Objective:To explore the profile and gene functional changes of gut microbiota (GM) in women with postmenopausal osteoporosis (PMOP) in Northwest China, and the correlations between GM and bone mineral density (BMD).Methods:From November 2018 to October 2019, postmenopausal women were screened on their initial visits to our hospital, and 24 new osteoporosis (OP) patients, 30 new osteopenia patients and nine negative controls were recruited. Fecal samples were collected for GM DNA extraction, and Illumina platforms were used for high-throughput sequencing of 16S rRNA and metagenome. Species annotation, GM profile and gene functions were viewed and analyzed.Results:GM profiles were significatly different in different groups, and the LDA scores of Peptostreptococcaceae, Romboutsia, unidentified Clostridiales, Megamonas, Erysipelatoclostridium, Klebsiella and Erysipelatoclostridium ramosum were more than 3 in OP group. Metagenomic sequencing analysis indicated that gene numbers were positively correlated with BMD, and metabolism, carbohydrate metabolism, starch and sucrose metabolism and oxidative phosphorylation were negatively correlated with BMD. Receiver operating characteristic curve(ROC) showed that carbohydrate metabolism, starch and sucrose metabolism, amino sugar and nucleotide sugar metabolism, oxidative phosphorylation, respectively, could identify OP with preferable sensitivity and specificity (areas under curve were 0.70, 0.72, 0.73 and 0.75, respectively). Conclusions:High-throughput sequencing had great potential for GM analysis of postmenopausal women with OP, providing evidence of the correlations between GM and BMD.

Objective: To investigate the recovery of splenomegaly and hypersplenism after liver transplantation in children and explore the necessity of splenomegaly management before and during liver transplantation. Method: The data of 22 children who were underwent liver transplantation with preoperative splenomegaly and hypersplenism who were not treated with splenomegaly before and during operation and with no recurrence of portal hypertension during postoperative follow-up in Department of Hepatobiliary Surgery, Children’s Hospital of Chongqing Medical University from December 2008 to January 2019 were collected. There were 13 male patients and 9 female patients with a median age of 6.5 months. The changes of erythrocyte, platelet, white blood cell and spleen length and thickness were analyzed by paired t-test before and at the 1st, 3rd and 6th months after surgery. Correlation analysis was performed on the change of spleen long diameter and thickness product and blood cell recovery. Results: There were 21 cases of red blood cell decreased before surgery, with a mean value of(3.1±0.5)×10¹²/L, and(3.7±0.7)×10¹²/L, (4.6±0.6)×10¹²/L and (4.3±0.5)×10¹²/L at the 1st, 3rd and 6th months after operation, respectively. The differences between the preoperative and postoperative groups were statistically significant (P<0.05). There were 7 cases of thrombocytopenia before operation, with an average of (70.0±17.0)×10⁹/L, and (191.0±129.0)×10⁹/L, (156.0±79.0)×10⁹/L and (167.0±63.0)×10⁹/L at the 1st, 3rd and 6th months after operation, respectively. The differences between the first, third, and sixth months after surgery were significant compared with that before surgery (P<0.05). Leukocyte count decreased in 5 cases with an average value of (3.30±0.56)×10⁹/L before surgery, (7.5±4.4)×10⁹/L, (7.4±1.4)×10⁹/L, (5.1± 2.5)×10⁹/L at the 1st, 3rd and 6th months after operation, respectively. The differences between the two groups were significant (P<0.05). The average value of preoperative spleen length times thickness was (35.7±12.9) cm² and at the 1st, 3rd, 6th month after surgery it was (26.2±8.1)cm², (25.2±13.4)cm², (27.2±7.4)cm², respectively. The differences between the first, third months after surgery were significant compared with that before surgery (P<0.05). The white blood cell and platelet counts of children before and after surgery were correlated with spleen size negatively (correlation coefficients were -0.902, -0.933, respectively), and the differences were statistically significant (P<0.05). Conclusions In the early stage of liver transplantation, the size of spleen retracts to varying degrees, and hypersplenism can be alleviated continuously. Therefore, it is unnecessary to treat splenomegaly and hypersplenism before and during liver transplantation.

