Objective To investigate the application of the multiple locus variable numbers of tandem repeats(MLVA) in genotype Mycobacterium tuberculosis(MTB) strains isolated from Uygur in south of Xinjiang, and to understand the characteristics of genotype and distribution. Methods One hundred and fifty-one Mycobacterium tuberculosis strains were collected from Uygur in south of Xinjiang which contains three regions, Kashgar, Hotan and Kizilsu kirghiz. Twenty-four tandem repeats loci in the total genome of MTB were analyzed by PCR and agarose gel electrophoresis method. The characteristics on polymorphism of DNA fingerprinting of 151 MTB strains were analyzed with BioNumerics 5.0 software. Results Twenty-four MLVA loci of 151 MTB strains were analyzed respectively. The results showed that there were obvious polymorphisms of VNTRs. The clustering of genotype showed that these strains could be categorized into 8 gene groups( Ⅰ , Ⅱ , Ⅲ ,Ⅳ, Ⅴ ,Ⅵ,Ⅶ ,Ⅷ) and 151 genotypes. Sixty-seven isolates(44.4% ) belonged to group Ⅵ. 23.2% were group Ⅷ including 35 genotypes, 20. 5% were group Ⅳ including 31 genotypes. The group Ⅵ prevailed mostly in the Kashgar. The group Ⅲ prevailed mostly in the HOTAN. Conclusion The results showed there were obvious polymorphisms of VNTRs of Mycobacterium tuberculosis clinical strains preliminarily. Group Ⅵ was the predominant prevalent strain in south of Xinjiang.

Objective To construct lentivirus containing CXCR4 gene and transfect MCF-7 cells , and obtain CXCR4 high-expressing MCF-7 cells. Methods CXCR4 gene was amplified by RT-PCR to construct CXCR4/pSin-EF2, which was transfected into HEK293T cells with psPAX2 and pMD2G vector for lentivirus packing. Packaged lentivirus was used to transfect human breast cancer cells MCF-7, with empty lentivirus as control. CXCR4 mRNA and protein expression levels were detected by RT-PCR and Western blot before and after transfection. And flow cytometry was used to detecte cell surface CXCR4 expression. Results The recombinant plasmid CXCR4/pSin-EF2 was constructed successfully,identified by double digestion and sequencing, and transfected into HEK293T cells to obtain high-titer lentivirus. RT-PCR and Western blot confirmed that the expression of CXCR4 in MCF-7 cells increased significantly after CXCR4 lentivirus transfection. Flow cytometry results showed that the CXCR4 positive rate increased from 26.78% to 99.29%, while there is no significant difference in CXCR4 expression between vector-transfected MCF-7 cells and non-transfected MCF-7 cells. Conclusion CXCR4 lentivirus and the breast cancer cell line with high and stable expression of CXCR4 (MCF-7CXCR4) were successfully constructed.

Objective To study the awareness of perimenopausal hormone replacement therapy (HRT) among a part of the medical care personnel in Guiyang . Methods A survey was conducted among 500 medical staff members in 4 hospitals of Guiyang by cluster random sampling using questionnaire about HRT. Results The survey showed that 74.6% (373/500) medical staff thought that the hormone replacement therapy was necessary to perimenopausal women; 96.7% (87/90) of obstetrics and gynecology doctors believed that it was necessary for perimenopausal women to use HRT,which was significantly higher than the doctors of other specialties 68.6% (166/242) and the nurses group 71.4% (120/168) (χ2=28.509, 23.537, P<0.01). Only 5.8%(29/500) of the medical personnel were willing to recommend HRT. In light of the attitude for recommending HRT, the obstetricians and gynecologists group was more significantly higher than the other specialties doctors group (χ2=86.781, P<0.01). Conclusion The knowledge of hormone replacement therapy in part of Guiyang medical personnel is not sufficient;the recommending rate of HRT was low;the side effects of HRT was still a concern. There are differences between obstetrics and gynecology doctors and doctors other specialties and nurses in HRT knowledge.

OBJECTIVETo construct a soluble prokaryotic expression vector of the CXCR7-specific antagonist SDF-1/54R and evaluate its activity.

