Abstract: Objective To analyze the resistance and spatial distribution of Mycobacterium tuberculosis (MTB) to six commonly used anti-tuberculosis drugs in Qinghai Province from 2016 to 2019, so as to provide a reference for tuberculosis treatment and drug-resistant tuberculosis control. Methods A total of 1 182 identified strains of Mycobacterium tuberculosis in Qinghai Province from 2016 to 2019 were collected, and 6 anti-tuberculosis drugs were subjected to drug susceptibility tests and strain confirmed by the proportional method. By means of ArcMap10.7 and SaTScan10.1 software, map visualization, spatial autocorrelation analysis and spatial scanning of MTB drug resistance were performed to identify MTB drug resistance clusters in Qinghai Province. Results From 2016 to 2019, the total drug resistance (TDR) rate of 1 182 Mycobacterium tuberculosis strains in Qinghai Province was 23.77% (281/1 182), with a mono-resistance (MR) rate of 11.08% (131/1 182), a poly-resistance (PDR) rate of 3.89% (46/1 182), a multi-drug resistance (MDR) rate of 8.80% (104/1 182), and an extensive drug resistance (XDR) rate of 0.85% (10/1 182). The rates of MDR, XDR and TDR all showed a decreasing trend year by year (P<0.01). The drug resistance spectrum displayed 21 combinations. The TDR rate and MDR rate in the retreatment patients were higher than those of the initial treated patients, and the difference was statistically significant (χ2 TDR=22.784, χ2MDR=45.082, P<0.01). In terms of demographic characteristics, the TDR rate in males was higher than that in females, and the middle-aged group was higher than other age groups, and the differences were statistically significant (χ2=7.541, 10.825, P<0.05). The results of global spatial autocorrelation analysis showed that there was no statistical significance in the autocorrelation and obvious spatial clustering of MTB drug resistance in Qinghai Province from 2016 to 2019 (P>0.05), which indicated a random distribution. The results of spatiotemporal scanning showed that there was a kind of clustering area, but the clustering effect was not significant (P>0.05), indicating a random distribution. Conclusions The TDR of MTB in Qinghai Province from 2016 to 2019 showed a downward trend year by year. In comparison with the national average, the rate of multi-drug resistance and extensive drug resistance was still high, and most of the multi-drug resistance resulted from rifampicin and isoniazid. The drugresistant population mainly consisted of retreatment, males, and young and middle-aged pop

OBJECTIVETo explore the correlation between F10 gene mutation and its phenotype in a Chinese pedigree affected with FX deficiency.

METHODSProthrombin time(PT), activated partial thromboplastin time(APTT), fibrinogen, FII activity(FII:C), FVII activity(FVII:C), FIX activity (FIX:C), FX activity(FX:C) were determined with a one-stage clotting assay. The FX antigen(FX:Ag) was detected with an enzyme linked immunosorbent assay(ELISA). The 8 exons, introns and 5' and 3' untranslated regions(UTR) of the F10 gene of the proband and her family members were subjected to PCR amplification and Sanger sequencing. Suspected mutation was confirmed by reverse sequencing. Polymorphisms were excluded by direct sequencing of 100 healthy individuals.

RESULTSThe PT and APTT of the proband have prolonged to 16.1 s and 49.0 s, respectively. Her FX:C and FX:Ag were reduced by 27% and 56%, and her mother's PT, APTT, FX:C and FX:Ag were 14.8 s, 37.4 s, 44%, 34%, respectively. Her grandmother's PT, APTT, FX:C and FX:Ag were 15.8 s, 42.2 s, 31%, 45%, respectively. The results of her father and other family members were all within the normal range. Genetic analysis has revealed a heterozygous G to A mutation in the proband at position 28076 in exon 8 of the F10 gene, which resulted in a p.Gly363Ser substitution. The same mutation was also found in her mother and grandmother. No mutation of the F10 gene was found in her father. Gly363Ser may result in changes in the secondary structure of the FX protein and reduction of its activity.

CONCLUSIONThe g.28076G to A(p.Gly363Ser) mutation of the F10 gene probably underlies the FX deficiency in this pedigree. The mutation was discovered for the first time in Chinese patients.

OBJECTIVETo investigate the phenotype and genotype defect characteristics of a Chinese patient with hereditary factor XI deficiency.

METHODSThe activated partial thromboplastin time (APTT), prothrombin time (PT), FXI activity (FXI:C) of the proband and his relatives were measured by a clotting method using automatic coagulation analyzer. FXI antigen (FXI:Ag) was assayed by enzyme-linked immunosorbent assay (ELISA). Fifteen exons of the F11 gene were amplified by PCR and sequenced. Pymol software was used to analyze the novel mutations.