Objective: To analyze the type of malocclusion which was more likely to cause undesirable orthodontic results and the reasons for orthodontic re-treatment. Methods: In this study, 202 patients who received orthodontic retreatment in School of Stomatology, China Medical University from 2004 to 2016 were collected. Forty-seven patients were teenagers under 18 years old, and 155 patients were adults over 18 years old. The chief complaint of all patients was analyzed. Seven orthodontic specialists were invited to analyze the reasons caused the orthodontic re-treatment according to the treatment records, plaster models, photographs, panoramic radiographs and lateral cephalograms. They were offered an evaluation form to fill in by their judgement. The reasons for orthodontic re-treatment and the problems related to previous orthodontic treatment were analyzed. Results: In 202 patients, the chief complaints were protrusion of the teeth [30.2% (61/202)], protrusion of the mandible [20.8% (42/202)], and irregularity of the teeth [17.8% (36/202)]. The proportion of patients chosen undesirable treatment design was 68.3% (138/202). The proportion of patient having unusual growth tendency was 24.3% (49/202). The proportion of patients with poor compliance during treatment and retention was 24.3% (49/202). Conclusions In this study, the protrusion of the teeth, the protrusion of the mandible and the dental crowding were the main chief complaints for orthodontic re-treatment. Undesirable treatment design and unfavorable growth tendency were the main reasons for orthodontic re-treatment. Poor compliance of patients during the period of treatment or retention was an important reason for the patient to seek orthodontic re-treatment.

Objective: To analyze factors affecting stability after fixed orthodontic treatment. Methods: Five hundred and forty-four patients who had finished fixed orthodontic treatment more than two years in First Department of Orthodontics, China Medical University from January, 2000 to December, 2017 were investigated and the rate of regular revisit was counted. The data of 288 patients were successfully collected by calling or sending text messages and the rate of standard use of retainers as prescribed (patients wore retainers for 24 months or longer) was counted. According to the variation of peer assessment rating (PAR) index at the start of retention and the time the data collected, the patients were divided into relapse group (variation of PAR index >5) and non-relapse group (variation of PAR index ≤5). Difference significance analysis and multiple-factor logistic regression analysis were used. Sixty patients wearing retainers well were collected and the curative effects of Hawley retainer and vacuum formed retainer (VFR) were compared, which included overbite, overjet, maxillary irregularity index, mandibular irregularity index, width betwenn canine and width between first molar. Results: The rate of regular revisit was 41.0% (223/544). Two hundred and one of 288 patients (69.8%) who were visited successfully had regular revisit, and 60.4%(174/288) of the patients wore retainers well; 30.2% (87/288) of the patients who were visited successfully didn't have regular revisit, and 10.4% (30/288) of the patients wore retainers well. Difference significance analysis showed that there was highly significant difference between relapse group and non-relapse group in the type of retainer and duration of retention (P<0.01). One hundred and forty of 224 patients (62.5%) in non-relapse group and 37.5% (24/64) of the patients in relapse group used VFR, and the duration of retention in non-relapse group was significantly longer than that in relapse group (P<0.01). Multiple-factor Logistic regression analysis showed that wearing Hawley retainers (OR=3.067, P<0.05) was the risk factor influencing relapse. The duration of retention (OR=0.832, P<0.01) was the protective factor influencing relapse. Independent-sample t test indicated that the variations of maxillary [(0.82±0.36) mm] and mandibular [(1.05±0.22) mm] irregularity index in Hawley retainer group were larger than maxillary [(0.64±0.29) mm] and mandibular [(0.72±0.35) mm)] irregularity index in VFR group, respectively. The differences between the two groups were significant (P<0.05). Conclusions Duration of retention was implicated in stability after orthodontic treatment. VFR had better effect in the aspects of irregularity index than Hawley retainer.

OBJECTIVETo explore the subcellular localization of ataxin-3 and the effect of polyglutamine (polyQ) expansion mutation on the morphology of mitochondrion, golgi apparatus and endoplasmic reticulum.

METHODSTransient transfection was employed to build cell models expressing wild-type or mutant ataxin-3 proteins. Indirect immunofluorescence was applied to identify markers of organelle membrane. The results were observed under a laser scanning confocal microscope.

