Objective The gastrointestinal tract of neonate becomes colonized with microorganisms from environment and mother immediately after birth. Strong evidences suggested that early composition of the microflora in neonates play an important role for postnatal development of the immune system. This study was designed to investigate the effect of mode of delivery on the development of gut microflora and characteristics of the stool in breast-fed infants by molecular biology methods. Methods Sixty healthy breast-fed term infants were enrolled in the study(including vaginal delivery group and cesarean section delivery group,each group included 30 infants). Anthropometric measurements and stool study were done at 6-week,8-week,10-week,and 12-week. Fecal samples were subjected to quantitative fluorescence real-time polymerse chain reaction assays for the enumeration of lactobacili,bifidobacteria,and subtype of bifidobacteria(bifidobacterium longum and bifidobacterium breve). Results The numbers of lactobacilli and bifidobacterium longum species were less in cesarean delivery group than those in vaginal delivery group(P ＜ 0.05). Stool mean pH was lower in vaginal delivery group than that in cesarean section delivery group(P ＜ 0.05). There were no differences in stool characteristics and growth between two groups. Conclusions Cesarean section is associated with the decrease of bifidobacteria and lactobacilli in breast-fed term infants.

Aged ; Air Pollutants, Occupational ; Eosinophil Cationic Protein ; blood ; Humans ; Immunoglobulin E ; blood ; Inhalation Exposure ; Male ; Middle Aged ; Silicosis ; blood

OBJECTIVE: To determine the allelic frequency distribution and genetic parameters of nine non-CODIS DNA index systems of the short tandem repeat (STR) loci (D2S1772, D6S1043, D7S3048, D8S1132, D11S2368, D12S391, D13S325, D18S1364, and GATA198B05). METHODS: A total of 353 blood samples were collected, extracted, amplified, and analyzed from unrelated healthy individuals of Han nationality in Hunan Province, China. RESULTS: One hundred and fourteen alleles were observed in the population with corresponding allelic frequencies ranged from 0.001 0 to 0.323 0. For all the nine non-CODIS STR loci, the observed genotypic data showed no significant deviations from the Hardy-Weinberg equilibrium. The Ho, He, PIC, DP, and PE of the studied non-CODIS STR loci ranged from 0.1080 to 0.1950, 0.8050 to 0.8920, 0.7700 to 0.8600, 0.9250 to 0.9660 and 0.6070 to 0.7800, respectively. CONCLUSION Nine non-CODIS STR loci have high degrees of polymorphisms, which may be useful in individual forensic identification and parentage testing in forensic practice.
Alleles ; Asian People/genetics* ; China ; Ethnicity/genetics* ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Male ; Microsatellite Repeats/genetics* ; Polymorphism, Genetic

OBJECTIVETo study the expression of lipoprotein related genes in subchondral bone of early experimental os-teoarthritis, which may play an important role in the pathogenesis of osteoarthritis.

METHODSAnimals are equally divided into two groups: experimental group and control group, both of which contain fifteen rats of similar weight. The right knee joints of experimental group underwent surgery,which involved in both medial collateral ligament(MCL) transaction and medial meniscectomy, while the control group was only carried out with a sham operation. Rats were killed at 1, 2 and 4 weeks postsurgery to obtain the right knee joints. Total RNA of the subchondral bone was extracted,and then hybridized to Agilent Whole Rat Genome Microarray. Differentially expressed genes analysis was used to study the chemokine signaling pathway.

RESULTSApoa5 expression was down-regulated at 1, 2 weeks post-surgery, Apoc2 expression was up-regulated at 1 week after surgery, Apol3 expression was up-regulated at 1 and down-regulated at 4 weeks post-surgery, Lrp1 expression was down-regulated at 1, 2 weeks after surgery. Lrp5 was down-regulated at 2 weeks after surgery. Gpihbp1, Lpl, Tfpi and Vldlr were up-regulated at 1 weeks after surgery. Lrpap1 and RGD1309808 were down-regulated at 4 weeks after surgery.

CONCLUSIONDysregulation of lipoprotein related genes plays an important role in pathogenesis of early experimental osteoarthritis.

Animals ; Disease Models, Animal ; Knee Joint ; metabolism ; Lipoproteins ; genetics ; Male ; Oligonucleotide Array Sequence Analysis ; Osteoarthritis ; genetics ; Rats ; Rats, Sprague-Dawley ; Transcriptome

Objective To analyze the basic clinical characteristics,auxiliary diagnostic indexes,bacterial infection spectrum and drug resistance of child urinary tract infection to provide the basis for the prevention,diagnosis and treatment of child UTI.Methods The clinical data in the inpatients with UTI or complicating UTI in our hospital from January 2012 to March 2014 were collected. The repeated strains were excluded.The differences in the pathogens between the patients with complicated UTI and the patients with non-complicated UTI were comprehensively analyzed.Results The onset peak of child UTI for the first time was 0 -0.5 years old.Escherichia coli ,Klebsiella and Pseudomonas aeruginosa are the major pathogens of child UTI,in which 110 strains were Escherichia coli,accounting for 52.9%.The enterobacteriaceae pathogens of UTI had higher sensitivity to carbapenems antibacterial drugs,with the resistance rate of less than 10%;the resistance rate of others detected antibacterial drugs was more than 20%.Con-clusion UTI in the children inpatients of this area has higher drug resistance rate,in the treatment of child UTI,the antibacterial drugs should be rationally used by combining the bacterial drug resistance situation in the local place and the disease severity in or-der to avoid the aggravation of bacterial drug resistance.

