Objective　To explore the molecular epidemiological characteristics of local Mycobacterium tuberculosis in Chongqing, China using genotyping methods and establish a suitable genotyping system tuberculosis genotyping in the region. Methods A total of 268 isolates collected from the sputum of tuberculosis patients at two national drug resistance monitoring sites in Chongqing from 2019 to 2021. The DNA of these isolates was extracted, and genotyping of the isolates was performed using VTNR 24 locus typing method. Epidemiological investigations were conducted on clustered isolates, and the clustering rates of isolates from the two surveillance sites were compared to analyze their transmission differences. Results In the two regions, the Hunter-Gaston discriminatory index (HGI) in Fengjie County ranged from 0 to 0.85, with 10 loci having an HGI above 0.6, 11 loci with an HGI between 0.3 and 0.6, and 3 loci with an HGI below 0.3; the highest resolution site was MIRU31, while the lowest resolution site was MIRU24. The HGI index in Fengdu County ranged from 0 to 0.81, with 12 loci having an HGI above 0.6, 9 loci with an HGI between 0.3 and 0.6, and 3 loci with an HGI below 0.3; the highest resolution site was MIRU26, while the lowest resolution site was MIRU23. In the clustering analysis, among the 140 isolates from Fengdu County, 10 samples clustered into 4 clusters, with the largest cluster consisting of 4 samples, resulting in a clustering rate of 4.1%. Among the 128 isolates in Fengjie County, 10 samples clustered into 5 clusters, with a clustering rate of 4.2%. Conclusion In Fengdu County and Fengjie County of Chongqing, the main population of the disease show independent genotypes, indicating a low recent transmission rate. The incidence of patients is mainly endogenous recurrence, that is, latent infection develops into active tuberculosis. While conducting standardized management of tuberculosis patients, prevention and control agencies should also increase the screening of latent infection at two monitoring points and carry out preventive treatment of latent infection, which is the key to reduce the incidence of tuberculosis in the two places.

Objective:To explore whether microRNA (miRNA) -181b-5p inhibits the proliferation and invasion of cutaneous melanoma cells by targeting pleckstrin (PLEK) .Methods:Bioinformatics methods were used to analyze cutaneous melanoma-associated core genes; dual-luciferase reporter assay was performed to verify the targeted interaction between miRNA-181b-5p and PLEK. Oligo RNA and small interfering RNA (siRNA) were used to regulate the expression of miRNA-181b-5p and PLEK in A375 cells respectively in this experiment, and A375 cells were divided into the following groups in detail: mimic negative control group, miRNA-181b-5p mimic group, inhibitor negative control group, miRNA-181b-5p inhibitor group, PLEK siRNA group, siRNA negative control group, miRNA-181b-5p inhibitor + control siRNA co-transfection group and miRNA-181b-5p inhibitor + PLEK siRNA3 co-transfection group. After 48-hour treatment, qPCR was performed to determine the mRNA expression of miRNA-181b-5p and PLEK in A375 cells, Western blot analysis to determine the PLEK protein expression, and Transwell assay to assess the invasive ability of A375 cells; after additional 24-96 hours of culture, cell counting kit-8 (CCK8) assay was conducted to assess the proliferative ability of A375 cells.Results:PLEK was the core gene for cutaneous melanoma. PLEK expression in the cutaneous melanoma in situ tissues was significantly higher than that in the paracancerous tissues ( P = 0.031) , but lower than that in the metastatic tissues ( P = 0.001) . Compared with human epidermal melanocytes HEMa-LP, the mRNA and protein expression of PLEK significantly increased in A375 cells (mRNA: 3.884 ± 0.156 vs. 0.997 ± 0.010, t = 18.48, P < 0.001; protein: 2.840 ± 0.301 vs. 1.029 ± 0.094, t = 5.47, P = 0.005) , but the miRNA-181b-5p expression significantly decreased in A375 cells (0.333 ± 0.042 vs. 0.967 ± 0.069, t = 7.83, P = 0.001) . Dual-luciferase reporter assay showed targeted binding of miRNA-181b-5p to PLEK. Compared with the mimic negative control group, the miRNA-181b-5p mimic group showed significantly decreased survival rate of A375 cells (48 hours: t = 7.96, P = 0.015; 72 hours: t = 7.50, P = 0.002; 96 hours: t = 7.96, P = 0.001) , and significantly decreased invasive ability of A375 cells ( t = 5.07, P = 0.007) ; on the contrary, the survival rate and invasive ability of A375 cells were significantly higher in the miRNA-181b-5p inhibitor group than in the inhibitor negative control group (survival rate: 24 hours, t =5.38, P = 0.013; 48 hours, t = 5.36, P = 0.013; 72 hours, t =7.63, P = 0.005; 96 hours, t = 5.99, P = 0.004; invasive ability: t = 7.24, P = 0.002) ; compared with the siRNA negative control group, the proliferative and invasive ability of A375 cells significantly decreased in the PLEK siRNA group (proliferative ability: 48, 72, 96 hours, P = 0.015, 0.011, 0.001, respectively; invasive ability: t = 4.93, P = 0.008) ; compared with the miRNA-181b-5p inhibitor + control siRNA co-transfection group, the miRNA-181b-5p inhibitor + PLEK siRNA co-transfection group showed significantly decreased proliferation rate and invasive ability of A375 cells (proliferation rate: 24, 48, 72, 96 hours, P = 0.042, 0.042, 0.037, 0.017, respectively; invasive ability: t = 8.52, P = 0.001) . Conclusion:miRNA-181b-5p can inhibit the proliferation and invasion of cutaneous melanoma A375 cells, likely by down-regulating the PLEK expression.

