Abstract: Objective　To understand the characteristics of drug resistance and molecular transmission network of HIV/AIDS patients aged 50 years and above in Zhaotong City, Yunnan Province, from 2020 to 2022, who have failed antiretroviral therapy (plasma viral load≥1 000 copies/mL). Methods　The demographic data of HIV/AIDS patients aged 50 years and above with failed antiviral therapy in Zhaotong City from 2020 to 2022 were collected through the antiviral therapy database of Yunnan Province. The plasma of the patients was collected, and the HIV pol gene protease and reverse transcriptase regions were amplified by reverse transcriptase chain reaction, the fragments were spliced using ContigExpress, the sequences were determined, and drug resistance mutations were analyzed using the HIVdb database of Stanford University. Pairwise genetic distances between sequences were calculated using the HyPhy 2.2.4 software with the TN93 model, and a threshold of 1.7% was used to construct the HIV molecular network. Cytoscape 3.7.0 was used to visualize the network. Results　A total of 480 HIV/AIDS cases were included, with 417 positive amplifications of plasma samples. Among these, 196 cases were drug-resistant, resulting in a total drug resistance rate of 47.0%. The highest resistance rate was observed in NNRTIs at 45.6% (190/417), followed by NRTIs at 14.4% (60/417), and PIs at a lower rate of 0.2% (1/417). In NNRTIs resistance mutation sites, K103N/E/S mutation sites were dominant, with a mutation rate of 33.3% (139/417). Among the NRTIs resistance mutation sites, M184V/I mutation site was the main one, with a mutation rate of 13.9% (58/417). The mutation rate of PIs resistance sites was relatively lower, at 0.2% (1/417). Through the construction of the molecular transmission network, 88 patients entered the molecular transmission network, with an access rate of 21.1%, forming 21 transmission clusters, among which CRF01_AE formed a large cluster including 37 individuals. Conclusions　Compared with studies in other regions of Yunnan Province, the drug resistance rate among the elderly population with antiretroviral therapy failure in this study in Zhaotong City has decreased, and URFs have become the main subtype, suggesting that the transmission relationships among the elderly in Zhaotong City are gradually complicated. Through the construction of a molecular network, it is suggested that CRF01_AE and married or partnered males have the highest network access rate. Interventions should be strengthened among key populations, and diversified efforts in HIV/AIDS prevention and treatment for the elderly should be developed.

To study the correlation between traditional Chinese medicine (TCM) syndrome type and the ultrasound imaging changes in patients with limb lymphedema, and to provide evidence for TCM syndrome differentiation.

AimTo investigate the immune effects of the genetic engineering vaccine against swine cysticercosis with the animal mode of kunming mouse. Method78 kunming mice were randomly divided into three groups: 1 )injection one times with the vaccine, 2)injection twice with the vaccine at intervals of ten days, 3)non-vaccination group. All the mice were infected with the vial hatched oncopheres by the tag vein at 20 days after the first vaccination,and were killed and dissected to look for swine cysticerci 63 days later. In order to know the immune response of the vaccine and the stateof parasites in the rats,the antigens and antibodies of swine cysticerci were detected by immune methods. ResultsThe vaccine was safety to all the vaccinated rats. The antibodies against swine cysticerci began to appear positive in some of the immunized mice at 7 days postvaccination,and they were all positive at 21 days. 65 cysticerci were found in the mice of the nonvaccination group,which lodged mainly in the chests. 2 and lcysticerci were respectively found in two rats of the first group and in one rat of the second group, their protective rates of cysticerci were 96.9％ and 98. 4％,respectively. ConclusionKunming mouse as the animal model of swine cysticercosis was stable and reliable,and the genetic engineering vaccine against swine cysticercosis was safety and efficiency.

