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.

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 prepare multi-modality melanin-based lipid nanobubbles (MNBs) contrast agents,and to investigate their manifestations of ultrasound (US),photoacoustic (PA) and MRI in vitro.Methods MNBs and lipid nanobubbles (NBs) were prepared with the method of CHCl3-injection,freeze-drying and C3F8-inflation.The characteristics (shape,grain diameter,loading capacity of melanin and stability) of MNBs were obseved.US,PA and MRI were detected in vitro and the images were quantitatively analyzed.Results MNBs presented with homogenized size distribution,and transmission electron microscope (TEM) images demonstrated that the melanin particles were successfully entrapped into nanobubbles.Meanwhile,the loading capacity of melanin was 90.53 μg/mg.US signal increased in vitro with the rise of MNBs and NBs concentration.The ultrasonic manifestations of MNBs and NBs were the same,and the relative signal enhancement had no significant difference (P＞0.05).With the increased concentration of MNBs,the PA and MRI signals were stronger,but NBs showed no evident enhancement.Conclusion Multi-modality MNBs contrast agents are prepared successfully,which can enhance US,PA and MR imaging.

ObjectiveThis study aims to investigate the inhibitory effect of Wutoutang on pannus formation in adjuvant-induced arthritis （AIA） rats with wind-cold-dampness Bi syndrome and its potential mechanism. MethodA total of 40 male SD specific pathogen-free （SPF） rats were selected and divided into blank group， wind-cold-dampness Bi syndrome group ［Complete Freund's Adjuvant （CFA）， 200 μg］， Wutoutang group （15 g·kg^-1·d^-1）， and indometacin group （10 mg·kg^-1） according to random number table method. Except for the blank group， the other groups were given wind-cold-dampness stimulation before the CFA injection. After the rats were administered for 30 days， the basic conditions， onset time， arthritis index score， and foot swelling volume of AIA rats with wind-cold-dampness Bi syndrome were observed. Finally， peripheral arterial blood， ankle joint， and synovial tissue were taken. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum hypoxia-inducible factor-1α （HIF-1α）， vascular endothelial growth factor A （VEGFA） protein content， and rheumatism， including anti-O （ASO）， C-reactive protein （CRP）， and rheumatoid factor （RF）. Hematoxylin-eosin （HE） staining revealed the changes in joint histomorphology. Immunohistochemistry was used to detect the expression of HIF-1α and VEGFA， two important proteins in the ankle pathway. Quantitative real-time polymerase chain reaction （Real-time PCR） was used to reveal mRNA levels of HIF-1α， VEGFA， angiopoietin-1 （Ang-1）， and angiopoietin-2 （Ang-2） in rat synovial tissue. ResultThe foot swelling volume and arthritis score of AIA rats with wind-cold-dampness Bi syndrome were substantially higher （P<0.01） compared with the blank group. Serum CRP， RF， and ASO levels were considerably elevated （P<0.01）. HE staining showed obvious hyperplasia of ankle synovium and synovial inflammation， angiogenesis and pannus formation， and aggravated bone destruction， indicating successful modeling. After the intervention of Wutoutang， the onset time was delayed （P<0.01）. Foot swelling volume and arthritis score were decreased （P<0.01）. Serum CRP， RF， and ASO levels were significantly decreased （P<0.01）. The inflammatory hyperplasia of synovial tissue， angiogenesis and pannus formation， and bone destruction were alleviated. The mRNA levels of HIF-1α， VEGFA， Ang-1， and Ang-2 in the synovial membrane were significantly decreased （P<0.05， P<0.01）. The expressions of HIF-1α and VEGFA in serum and ankle joints were decreased （P<0.01）. In the indomethacin group， the onset time of the disease was delayed （P<0.01）. Foot swelling volume and arthritis score were decreased （P<0.01）. Serum CRP， RF， and ASO levels were significantly decreased （P<0.01）. HIF-1α/VEGFA/Ang signaling pathway was activated， and pathological tissue injury was improved. ConclusionWutoutang can delay the onset time of AIA rats with wind-cold-dampness Bi syndrome， reduce foot swelling volume， arthritis score， rheumatic activity， and improve joint histopathology. It can inhibit pannus formation， and its mechanism may be related to down-regulating the expression of the HIF-1α/VEGFA/Ang pathway.

ObjectiveTo investigate the liver disease phenotypes and clinical features of patients with different genotypes of Wilson’s disease （WD）. MethodsA 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. ResultsThe 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］. ConclusionR778L 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.