Objective:By establishing the reference intervals of free thyroxine (FT 4), thyroid stimulating hormone (TSH) and thyroid peroxidase antibody (TPOAb) of pregnant women in different pregnancy in Yangzhou, and analyzing the dynamic trends of each indicator, so as to provide a basis for timely and accurate diagnosis of thyroid disease during pregnancy, and promote prenatal and postnatal care. Methods:Clinical data of 3 726 healthy early (1 747 cases), middle (1 481 cases), and late (498 cases) pregnant women were collected from the Department of Perinatal and Health Care in Yangzhou Women and Children Hospital, the Affiliated Hospital of Yangzhou University Medical College from October 2017 to October 2018. At the same time, data of 407 non-pregnant women in the same period were collected as normal controls. The levels of serum FT 4, TSH and TPOAb in each stage of pregnancy were detected by Beckman automatic chemiluminescence analyzer. The reference interval was established by using the 95% reference value of the bilateral limit, and the differences of early, middle and late pregnancy were compared. Results:There were significant differences in FT 4 levels between early, middle and late stages of pregnancy ( H = 82.56, P < 0.01), with the early stage higher than the middle stage ( P < 0.01) and the middle stage higher than the late stage( P < 0.01). The difference of TSH levels was statistically significant ( H = 91.27, P < 0.01), in which the early stage was lower than the middle stage ( P < 0.01), and the middle stage was lower than the late stage ( P < 0.01). There was a statistically significant difference in TPOAb levels ( H = 30.36, P < 0.01). There was no significant difference between the early stage and the middle stage ( P > 0.05), and the early and middle stages were higher than the late stage ( P < 0.01). The reference intervals of thyroid function index in different pregnancies were, early pregnancy: FT 4 8.28 - 15.66 pmol/L, TSH 0.11 - 4.23 mU/L, TPOAb 0.10 - 16.46 U/ml; middle pregnancy: FT 4 7.38 - 14.36 pmol/L, TSH 0.13 - 4.67 mU/L, TPOAb 0.10 - 18.97 U/ml; and late pregnancy: FT 4 6.33 - 11.39 pmol/L, TSH 0.40 - 3.96 mU/L, TPOAb 0.10 - 6.17 U/ml. Conclusions:There are significant differences in serum thyroid function indicators in different pregnant women. Establishing reference intervals of thyroid function indicators in different stages of pregnancy have important clinical significance for diagnosis of thyroid disease and eugenics.