METHODSSDF-1/54r gene amplified by PCR was inserted into the soluble expression vector pET-41a+ engineered with GST fusion tag, and the recombinant vector was transformed into E. coli strain BL21 (DE3). After IPTG induction of E. coli, the expressed recombinant protein was purified with GST affinity chromatography purification system and confirmed by SDS-PAGE and Western blotting assay. The target protein SDF-1/54R was obtained after digestion of the purified product with enterokinase. Breast cancer MCF-7 cells with high expression of CXCR7 was treated with SDF-1/54R and the cell proliferation and metastasis was evaluated with MTT and chemotaxis assays.

RESULTSThe target protein SDF-1/54R obtained showed an obvious inhibitory effect on the proliferation and metastasis of MCF-7 cells as confirmed by MTT and chemotaxis assays.

CONCLUSIONSDF-1/54R is a good antagonist of CXCR7 and shows a potential value as an effective anti-cancer agent.

Blotting, Western ; Chemokine CXCL12 ; metabolism ; Chromatography, Affinity ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Genetic Vectors ; Humans ; Polymerase Chain Reaction ; Receptors, CXCR ; antagonists & inhibitors ; Recombinant Proteins ; biosynthesis

Iron is involved in the virulence and pathogenic effects of certain intracellular parasites.In the pathogenic process of Brucella,the uptaking and metabolism of host iron are closely related to intracellular parasitism and immunity escape of Brucella.In this paper,we elucidated the iron transport system,iron response regulators and nutrient immunity of iron based on the latest report and data about Brucella.

Objective To construct a soluble prokaryotic expression vector of the CXCR7-specific antagonist SDF-1/54R and evaluate its activity. Methods SDF-1/54r gene amplified by PCR was inserted into the soluble expression vector pET-41a+engineered with GST fusion tag, and the recombinant vector was transformed into E. coli strain BL21 (DE3). After IPTG induction of E. coli, the expressed recombinant protein was purified with GST affinity chromatography purification system and confirmed by SDS-PAGE and Western blotting assay. The target protein SDF-1/54R was obtained after digestion of the purified product with enterokinase. Breast cancer MCF-7 cells with high expression of CXCR7 was treated with SDF-1/54R and the cell proliferation and metastasis was evaluated with MTT and chemotaxis assays. Results The target protein SDF-1/54R obtained showed an obvious inhibitory effect on the proliferation and metastasis of MCF-7 cells as confirmed by MTT and chemotaxis assays. Conclusion SDF-1/54R is a good antagonist of CXCR7 and shows a potential value as an effective anti-cancer agent.

Objective To construct a soluble prokaryotic expression vector of the CXCR7-specific antagonist SDF-1/54R and evaluate its activity. Methods SDF-1/54r gene amplified by PCR was inserted into the soluble expression vector pET-41a+engineered with GST fusion tag, and the recombinant vector was transformed into E. coli strain BL21 (DE3). After IPTG induction of E. coli, the expressed recombinant protein was purified with GST affinity chromatography purification system and confirmed by SDS-PAGE and Western blotting assay. The target protein SDF-1/54R was obtained after digestion of the purified product with enterokinase. Breast cancer MCF-7 cells with high expression of CXCR7 was treated with SDF-1/54R and the cell proliferation and metastasis was evaluated with MTT and chemotaxis assays. Results The target protein SDF-1/54R obtained showed an obvious inhibitory effect on the proliferation and metastasis of MCF-7 cells as confirmed by MTT and chemotaxis assays. Conclusion SDF-1/54R is a good antagonist of CXCR7 and shows a potential value as an effective anti-cancer agent.

Rheumatoid arthritis(RA)is a common systemic autoimmune disease.Chronic synovial inflammation,angiogene-sis,pannus formation,joint destruction of cartilage and bone are important pathological characteristics in RA.Semaphorins(Sema)is a kind of secreted and membrane-bound glycoprotein with nerve guiding effect.Recent studies have shown that Sema family members play an important role in the various pathological stages of RA.Therefore,this paper reviews and discusses the latest research progress on the involvement of Sema family members in different pathological links of RA,which can help to find new feasible targets for RA and provide new research ideas for the pathogenesis of RA.

