Microbiology experiment existing independently from microbiology theoretical curriculum is an indispensable compulsory course in contemporary life science. This article presents the principle applied by the National Excellent Microbiology Course teaching group in Nankai University, which is to strengthen the undergraduates’ basic skills of conducting microbiology experiments. With an aim to enhance the core competitiveness of the undergraduates, we have established the three-level experimental contents. A new multilevel teaching pattern focusing on basic skill training as the cornerstone has been applied to enhance the overall competences of the students and to stimulate their innovation abilities. Students’ experimental accomplishment will also be taken into consideration when their experiment results are evaluated, which helps to standardizing their research ethics.

Objective To study the percutaneous permeability through mouse skin of lidocaine hydrochloride-loaded destran-based niosomes(LID-HLD-BNs)in vitro and in vivo. Methods HPLC was employed to exam lidocaine hydrochlo?ride. Lidocaine hydro-chloride-loaded conventional liposomes (LID-CLs) and lidocaine hydrochloride injection (LID-IJ) were used as control. Isolated mouse skin was added into Franz diffusion cell to evaluate the permeability of LID-HLD-BNs in vitro. Confocal Laser Scanning Microscopy(CLSM)was used to observe the permeation depth of mouse skin in vivo. Re?sults The permeation rate and cumulative permeation amount were significantly higher in LID-HLD-BNs group than those of LID-CLs and LID-IJ groups (P<0.05). CLSM studies also confirmed that HLD-BNs reached deeper layers of the skin. Conclusion LID-HLD-BNs has good transdermal ability.

OBJECTIVETo compare the effect of transumbilical single-site single-port with that of transumbilical single-site double-port laparoscopic varicocelectomy in the treatment of varicocele in adolescents.

METHODSWe randomly assigned 80 varicocele patients aged 10 - 16 years to two groups of equal number to receive transumbilical single-site single-port and single-site double-port laparoscopic varicocelectomy, respectively. We compared the operation time, postoperative hospital stay, incisional pain, complications and satisfaction with the abdominal cosmetic outcomes between the two groups.

RESULTSAll the operations were successfully performed. The double-port group showed a significantly higher score on the Visual Analogue Scale than the single-port group (4.8 +/- 1.4 vs 3.6 +/- 1.1, t = -4.986, P < 0.01), but there were no significant differences between the two groups in the operation time ([29.8 +/- 4.2] vs [31.2 +/- 4.6] min, t = 1.383, P = 0.171), postoperative hospital stay ([1.95 +/- 0.7] vs [1.82 +/- 0.8] d, t = -0.784, P = 0.436), complications (0 vs 0) and scores on the satisfaction with abdominal cosmetic outcomes (4.6 +/- 0.6 vs 4.8 +/- 0.5, t = 1.253, P = 0.214). No recurrence, umbilical hernia, hydrocele and orchiatrophy were found in the two groups of patients at 6 months after operation, and no visible scar was observed on the abdominal surface.

CONCLUSIONWith strict surgical indications, single-site single-port and single-site double-port laparoscopic varicocelectomies have similar clinical effects in the treatment of varicocele, which leave no scar on the abdominal surface. Single-site double-port laparoscopy needs no special instruments and therefore is worthier of wide clinical application.

Adolescent ; Child ; Humans ; Laparoscopy ; methods ; Length of Stay ; Male ; Operative Time ; Umbilicus ; surgery ; Varicocele ; surgery

Objective To investigate the effect of arsenic trioxide(As_2 O_3)nanoparticles on rabbit vascular smooth muscle cells in vitro in comparison with normal form As_2 03.Methods The rabbit vascular smooth muscle cells were cultured in vitro.Nano and normal forms of As_2O_3 with drug concentrations of 3?mol/L were added into the cells.Cell proliferation curve was drawn according to the light absorption values of MTT test.Flow cytometry was applied to observe the apoptosis.DNA was extracted and underwent electrophoresis.Results Cell proliferation treated with the 3?mol/L concentration of As_2O_3 was inhibited. Cell growth was inhibited markedly with increased treatment time,and the inhibition effect of nano drug form seemed stronger than that of normal form.MTT light absorption values of cells treated at 24,48 and 72 h showed statistically significant difference(H=10.934,15.039,15.539,P