Objective To analyze the etiological factor and genetic feature of a familial hemophagocytic lymphohistiocytosis patient with PRF1 mutation (FHL2) with human herpesvirus 7 (HHV7)infection and its family constellation. Methods Clinical characteristics, laboratory examinations of a FHL2 case with HHV7 infection were reported. HHV1-HHV8 virus DNA was screened by PCR; NK cell function was analyzed by flow cytometry; PRF1 gene mutations were analyzed by PCR and direct sequencing, structure of mutant PRF1 proteins were analyzed using ExPasy and I-TASSER server and genetics pedigree were analyzed. Results The patient's HHV7 viral was detected positive with DNA copy number of 350/106 peripheral nucleated cells. Flow cytometry analysis showed decrease both in proportion of perforin positive NK cells and perforin protein expression. Genetic testing showed PRF1 biallelic heterozygote mutations (c. 503G ＞ A/p. S168N and c. 1177T ＞ C/p. C393R) and pedigree analysis showed they were inherited. The patient was then treated with antivirus therapy, dexamethasone and VP16 therapy, but only achieved partial response. The patient was then followed by human leukocyte antigen 10/10 allele identical nonconsanguinity allogeneic hematopoietic stem cell transplantations (allo-HSCT) and soon the successful implantation of donor hematopoietic cells and persistent recovery was achieved. The patient was now surviving without recurrence for 9 months after allo-HSCT. Conclusions FHL is prone to be misdiagnosed as lymphoma. Genetic analysis of related gene mutation and herpes simplex virus detection will help in early and accurate diagnosis. Allo-HSCT is a fundamental treatment of FHL.

Objective To observe the clinical efficacy of acupoint thread embedding in preventing urinary retention after cervical cancer surgery.Method Fifty eligible patients were randomized into a treatment group and a control group, 25 cases each. The control group was given catheterization after gynecological surgery; the treatment group was intervened by thread embedding at Zusanli (ST36), Sanyinjiao (SP6), Yinlingquan (SP9), Guanyuan (CV4), and Shenshu (BL23) on the 7th day after the surgery in addition to the basic intervention given to the control group. The indwelling catheter was removed on the 14th day after the surgery, and the bladder function, urination, and residual urine volume were compared between the two groups.Result After the treatment, the bladder function in the treatment group was significantly better than that in the control group (P<0.05); the markedly effective rate of spontaneous micturition was 92.0% in the treatment group versus 72.0% in the control group, and the difference was statistically significant (P<0.05).Conclusion Early intervention with acupoint thread embedding after cervical cancer surgery can effectively decrease the incidence of post-surgery urinary retention, and it's easy-to-operation and worth application in clinic.

Based on the outpatient interview and literature review,the initial framework of the outpatient experience of human caring scale was formed with 9 dimensions of outpatient process.The research aim was to improve the scale by Delphi method.Sixteen experts in medical management,human caring or medical education were invited to evaluate the importance of the dimensions and items of the scale and provided some expertise via filling out the Delphi consultation questionnaires twice in the consulting round.In the first round,the recovery rate showing the experts' positivity was 80％;the coefficient of reliability (Cr) ascertaining the authority of the evaluation was 0.92;the mean and full mark ratios responding the concentration of the evaluation were 2.88-4.94 and 6.25％-93.75％ respectively;the coefficients of variation (CV) and the Kendall's W determining the concordance of the evaluation were 5.06％-52.15％ and 0.21-0.24 respectively.In the second round,the recovery rate was 93.75％;the Cr was 0.93;the mean was 3.93-4.93;the full mark ratios were 26.67％-93.33％;the Kendall's W was 0.14-0.31,the CV was 5.25％-23.61％.Via the two-round Delphi study,the scale that included 10 dimensions and 61 items has been improved.Ten dimensions are pre-hospital medical service,guidance,registration,waiting,diagnosis & treatment,paying,inspection & assay,medicine receiving,therapy/injection/transfusion and global evaluation.It was concluded that Chinese scholars have paid high attention to human caring and outpatient experience.The experts have given high agreements about the dimensions which were established with Chinese outpatient process.The dimensions are different from the similar researches about outpatient experience study.In the future,it is necessary to survey the outpatients to test the construct validity,internal consistency reliability and others of the scale to improve the scale.