Objective: To screen and identify serum protein biomarkers for the differential diagnosis between ischemic colitis (IC) and ulcerative colitis (UC) by tandem mass tag (TMT) combined with liquid chromatography/tandem mass spectrometry (LC-MS/MS). Methods: From January 2018 to January 2019, at the First Affiliated Hospital of School of Medicine of Zhejiang University, patients with UC or IC, and health controls, each 10 cases, were enrolled into UC group, IC group and normal control (NC) group, respectively. Fasting serum samples of all the subjects were collected. After removal of high-abundance protein, followed by proteolysis, peptide labeling and fractionating, the samples were then processed by mass spectrometry. The protein with TMT data of three groups was obtained and protein with TMT value 0 were removed. Heat map of protein was constructed. The differential protein was defined as the protein fold change over 1.5 or less than 0.67. The Reactome database was used to cluster the pathways of differential proteins among groups. Statistical methods included t test, hypergeometry test and corrected by BH multiple test. Results: A total of 357 serum proteins were identified by proteomic profiling. There were 27 differential proteins between the IC group and the NC group, including six up-regulated proteins and 21 down-regulated proteins. There were 228 differential proteins between the UC group and the NC group, including 75 up-regulated proteins and 153 down-regulated proteins. There were 49 differential proteins between UC group and IC group, including 22 up-regulated proteins and 27 down-regulated proteins. In the comparison of differential proteins between the NC group, IC group and UC group, only the expression of fibrin 3 was statistically significant (the fold change between UC and NC, between UC and IC, between IC and NC were 0.24, 0.46 and 0.53, respectively; t=-5.089, -7.298 and -3.919, all P<0.01). The results of pathway cluster analysis showed that in the comparison of differential proteins between IC group and NC group, only the platelet degranulation pathway was enriched, and 10 proteins were involved in this pathway (P<0.01). In the comparison of differential proteins between UC group and NC group, there were 58 pathways enriched, of which 38 proteins were involved in the platelet degranulation pathway, 16 proteins were involved in the initial complement trigger pathway, 13 proteins were involved in the complement cascade pathway, and 11 proteins were involved in antibody-mediated complement activation pathway (all P<0.01). In the comparison of differential proteins between UC group and IC group, three different pathways were obtained. Among them, nine proteins were involved in the platelet degranulation pathway, seven proteins were involved in the initial complement trigger pathway, and five proteins were involved in the complement cascade pathway (all P<0.01). Conclusions The difference in serum proteome between IC patients and UC patients was significant, and the differential proteins are mainly involved in platelet activation and complement activation. The candidate proteins identified in this study may be used as biomarkers for the differential diagnosis of UC and IC in the future.

Objective To explore the diagnostic model and clinical application value of serum proteomic fingerprint in inflammatory bowel disease (IBD) .Methods Serum proteome profiles of 72 IBD patients (54 Crohn′s disease (CD) and 18 ulcerative colitis (UC) and 44 healthy controls were analyzed by the weak cation exchange (WCX) beads combined matrix‐assisted laser desorption/ionization time of flight mass spectrometry (MALDI‐TOF‐MS ) technique . Among three groups , every two groups were compared .Wilcoxon rank sum test was used to screen out the peaks of difference expressed protein (P<0 .05) .Genetic algorithm combining with support vector machine (SVM ) was utilized to select the best diagnostic model .The predictive effects of this model was evaluated by leave one out method (LOO ) . Results The 10 most discriminating protein peaks were screened out between CD group and healthy control group , between UC group and healthy control group , between CD group and UC group . A diagnostic model established with four protein peaks ,the mass‐to‐charge ratio (M /Z ) of them was 3 275 .29 ,4 963 .91 ,4 980 .53 and 5 336 .90 ,could better distinguish CD and healthy controls .The specificity was 97 .7% ,and the sensitivity was 92 .6% in CD diagnosis .A diagnostic model established with four protein peaks ,the M /Z of them was 2 272 .41 ,2 660 .42 ,3 029 .77 and 5 002 .78 ,could better distinguish UC and healthy controls .The specificity was 100 .0% ,and the sensitivity was 94 .4% .A specificity was 50 .0% and sensitivity was 88 .9% in CD diagnosis with the diagnostic model of six protein peaks and the M /Z of them was 2 082 .63 ,2 210 .64 ,4 039 .02 ,4 298 .30 ,4 978 .03 ,5 002 .22 .Conclusion The diagnostic model of serum difference expressed protein in CD and UC is established by MALDI‐TOF‐MS technique and genetic algorithm combining with SVM ,which has high diagnostic value in IBD .