Objective To understand the perceived stigma and discrimination and received stigma and discrimination as well as the differences and reasons of them among people living with HIV/AIDS and their family members. Methods 307 people were investigated and 117 of which were HIV/AIDS and the others were their family members. Face to face interview was carried out. Results HIV/AIDS and their family were living in the social environment rounded by stigma and discrimination. Stigma and discrimination could be divided into perceived stigma and discrimination and received stigma and discrimination, and these two kinds of stigma and discrimination were statistically significant different between HIV/AIDS and their family members (t=-12.540, P=0.000) , and the perceived stigma and discrimination was more serious than received stigma and discrimination. The perceived stigma and discrimination were related to their self-efficacy (OR=0.558, P=0.041 ), family APGAR (OR= 0.650, P=0.027), infected with HIV or not (OR= 2.116,P=0.004). The received stigma and discrimination were related to their self-efficacy (OR=0.468,P=0.028), family APGAR (OR=0.427, P=0.000) whether infected by HIV (OR=3.412, P=0.001 ). Conclusion Stigma and discrimination did exist in the environment where HIV/AIDS and their family lived, suggesting that it was necessary to develop a series of policies and measures to fight against stigma and discrimination.

OBJECTIVETo develop a real-time quantitative polymerase chain reaction(PCR) assay for detecting Rickettsia rickettsii.

METHODSThe primers and TaqMan-MGB probe were designed according to the ompB gene of R. rickettsii. A DNA fragment of ompB gene amplified from R. rickettsii by PCR was used as a standard template for the development of the method.

RESULTS5 copies of ompB fragments of R. rickettsii were detected. The genomic DNA of R. rickettsii was detected by the developed quantitative PCR assay. However, the genomic DNA from another rickettsial or bacterial agent was not determined. Through this developed method, the positive results were obtained from the animals and cells, artificially infected with R. rickettsii.

CONCLUSIONThe real-time quantitative PCR assay seemed to be highly sensitive and specific which might be used to rapidly detect R. rickettsia DNA in various samples and to early diagnose patients infected by R. rickettsii.

DNA Primers ; Humans ; Polymerase Chain Reaction ; methods ; Rickettsia rickettsii ; genetics ; Rocky Mountain Spotted Fever ; diagnosis ; Sensitivity and Specificity

OBJECTIVETo develop a quantitative real-time polymerase chain reaction (PCR) for detecting Bartonella henselae.

METHODSAccording to the 16S-23S rRNA intervening sequences (IVS) specific for B. henselae, one pair of primers and one TaqMan-MGB probe were designed. A quantitative real-time PCR was developed with the primers, the probe, and the IVS, a standard template, in DNA sequence detection system (ABI 7900HT).

RESULTSThe standard curve was established with the standard template and the relationship between the value of threshold cycle (Ct) and the DNA copy number was linear (r = 0.997). The sensitivity of this quantitative real-time PCR was about 1000 times higher than that of a common PCR used to detect homologous DNA. By this quantitative real-time PCR, the DNA sample of B. henselae was positively detected but not from other rickettsial or bacterial DNA samples. The variation coefficients of intra- and inter-assay reproducibility were 0.2%-1.9%. Using the real-time quantitative PCR to detect samples from mice that were experimentally infected with B. henselae, the small amount of B. henselae DNA was detected in blood samples on days 2, 3, and 5 and large amount of B. henselae DNA was detected in spleen samples on days 1 and 2 after infection.

CONCLUSIONResults from our study suggested that this quantitative real-time PCR was highly specific, sensitive and with good repeatability for detection of B. henselae. It seemed quite useful for rapid detection of tiny DNA of B. henselae in various samples and laboratory diagnosis of bartonellosis caused by B. henselae.