Human Zona Pellucida(ZP), which is a complex matrix surrounding oocytes,is comprised of three immunologically distinct glycoproteins(hZP1, hZP2 and hZP3). Because hZP3 possesses the sperm receptor activity and the acrosome-inducing activity, it has long been used as a candidate antigen to develop an immunocontraceptive vaccine. However, a large amount of native hZP3 protein is unavailable. It is an effective way to express hZP3 protein directly in vitro. Nevertheless, it had been reported that the rhZP3 protein produced in Pichia pastoris was not secreted but accumulated in the cells and could only be purified after being solubilized by strong denaturants. More unfortunately, after purification the final product required 6mol/L urea to maintain solubility. An improved project was advanced with the aim to express secreted and soluble rhZP3 protein in yeast. In this study, the fragment of hZP3 cDNA coding for aa 23 - 408, which the N-terminal leader was removed and most of the C-terminal transmembrane-like domain was reserved, was amplified by two PCR primers including EcoR I and Not I sites respectively and a His6 codon cassette was added to 5'-terminal. The hZP3 insert was incorporated into expression vector pPIC9K. The resulting recombinant yeast expression vector was designated pPIC9K-rhZP3. Linearized pPIC9K-rhZP3 was transformed into Pichia pastoris. After G418 selection, the recombinant Pichia pastoris strains were identified by PCR and the rhZP3 was expressed following the manufacturer' s protocol. Following induction with methanol, the rhZP3 protein was secreted and dissolved into the culture supernatant. SDS-PAGE and Western blot analyses showed that the apparent molecular weight of the expressed rhPZ3 proteins in yeast was smaller and a little size heterogeneity than native ones; after purified with Ni-chelating affinity chromatography, the final product's apparent molecular weight was about 32 - 34KD and their yield more than 20mg/L. We supposed that the C-terminal transmembrane-like domain be useful for secretion of rhZP3 into the culture supernatant and the expressed rhZP3 protein be incompletely digested by proteinases of Pichia into shorter fragments which all were glycosylated inhomogeneously. Fortunately, the fragments of rhZP3 protein can be recognized in Western blot by the polyclonal antibodies to porcine ZP3 which has showed a cross-reactivity with human ZP in vitro. It will be expected that the rhZP3 protein expressed in Pichia pastoris not only has immunogencity, say, it can rise antibodies in vivo to prevent spermatozoa-ovum binding, but also does not contain ovarian factors that might be the cause of undesired side effects, e.g. ovaritis and can be used as a safe immunogen in human antifertility vaccine research.
Blotting, Western ; Chromatography, Affinity ; Egg Proteins ; genetics ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Genetic Vectors ; genetics ; Humans ; Membrane Glycoproteins ; genetics ; metabolism ; Pichia ; genetics ; metabolism ; Polymerase Chain Reaction ; Receptors, Cell Surface ; genetics ; metabolism ; Recombinant Proteins ; genetics ; isolation & purification ; metabolism ; Zona Pellucida Glycoproteins

OBJECTIVETo investigate the immunohistochemical expression of isocitrate dehydrogenase 1 gene (IDH1) R132H in glioma and its diagnostic utility.

METHODSImmunohistochemical study of IDH1R132H expression was performed on formalin-fixed paraffin-embedded tissue samples of 75 gliomas, including 33 cases of grade II, 20 cases of grade III and 22 cases of grade IV tumors. Six cases of pilocytic astrocytoma and 12 cases of gliosis were used as controls.

RESULTSNineteen in 33 cases of grade II (57.6%), 8 in 20 cases of grade III (40.0%), 6 in 22 cases of grade IV (27.3%) showed positive cytoplasmic staining of IDH1R132H. Scattered invasive glioma cells at the tumor periphery also expressed IDH1R132H. Gliomas involving the frontal lobe showed more strong IDH1R132H staining. In contrast, none of the pilocytic astrocytomas and gliosis showed IDH1R132H staining. Moreover, the rate of p53 immunopositivities were 42.4% (14/33) in grade II, 65.0% (13/20) in grade III and 77.3% (17/22) in grade IV gliomas. There were no statistic correlations between expression of IDH1R132H and p53.

CONCLUSIONIDH1R132H tends to express preferentially in low-grade gliomas, and it thus may serve as a valuable marker in distinguishing low grade gliomas from gliosis.

Adolescent ; Adult ; Aged ; Astrocytoma ; metabolism ; pathology ; Brain Neoplasms ; metabolism ; pathology ; Child ; Diagnosis, Differential ; Female ; Glioma ; metabolism ; pathology ; Gliosis ; metabolism ; pathology ; Humans ; Isocitrate Dehydrogenase ; genetics ; metabolism ; Male ; Middle Aged ; Mutation ; Tumor Suppressor Protein p53 ; metabolism ; Young Adult