testing of serum samples, and to work on a cost-effectiveness analysis of one-by-one testing and mixed testing using hygienic economic analysis method was performed. For serological test of trichinellosis and toxoplasmosis, 3 kinds of mixed testing methods, namely 3 serum sample mixture, 5 serum sample mixture and 10 serum sample mixture, were performed.

RESULTSThe results showed that all the 3 kinds of mixed tests of trichinellosis and toxoplasmosis showing positive result if only 1 weak positive serum sample were mixed with. When the serum samples being mixed were all negative ones, then among the 24 groups tested with each kind of negative serum sample mixture of trichinellosis (3 serum samples, 5 serum samples and 10 serum samples), they all showed negative. However, among the 12 groups tested with 2 kinds of negative serum mixture of toxoplasmosis (3 serum samples and 5 serum samples), all showed negative while among the 18 groups tested with the 10 serum sample mixture, 16 groups showed negative and 2 were positive. The mixed testing of trichinellosis and toxoplasmosis showed that the efficiency of mixed testing was related to the serological positive rate of the parasitic diseases to be examined. When serological positive rate was 10%, the efficiency of mixed testing was higher in 4 serum sample group. When serological positive rate was 1%, the efficiency of mixed testing was higher in 10 serum sample group and when serological positive rate was 0.1%, the in crease of the size of mixed serum samples could decrease the number of testing, but the prerequisite was that there must be one positive sample, so that the positivity for all the mixed tests could be detected. If mixed testing were performed on all negative samples, no positivity could be detected.

CONCLUSIONThe result of cost-effectiveness analysis demonstrated that for seroepidemiological survey of parasitic diseases, the cost for mixed testing was low, especially when the serological positive rate was expected low (< or = 1%, thus the mixed testing could save a large amount of the cost.

Cost-Benefit Analysis ; Data Collection ; Humans ; Seroepidemiologic Studies ; Specimen Handling ; Toxoplasmosis ; diagnosis ; epidemiology ; Trichinellosis ; diagnosis ; epidemiology

Classical swine fever virus (CSFV), an enveloped positive-stranded RNA virus in the genus Pestivirus of the Flaviviridae family, is the causative agent of a highly contagious swine disease characterized by symptoms of hemorrhagic fever and immune depression, usually leading to substantial economic losses. The serological methods for detection of CSFV antibody such as ELISA are important means for the diagnosis of CSFV and immune surveillance. It is difficult to obtain CSFV antigen with high quality using traditional method because its titration titer is low in cell culture. CSFV has four structural protein named C, E0, El and E2. The E2 protein contains major antigenic determinants that are conserved between different CSFV strains and involved in neutralization by antibodies. So recombinant E2 protein can be developed as an alternative to the intact viral antigen. So far, CSFV E2 have not been expressed in E. coli with high level. Many factors, such as the secondary structure, the stability of 5' and 3' terminus of gene, the location of SD sequence and the bias of codes, are involved in the expressing level of foreign gene in E. coli . In this study, two sites of the E2 gene sequence were confirmed to be detrimental to its expression efficiency in E. coli through the computer-aided analysis. So they were mutated using recombinant PCR without changing the amino acids sequence of CSFV E2 gene. A plasmid was constructed by inserting the mutated E2 gene into the prokaryotic expression vector pET-28a(+) and named pETE2. The E. coli competent host BL21 (DE3)lysS transformed with pETE2 could express the E2 gene at high level, amounting to 28% of the total protein of the induced recombinant bacteria at the presence of IPTG. Except the hydrophobic transmembrane domain at C terminus, the recombinant E2 protein includes the total aa sequence. So it contains all the potential linear antigen epitopes of E2 protein because hydrophobic aa region can not form epitope. The recombinant E2 protein was CSFV-specific as proved by Western blotting and indirect ELISA. The rabbits immunized with the recombinant E2 can be protect from the challenge of hog cholera lapinized virus. This is the first report that E2 gene is expressed with high level expression in E. coli. In conclusion, it is an effective measure that mutate the CSFV E2 gene to increase its expression level in E. coli. The recombinant CSFV E2 protein possess fine immunonicity and can be used the antigen for the detection of CSFV antibody.
Antigens, Viral ; genetics ; immunology ; metabolism ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Mutagenesis, Site-Directed ; methods ; Polymerase Chain Reaction ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Viral Envelope Proteins ; genetics ; immunology ; metabolism