In deep learning-based image registration, the deformable region with complex anatomical structures is an important factor affecting the accuracy of network registration. However, it is difficult for existing methods to pay attention to complex anatomical regions of images. At the same time, the receptive field of the convolutional neural network is limited by the size of its convolution kernel, and it is difficult to learn the relationship between the voxels with far spatial location, making it difficult to deal with the large region deformation problem. Aiming at the above two problems, this paper proposes a cascaded multi-level registration network model based on transformer, and equipped it with a difficult deformable region perceptron based on mean square error. The difficult deformation perceptron uses sliding window and floating window techniques to retrieve the registered images, obtain the difficult deformation coefficient of each voxel, and identify the regions with the worst registration effect. In this study, the cascaded multi-level registration network model adopts the difficult deformation perceptron for hierarchical connection, and the self-attention mechanism is used to extract global features in the basic registration network to optimize the registration results of different scales. The experimental results show that the method proposed in this paper can perform progressive registration of complex deformation regions, thereby optimizing the registration results of brain medical images, which has a good auxiliary effect on the clinical diagnosis of doctors.
Algorithms ; Neural Networks, Computer ; Image Processing, Computer-Assisted/methods*

Objective To investigate the liver disease phenotypes and clinical features of patients with different genotypes of Wilson's disease(WD).Methods A retrospective analysis was performed for 163 patients with WD who were diagnosed and underwent genetic testing in The Fifth Medical Center of Chinese PLA General Hospital from August 2008 to June 2023,and clinical manifestations,laboratory examination,pathological examination,imaging examination,and ATP7B genetic testing results were collected.According to ATP7B gene mutation,the patients were divided into groups as follows:R778L mutation group and non-R778L mutation group;P992L mutation group and non-P992L mutation group;truncation mutation group and non-truncation mutation group.Liver disease phenotypes and clinical features were analyzed for the patients with c.2333G>T/p.R778L mutation(R778L mutation),c.2975C>T/p.P992L mutation(P992L mutation),and truncation mutation of the ATP7B gene.The Mann-Whitney U test or the Kruskal-Wallis H test was used for comparison of continuous data between groups,and the chi-square test or the Fisher's exact test was used for comparison of categorical data between groups.Results The 163 patients with WD had varying severities of liver disease phenotypes,among whom 121(74.23%)were diagnosed with chronic liver disease,36(22.09%)were diagnosed with decompensated cirrhosis,and 6(3.68%)were diagnosed with fulminant WD,and in addition,there were 5 patients(2 with chronic liver disease and 3 with decompensated cirrhosis)with neurological abnormalities.For the 163 patients with WD,R778L mutation(with an allele frequency of 28.2%)was the most common mutation in the ATP7B gene,followed by P992L mutation(with an allele frequency of 12.6%),and truncation mutation showed an allele frequency of 11.0%.There was no significant difference in the distribution of the three mutations across different liver disease phenotypes(P>0.05).The R778L mutation group had a significantly lower level of ceruloplasmin(CP)than the non-R778L mutation group[0.04(0.02-0.08)g/L vs 0.08(0.03-0.13)g/L,Z=-2.889,P=0.004].Compared with the non-P992L mutation group,the P992L mutation group had significantly higher levels of alanine aminotransferase[135.0(80.5-237.0)U/L vs 80.5(36.0-173.3)U/L,Z=2.684,P=0.007]and aspartate aminotransferase[121.4(77.0-195.0)U/L vs 84.0(39.0-123.3)U/L,Z=3.388,P<0.001].Compared with the non-truncation mutation group,the truncation mutation group had significantly lower levels of CP[0.03(0.02-0.08)g/L vs 0.06(0.03-0.11)g/L,Z=-3.136,P=0.002]and serum copper[3.20(2.15-5.00)mg/L vs 4.20(2.60-7.50)mg/L,Z=-2.296,P=0.025].Conclusion R778L mutation,P992L mutation and truncation mutation are not associated with liver disease phenotype in WD patients;however,R778L mutation is associated with a lower level of CP,P992L mutation is associated with higher levels of ALT and AST,and truncation mutation is associated with lower levels of CP and serum copper.