Aim To observe the effects of effective fraction of Epimedium,Astragalus,Radix Puerariae on behavioral and pathological changes in a transgenic mouse model of Alzheimer’s disease.Methods Six-month-old APPswe /PS1 ΔE9 transgenic mice were ran-domly divided into 2 groups:model group and effective fraction group,1 0 mice each group.The mice in the effective fraction group were treated with the effective fraction of Astragalus,Radix Puerariae,Epimedium compound for 8 weeks.The C57BL/6J mice were used as negative control group.After 8 weeks,the learning and memory function were measured by Morris water maze,the pathological changes in brain tissue were ob-served by Modified Bielschowsky staining and Nissl 's staining.Results During place navigation trial,the escape latency in the APPswe /PS1 ΔE9 double transgenic model mice was longer than those of the mice of C57 (P <0.05),the escape latency in the mice of effec-tive fraction group was significantly reduced than those of the mice in model group (P <0.05).During spatial probe trail,the platform-crossing times in the APPswe /PS1 ΔE9 double transgenic mice were different from the mice of C57 (P <0.05),the platform-crossing times in the mice of effective fraction group were significantly increased than those of the mice in model group (P <0.05).The average swimming velocity in the APPswe /PS1 ΔE9 double transgenic model mice was increased than that of mice of C57 (P <0.05 ),there was no significant difference between effective fraction group and model group (P > 0.05 ). The Modified Bielschowsky staining shows that the neuron fibers of the cerebral cortex of APPswe /PS1 ΔE9 double transgenic mice were enlarged,swelling,and dense.There were senile plaques and nerve fiber tangles in the cerebral cortex of APPswe /PS1 ΔE9 double transgenic mice.The neuron fibers of mice in the effective fraction group were relieved;there was a small amount of senile plaque.The Nissl’s staining shows that the neurons of the cerebral cortex of APPswe /PS1 ΔE9 mice were edema, the number of cells were decreased.The mice in the effective fraction group were free of the disease.Con-clusion The double transgenic APPswe /PS1 ΔE9 mice of AD can simulate the specific pathogenesis of AD, which may be the efficient experimental animal model. The effective fraction of epimedium,astragalus and ra-dix puerariae may have a neuroprotective effect against AD via improving the learning and memory ability,and reduce the cerebral cortex nerve fiber tangles,senile plaques and neurons edema changes.

Objective To analyze the effects of adenohypophysis function after treating with thyrotropin releasing hormone and its clinical significance in patients with brain trauma. Methods There were 22 cases with traumatic brain injuries from July 2010 to September 2012 in Chinese people's Liberation Army nine eight hospital,after injuried within 4 to 12 hours,then 1 1 cases who were given thyrotropin-releasing hormone(TRH)were selected as experimental group,while 1 1 cases who were given the same amount of isotonic saline were selected as control group,then the score of GCS, ICS,RLS85 and the improvement of adenohypophysis function were observed. Results After treatment,the score of glasgow coma scale (GCS ), innsbruck coma scale (ICS),and the reaction level scale (RLS85)between two groups were significantly increased in three days compared with before treatment,and within three days after injury situation,the improvements of ICS and RLS85 in experimental group were better than control group(P<0.05 ). Compared with control group,the levels of each gland pituitary hormone in experimental group were significantly increased(P<0.05 ),and on the third day,the growth hormone (GH)was reduced significantly,finally 50%of that in control group. Conclusion Patients with brain injury treated with thyrotropin releasing hormone,has no significant adverse reactions,with the characteristics of safe and effective.