RESULTSNo co-localization was observed for ataxin-3 protein and mitochondrial marker TOM20, but the percentage of cells with mitochondrial fragmentation has increased in cells expressing mutant ataxin-3 (P<0.05). No co-localization was observed for ataxin-3 protein and golgi marker GM130, and mutant ataxin-3 did not cause golgi fragmentation. Wide type and polyQ-expanded ataxin-3 both showed partial co-localization with ER marker calnexin. The latter showed more overlap with calnexin, and the overlapping signals were mostly located in the places where aggregates were situated.

CONCLUSIONPolyQ-expanded ataxin-3 protein may indirectly affect the integrity of mitochondria, but may cause no effect on the structure and functions of golgi apparatus. Endoplasmic reticulum may be another place where extended ataxin-3 protein can induce cytotoxicity in addition to the nucleus.

Ataxin-3 ; Cytoplasm ; genetics ; metabolism ; Endoplasmic Reticulum ; genetics ; metabolism ; HeLa Cells ; Humans ; Machado-Joseph Disease ; genetics ; metabolism ; Mitochondria ; genetics ; metabolism ; Nerve Tissue Proteins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Protein Transport ; Repressor Proteins ; genetics ; metabolism

Objective To determine the prevalence of penicillinase-producing Neisseria gonorrhoeae(PPNG) and the distribution of blaTEM-135 gene variants in PPNG at several gonococcal antimicrobial surveillance sites in China, to compare N. gonorrhoeae multi-antigen sequence typing(NG-MAST)types of PPNG and its blaTEM-135 gene variants, and to assess the difference and association in NG-MAST types of blaTEM-135 gene variants among different regions. Methods A total of 572 N. gonorrhoeae isolates were collected at 6 gonococcal antimicrobial surveillance sites from Jiangsu, Shanghai, Zhejiang, Tianjin, Guangdong and Guangxi in 2012. After isolation, purification, and identification, cefalotin paper discs were used for detection of PPNG. DNA was extracted by QIAxtractor DX kits after cultivation of the PPNG strains. Then, mismatch amplification mutation assay (MAMA) PCR was performed to identify blaTEM-135 variants, and NG-MAST analysis to determine N. gonorrhoeae genotypes. Results Among the 572 N. gonorrhoeae strains, 38.1%(218/572) were identified as PPNG, and of the PPNG strains, 52.3% (114/218) were blaTEM-135 variants. The detection rate of PPNG at these surveillance sites from high to low was as follows: 51.7% (45/87, Zhejiang), 45.6%(36/79, Shanghai), 38.0% (78/205, Guangdong), 37.5% (12/32, Guangxi), 31.2% (24/77, Jiangsu) and 25.0%(23/92, Tianjin), and that of blaTEM-135 variants was as follows: 68.9%(31/45, Zhejiang), 58.3%(14/24, Jiangsu), 50.0%(39/78, Guangdong), 47.2%(17/36, Shanghai), 39.1%(9/23, Tianjin)and 33.3%(4/12, Guangxi). NG-MAST analysis showed that the ST2318, ST1768, ST1866, ST1053 and ST8726 types predominated among these bla TEM-135 variants, and a strong correlation was found between blaTEM-135 variants and some NG-MAST types, such as ST1768, ST1053 and ST8726 types. The distribution of NG-MAST types was significantly different between the surveillance site in Tianjin (in the Northern part of China) and the other sites (in the Southern part of China), but highly similar among the surveillance sites in Jiangsu, Zhejiang and Shanghai regions. Conclusions There is a high prevalence of PPNG and its blaTEM-135 variants at several gonococcal antimicrobial surveillance sites in China, with significant differences in NG-MAST genotype distribution of PPNG and its blaTEM-135 variants among different regions.