Animals ; Bartonella Infections ; diagnosis ; Bartonella henselae ; genetics ; DNA, Bacterial ; analysis ; Mice ; Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVE: To study the association between clinical phenotypes and genotypes in children with Becker muscular dystrophy (BMD)/Duchenne muscular dystrophy (DMD) so as to provide a theoretical basis for disease management, gene therapy, and prenatal diagnosis. METHODS: A retrospective analysis was performed for the clinical data and gene detection results of 52 children with BMD/DMD. Multiplex ligation-dependent probe amplification (MLPA) was used to detect the DMD gene. The children with negative results of MLPA were further screened by exon chip capture combined with next-generation sequencing (NGS). The mothers of 20 probands were validated by sequencing. RESULTS: The pathogenic genes for BMD/DMD were detected in 50 children by MLPA and NGS, with a detection rate of 96%. Among the 52 children, 36 (69%) had gene deletion, 7 (13%) had duplication, and 7 (13%) had micromutation. Among the 43 children with deletion/duplication, 32 had DMD and 11 had BMD; 37 children (86%) met the reading frame rule, among whom 27 (96%) had DMD and 10 (67%) had BMD. All 7 children with micromutation had DMD. CONCLUSIONS The reading frame rule has an extremely high predictive value for DMD but a limited predictive value for BMD.
Child ; Dystrophin ; Female ; Gene Deletion ; Genotype ; Humans ; Multiplex Polymerase Chain Reaction ; Muscular Dystrophy, Duchenne ; Mutation ; Phenotype ; Pregnancy ; Retrospective Studies

Objective To investigate the prognostic factors affecting the postoperative survival of patients with invasive bladder cancer,and to predict the survival time of the patients.Methods We retro- spectively analyzed the clinical and follow-up data of 178 patients with invasive bladder cancer treated by radical cystectomy and urinary diversion from 1991 to 2004.A multivariate analysis was performed in these patients by the Cox proportional hazard model.A prognostic index(PI)based on the Cox regression was con- structed.According to the individualized PI,the patients were classified into different hazard groups and the expected survival curve of each patient was calculated.Results Cox regression analysis showed that the factors which influenced the postoperative survival included tumor stage(RR=1.982,P=0.000),grade (RR=1.978,P =0.042),lymph node metastasis(RR=2.142,P=0.048),Tis(RR=6.177,P= 0.000),tumor shape(RR=0.416,P=0.003),number of tumors( RR=1.820,P=0.035),pathological type(RR=2.228,P=0.032),patient age(RR=0.672,P=0.025)and neoadjuvant chemotherapy (RR=0.257,P=0.016).Based on the percentile of PI,patients were classified into 3 prognostic groups; the median survival time of 3 groups were 42.5,22.5 and 7.0 months,respectively.There were significant differences between each 2 among the 3 groups(P＜0.01).Conclusions Neoadjuvant chemotherapy, tumor stage,grade,lymph node metastasis,Tis,shape and number of tumors,pathological type,patient age were important prognostic factors.PI value can be used to predict the prognosis of patients with invasive blad- der cancer.

OBJECTIVETo study the relation between the nitric-oxide synthase (NOS) expression and nitric oxide (NO) content in the skeletal muscles and the injury condition of soft tissue in the 3rd lumbar vertebrae syndrome model rats, and to observe the effect of acupotomology therapy.

METHODSOne hundred and twenty-eight adult SD rats were allocated to 4 groups randomly: normal group, model group, aminoguanidin group and acupotomology treatment group, 32 rats in each group. NOS expression, NO content and injury of the soft tissue in the 3rd lumbar vertebra were observed on the 1st, 3rd, 7th and 14th day after the acupotomology treatment and aminoguanidine intervention.

RESULTS1) Inducible NOS (iNos) activity and NO content in model group was significantly higher (F = 522.860, P < 0.01), in acupotomology group and aminoguanidine group was significantly lower than the model group (FiNOS = 28.894, P < 0.01), and iNOS activity and NO content in all groups was in competence with the condition of soft tissue injuries. 2) Endothelium NOS (eNOS) expression raised in model group and acupotomology group, and achieve peak on the 7th day. There was significant difference between the eNOS expression in acupotomology group and the model group (FeNOS = 3.454, P < 0.05). 3) The expression of neuron NOS (nNOS) in the model group, aminoguanidine group and acupotomology group had no significant (FnNOS = 0.962, P > 0.05).

CONCLUSIONAcupotomology treatment can restrain the development of high content NO, release the inflammatory reaction and injury condition, improve microcirculation, prevent the development of scar tissue of the injured soft tissue, and has significant recovering effectiveness in the soft tissue injured model rats.

Animals ; Disease Models, Animal ; Gene Expression Regulation, Enzymologic ; Guanidines ; therapeutic use ; Lumbar Vertebrae ; drug effects ; metabolism ; pathology ; surgery ; Male ; Muscle, Skeletal ; drug effects ; metabolism ; pathology ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Syndrome ; Time Factors