RESULTSThe APTT of the proband was significantly prolonged (70.3 s, reference 34.5 s) with decreased FXI activity (6%, reference 50%-150%) and FXI antigen (1.9%, reference 50%-150%). The FXI activity and FXI antigen of his son was 31% and 39%, respectively. Two heterozygous F11 mutations were identified in the proband, which included a G to T substitution at nucleotide 1296 in exon 11 resulting in substitution of glycine by valine at codon 400 (p.Gly400Val) and a A to T substitution at nucleotide 1691 in exon 14 resulting in substitution of arginine (AGA) by a termination codon (TGA) at codon 532 (p.Arg532Ter). Analysis using Pymol indicated that the number of hydrogen bonds has changed, which led to a transformation of the structure of the FXI protein. The son of the proband was found to be heterozygous for the c.1296G to T (p.Gly400Val) mutation. NM_13142 c.1691A to T (p.Arg532Ter) is a novel mutation based on HGMD professional 2016.4. Based on 2015 Guidelines of ACMG, it is PVS1 (very strong pathogenicity).

CONCLUSIONThe compound heterozygous mutations of F11 NM_13142 c.1296G to T (p.Gly400Val) and F11 NM_13142 c.1691A to T(p.Arg532Ter) probably underlies the FXI deficiency in the proband.

Objective: The study aims to explore the epidemiological characteristics of tuberculosis among students in Qinghai Province, to provide scientific basis for the prevention and control of students tuberculosis. Methods: Data on tuberculosis among students from 2016 to 2019 in Qinghai province were collected and epidemiological characteristics were analyzed, the spatial distribution map were drawn by using ArcMap 10.8. Results: During 2016-2019, there were 2 691 reported cases of tuberculosis among students in Qinghai Province the reporting rate were 46.10/10 5, 68.50/10 5, 73.49/10 5, 85.96/10 5, increased year by year( χ 2=116.45, P <0.01). With a high incidence from March to September each year. The tuberculosis patients were mainly aged 18 years and above, with more reported female cases than male cases and more Tibetan cases. Most of students tuberculosis cases were reported in southern Qinghai, especially in Yushu and Guoluo areas, and sharp increase was observed in Xining during 2018 to 2019. Conclusion Students tuberculosis in Qinghai is still serious. Schools should strengthen education on tuberculosis prevention, especially those in southern Qinghai and Xining.

OBJECTIVE: To explore molecular etiology and clinical characteristics of two pedigrees affected with hereditary factor VII(FVII) deficiency. METHODS: The nine exons and flanking sequences of the F7 gene of the probands were amplified by PCR. The amplicons were analyzed by direct sequencing. Suspected mutations were subjected to SWISS-MODEL modeling and analysis of protein structure change by Pymol software and conservation of amino acids across various species. RESULTS: For proband of pedigree 1, the prothrombin time (PT), FVII activity (FVII:C) and FVII antigen (FVII:Ag) were 36.3 s, 3%, 53.56%, respectively. Sequencing revealed a compound heterozygous variants of c.80_81delCT and c.1371G>T(p.Arg439Ser). His son carried a heterozygous c.1371G>T (p.Arg439Ser) variant. For proband of pedigree 2, the PT, FVII:C and FVII:Ag were 22.3 s, 4%, 1.58%, respectively. Sequencing has revealed a compound heterozygous c.278G>T(p.Arg75Met) missense variant in exon 3 and c.1278T>G (p.His408Gln) in exon 9 of the F7 gene. His mother and son both carried a heterozygous c.278G>T(p.Arg75Met) variant. Three-dimensional simulation and homology analysis revealed that the p.Arg439Ser and p.Arg75Met can respectively alter part of hydrogen bonds and two highly conserved amino acids. CONCLUSION Two novel heterozygous missense variants of the F7 gene [c.1371G>T(p.Arg439Ser) and c.278G>T(p.Arg75Met)] probably account for the decrease of factor VII in the two pedigrees.
Asian Continental Ancestry Group ; Factor VII ; Factor VII Deficiency ; Genotype ; Heterozygote ; Humans ; Mutation ; Pedigree

OBJECTIVE: To carry out phenotypic and genotypic analysis for two Chinese pedigrees affected with coagulation factor XII (F XII) deficiency. METHODS: Plasma prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), thrombin time (TT), and blood coagulation factor VIII, IX, XI, XII activity (FVIII:C, FIX:C, FXI:C, FXII:C) were determined with one stage clotting assay on a STAGO coagulation analyzer. FXII antigen was determined with an enzyme linked immunosorbent assay (ELISA). The 14 exons and their flanking sequences of the F12 gene were subjected to PCR amplification and Sanger sequencing. The conservation and structure of mutant protein were analyzed with MegAlign software and PYMOL software. RESULTS: The APTT of the probands was significantly prolonged, while their FXII:C and FXII:Ag were significantly reduced. Genetic analysis of the proband has revealed three novel mutations in the F12 gene, including g.5972G>A splice site mutation in intron 5, g.8810_8814delGTCTA in exon 14, and g.6259G>A (p.Pro182Leu) in exon 7. In addition, a previously known mutation IVS13-1G>A has been found. CONCLUSION Four mutations have been identified in the two Chinese pedigrees, among which three were novel. Above mutations probably played a role in the defect of FXII in the two pedigrees.
Exons ; Factor XII ; genetics ; Factor XII Deficiency ; genetics ; Genetic Testing ; Humans ; Pedigree