Objective To study the application effect of clinical pathway teaching in respiratory medicine. Methods Seventy clinical medical students of our department during 2007 to 2009 were randomly divided into two groups. The control group (35 students) was treated by traditional teaching ways, while the experimental group(35 students) was treated by clinical pathway teaching ways, with 5 to 6 students forming a small group. Teachers provided one copy of the CP version to each person in advance. Then progressive questions and discussions were conducted according to the diagnosis, dif-ferential diagnosis and treatment of CP. After its implementation for a certain time, students were co-mprehensively assessed by the practice examination and questionnaire and the statistical analysis was performed by using SPSS 17.0 version statistics software. Results The experimental group's total aver-age score was (90.00±4.00) points, while the control group's total average score was (76.00±7.20) points. There was significant difference between the two groups(P=0.001). The effect of these two kinds of teaching was evaluated and compared in stimulating interest in learning (P=0.002), improving the analytical ability(P=0.004),improving self-study ability(P=0.001), deepening the under-standing of the basic concepts(P=0.112), improving the innovation ability (P=0.005), improving the efficiency of learning(P=0.034), improving clinical comprehensive ability(P=0.016), and improving the ability of language expression(P=0.000), showing that teaching method of clinical pathway could significantly improve clinical teaching effect, and there was statistically significant difference between them. Conclusion Clinical pathway teaching has obvious advantages in cultivating students' diagnostic thinking and clin-ical ability to solve practical problems, and therefore it has better clinical teaching effect than the tra-ditional teaching method.

OBJECTIVETo explore the relationship between the syndrome types in traditional Chinese medicine (TCM) and serum HBV DNA load in chronic HBV carriers positive for HBeAg.

METHODSAccording to the TCM syndrome types, 185 chronic HBV carriers with HBeAg positivity were classified into single syndrome group (liver Qi depression, kidney Qi deficiency, spleen Qi deficiency, and kidney Yang deficiency), compound syndrome group, and unidentifiable syndrome group; based on the nature of the condition in TCM terms, the patients were classified into excess syndrome group, deficiency syndrome group and comorbidity syndrome group. The serum HBV DNA levels in these cases were analyzed in relation to the TCM syndrome types and disease nature.

RESULTSHBV DNA levels showed no significant difference among the patients with single syndrome, compound syndromes and unidentifiable syndrome (F=0.910, P=0.404), nor among the patients with the 5 different single TCM syndromes (χ²=4.672, P=0.323) or those with different disease nature (F=0.631, P=0.596).

CONCLUSIONSerum HBV DNA level can not be considered as the evidence for syndrome differentiation in chronic HBV carriers with positive HBeAg.

Adolescent ; Adult ; Carrier State ; blood ; diagnosis ; Child ; DNA, Viral ; blood ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; blood ; diagnosis ; virology ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Young Adult

We developed a new method for soluble expression of phage-displayed scFv antibody specific for zebrafish vitellogenin. The scFv antibody F5 could bind zebrafish vitellogenin specifically in phage-displayed form but not soluble form. The gene of scFv antibody F5 was cloned into vector pET 32a and transferred into Escherichia coli ori DE3. With inducible expression, soluble scFv antibody 32a-F5 was obtained successfully and could also specifically bind to zebrafish vitellogenin. The insoluble expression of phage-displayed scFv antibody was a common problem in the practical use of phage display. This study offered a feasible way to express soluble scFv antibodies with biological activity.
Amino Acid Sequence ; Animals ; Antibody Specificity ; Base Sequence ; Cell Surface Display Techniques ; Escherichia coli ; genetics ; metabolism ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Single-Chain Antibodies ; biosynthesis ; genetics ; immunology ; Solubility ; Vitellogenins ; immunology ; Zebrafish ; immunology ; metabolism

Antibodies are immunoglobulins specifically introduced by immunity response of high animals, with the responsibility for recognising and cleaning out specific antigens. Antibody is not only a powerful weapon against pathogen invasion in the organism, but also a tool for specific molecular recognition used in basic scientific research. The diversity of antibody molecules resulted in the concept of antibody library; each individual animal is a natural antibody library. In the post-genome era, in order to fit various "omics", especially for proteomics requirement of high throughput technology, some gene engineering antibody libraries and antibody alternative libraries have been constructed based on phage display technology. Yet, more and more in vitro display systems such as ribosome display, mRNA display have been used for antibody library study, and that present more advantages than phage display. This mini review outlines the genesis, development and application prospect of antibody libraries according to the published reviews and research articles, and offers up to date development and application prospect of antibody library technology.
Animals ; Antibodies ; genetics ; physiology ; Antibody Diversity ; genetics ; Antibody Specificity ; Combinatorial Chemistry Techniques ; Gene Library ; Humans ; Peptide Library