Objective To investigate the clinical significance of serum anti-Saccharomyces cerevisias antibody (ASCA),anti-outer membrane porin C (anti-OmpC),antibody to Pseudomonas fluorescens-associated sequence I2 (anti-I2 )and antibody to bacterial flagellin (anti-CBirl )in the diagnosis and treatment of inflammatory bowel disease (IBD).Methods From 2011 to 2013,87 patients with IBD were enrolled and divided into Crohn′s disease (CD)group (66 cases)and ulcerative colitis (UC)group (21 cases).A total of 62 age and gender matched healthy individuals were enrolled as the control group. Fasting blood samples (2 mL)of the subjects were collected.The expression of ASCA,anti-OmpC,anti-I2 and anti-Cbirl antibodies was detected with enzyme-linked immunosorbent assay (ELISA)kits.The diagnosis value of each antibody in IBD and the differential diagnostic value of in UC and CD were compared by receiver operating characteristic (ROC)curve.Results The area under the curve (AUC)of ASCA between IBD and the healthy control group,between CD group and UC group was 0.580 and 0.512, respectively;the accuracy in diagnosis was low.The AUC of anti-CBirl between IBD and the healthy control group was 0.617.There was no differential diagnosis significance of the other antibodies.The positive rate of ASCA in IBD group was 62.1 % (54/87),which was significantly higher than that in the control group (38.7%,24/62).The positive rates of anti-OmpC and anti-I2 in IBD group was significantly lower than those in the control group and the differences were statistically significant (both P <0.05).No difference was observed in positive rates of serum antibodies among the others groups (all P >0.05).The specificity,sensitivity,positive predictive value (PPV)and negative predictive value (NPV)of ASCA in differential diagnosis of CD and UC was 52.4%,66.7%,81 .48% and 33.33%,respectively.The specificity and sensitivity of anti-OmpC,anti-I2 and anti-CBirl in differential diagnosis of CD and UC was 81 .0% to 100.0% and 9.1 % to 37.9%,respectively.The specificity,sensitivity,PPV and NPV of double-positive ASCA and anti-I2 in the diagnosis of CD was 57.1 %,86.4%,82.6% and 50.0%, respectively.The positive rate of ASCA and anti-I2 in CD group was significantly higher than that in UC group (84.8%(56/66)vs 57.1 % (12/21 );χ2 =5 .633,P =0.018 ).Conclusions Positive ASCA has some significance in the diagnosis of patients with IBD in our country.The detection of anti-I2 can help to diagnose ASCA negative CD.Because of low sensitivity and positive rate,anti-OmpC and anti-CBirl have limited value in the diagnosis of IBD and the differential diagnosis of UC and CD in our country.

Objective To study the characteristics of skinfold thickness of Han adults in Jiangsu province . Methods The skinfold thicknesses of facial , subscapular , suprailiac , biceps , triceps and calf on 311 urban adults ( 157 males and 154 females) and 421 rural adults ( 213 males and 208 females ) of Han were investigated in Huaian city of Jiangsu province .Results The thickness of skinfold of urban females were thicker than that of urban males .Rural adults were the same .Han adults of Jiangsu showed the most significant differences between urban areas and rural areas .The values of six skinfold thicknesses of Jiangsu urban adults have positive correlation with age .Conclusion Han adults of Jiangsu show the most significant differences between genders .

The American Clinical Practice Guiding -Neurogenic Bowel Management in Adults with Spinal Cord Injury (SCI) had been prepared based on scientific and professional information known about neurogenic bowel management, its causes, and its treatment, in 1998. It will provide reference to China's neurogenic bowel management after SCI in rehabilitation and treatment.