Objective: To analyze the phenotype and genotype of a pedigree affected with congenital dysfibrinogenemia. Methods: Liver and kidney functions of the proband and her relatives were determined. Coagulation tests including prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time(TT), fibrin(ogen) degradation products (FDPs), D-dimer(D-D) and the calibration experiment of protamine sulfate of against plasma TT were detected in the proband and her predigree members. The activity and antigen of fibrinogen (Fg) in plasma were measured by Clauss method and immunonephelometry method, respectively. All of the exons and exons-intron boundaries of the three fibrinogen genes (FGA, FGB and FGG) were subjected to PCR amplification and Sanger sequencing. Potential influence of the suspected mutations were analyzed with bioinformatics software including PolyPhen-2, SIFT and Mutation Taster. Results: The proband had normal PT, APTT, FDPs, D-D and prolonged TT (31.8 s). The activity of fibrinogen (Fg) in plasma was significantly decreased but the antigen was normal. Genetic analysis revealed a heterozygous c. 92G>A (p.Gly31Glu) mutation in exon 2 of the FGA gene. Family studies revealed that the mother carried the same mutation. Bioinformatic analysis suggested that the mutation may affect the function of Fg Protein. Conclusion The dysfibrinogenemia was probably caused by the novel Gly31Glu mutation of the FGA gene.

OBJECTIVE: To analyze the phenotype and genotype of a pedigree affected with congenital dysfibrinogenemia. METHODS: Liver and kidney functions of the proband and her relatives were determined. Coagulation tests including prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time(TT), fibrin(ogen) degradation products (FDPs), D-dimer(D-D) and the calibration experiment of protamine sulfate of against plasma TT were detected in the proband and her predigree members. The activity and antigen of fibrinogen (Fg) in plasma were measured by Clauss method and immunonephelometry method, respectively. All of the exons and exons-intron boundaries of the three fibrinogen genes (FGA, FGB and FGG) were subjected to PCR amplification and Sanger sequencing. Potential influence of the suspected mutations were analyzed with bioinformatics software including PolyPhen-2, SIFT and Mutation Taster. RESULTS: The proband had normal PT, APTT, FDPs, D-D and prolonged TT (31.8 s). The activity of fibrinogen (Fg) in plasma was significantly decreased but the antigen was normal. Genetic analysis revealed a heterozygous c.92G>A (p.Gly31Glu) mutation in exon 2 of the FGA gene. Family studies revealed that the mother carried the same mutation. Bioinformatic analysis suggested that the mutation may affect the function of Fg Protein. CONCLUSION The dysfibrinogenemia was probably caused by the novel Gly31Glu mutation of the FGA gene.
Afibrinogenemia ; congenital ; genetics ; DNA Mutational Analysis ; Female ; Fibrinogen ; genetics ; Humans ; Mutation ; Pedigree ; Phenotype

Objective To analyze the current situation, change trend and influencing factors of the delay in the treatment of tuberculosis patients in Xianning from 2008 to 2021, and to provide a scientific basis for controlling the tuberculosis epidemic. Methods The current situation and trend of the delay in the treatment of tuberculosis patients in Xianning City from 2008 to 2021 were described, and the multivariate Logistic regression analysis model was used to explore the influencing factors of the treatment delay in tuberculosis patients. Results The delay rate of seeking medical care for tuberculosis patients in Xianning from 2008 to 2021 was 58.53%, and the delay rate in the first six years tended to be stable. From 2014 to 2016, it increased slightly and then decreased year by year to 44.76% in 2021. Different gender and age groups showed the same trends as overall patients. The analysis results of the multivariate Logistic regression analysis model showed that females, household registration address in Jiayu County, occupations including service industries, workers, farmers and herdsmen, retirees, and others, the diagnosis result being positive for etiology, and the direct treatment in the patient source were high-risk factors for patients with delayed treatment. The risk of delay in treatment in ≥ 65-year-old patients was lower than that of < 25-year-old patients. Conclusion From 2008 to 2021, the phenomenon of delay in the treatment of patients with pulmonary tuberculosis in Xianning City is relatively common, and the high-risk factors of the delay in treatment need to be paid more attention to.