Objective To establish an efficient HPLC method for the determination of uric acid in plasma of hyperuricemia model mice,and the evaluation of uric acid lowering effect of Lesinurad.Methods The Laballiance Series Ⅲ HPLC system was adopted with Kromasil C18 column (100 mm × 4.6 mm,5 μm).The mobile phase consisted of methanol-0.5％ acetic acid (10:90) for isocratic elution with a flow rate of 0.4 mL/min.The detection wavelength was set at 283 nm.The established HPLC method was used to detect the plasma uric acid level of mice at 0.5,1.0,and 2.0 h time points after which being ip injected with 250 and 500 mg/kg uric acid.Lesinurad of 250 and 500 mg/kg was ig given to mice,0.5 h later,mice were ip injected with 500 mg/kg uric acid to establish hyperuricemia model,and 1 h later,the established HPLC method was used to detect the plasma uric acid level of mice.Results There was a good linear relationship between peak area and the concentration of plasma uric acid in the range of 7.5-150 μg/mL (r =0.997).The specificity,repeatability,precision,stability,and recovery of the established HPLC method was in accordance with the guiding rules of biological sample determination.Compared with the endogenous serum uric acid concentration of control group mice,serum uric acid concentration of 250 mg/kg dose group was significantly increased 0.5 h after ip administration with uric acid (P ＜ 0.01),and serum uric acid concentration of 500 mg/kg dose group was significantly increased 0.5,1.0,and 2.0 h after ip administration with uric acid.Compared with model group,the concentration of uric acid in plasma decreased significantly in low dosage group administered with Lesinurad (P ＜ 0.05),while decreased more significantly in high dosage group (P ＜ 0.01).Conclusion This convenient,rapid,and accurate method can be applied to the determination of uric acid in mouse plasma and the evaluation of relative drugs,which provide an efficient analysis way for establishing hyperuricemia model and screening relative drugs.

Objective: To investigate the causal relationship between gut microbiota and polycystic ovary syndrome (PCOS) using a Mendelian randomization (MR) study, so as to provide insights into the pathogenesis of PCOS and the formulation of prevention and treatment strategies. Methods: The genetic data on gut microbiota was derived from a meta-analysis of genome-wide association studies (GWAS) involving 18 340 participants. The genetic data on PCOS was sourced from two GWAS meta-analyses in European populations, serving as the discovery set and the validation set, respectively. A two-sample MR analysis was conducted using the discovery set, with the inverse variance weighted (IVW) method as the primary approach. Sensitivity analyses employed the weighted median method, MR-Egger regression, and the MR-PRESSO test. The validation set was utilized for verification, and a meta-analysis was performed to combine the results from the two datasets. Results: Forward MR analysis results showed that nine types of gut microbiota were statistically associated with PCOS (all P<0.05). Specifically, the association of family Streptococcaceae (OR=1.442, 95%CI: 1.097-1.895), genus Actinomyces (OR=1.359, 95%CI: 1.036-1.784), genus Ruminococcaceae UCG 011 (OR=0.755, 95%CI: 0.619-0.921), genus Sellimonas (OR=0.766, 95%CI: 0.657-0.893) and genus Streptococcus with PCOS (OR=1.496, 95%CI: 1.136-1.972) remained consistent in the sensitivity analysis. Reverse MR analysis showed no evidence for the causal association between PCOS and the aforementioned five types of gut microbiota (all P>0.05). The MR analysis results of the validation set showed that there was no statistical association between the aforementioned five types of gut microbiota and PCOS (all P>0.05). However, the associations remained significant for genus Actinomyces (OR=1.226,95%CI：1.010-1.503) and genus Streptococcus (OR=1.266,95%CI：1.042-1.452) in the meta-analysis (both P<0.05). Conclusion This study provides the evidence that genus Actinomyces and genus Streptococcus are causally associated with PCOS.

AIM: To construct the prokaryotic expression system containing protein transduction domain (PTD) with heat shock protein 27 (HSP27) in order to prepare and purify the recombinant protein , and to verify whether the recombinant protein PTD-HSP27 has the ability to penetrate the human lens epithelial cell ( HLEC) membrane and the rabbit cornea.METHODS:The plasmid pKYB-PTD-HSPB1-6His was constructed by the technique of overlap extension PCR.The plasmid was transformed and PTD-HSP27 was purified through nickel affinity chromatography column and identi-fied by Western blotting.PTD-HSP27-6His was labeled with the fluorescein isothiocyanate (FITC).The penetrating ability of PTD-HSP27 into HLECs and rabbit cornea was tested .RESULTS:The recombinant PTD-HSP27 plasmid was success-fully cloned and effectively expressed .The correctness of the recombinant protein PTD-HSP27 was demonstrated .Fluores-cence microscopic examination showed that PTD-HSP27-FITC was internalized by HLECs .Fluorescent labeled PTD-HSP27 was then observed in the rabbit aqueous humor .CONCLUSION:The recombined gene PTD-HSPB1 was constructed by o-verlap extension PCR technique and the PTD-HSP27 fusion protein was prepared and purified by nickel affinity chromatog-raphy column.Using the technique of PTD-fusion protein, HSP27 was transduced into HLECs and passed through the cor-nea .

Objective To investigate the effect of surgical treatment of adult mandibular condylar fractures (including intracapsular fracture). Methods Thirty-two patients (33 sides) with condylar fractures underwent open reduction and rigid fixation. Six patients with intracapsular condylar fractures were treated with two 18-mm positional screws through a preauricular approach. Six patients (7 sides) with condylar neck fractures were rigidly fixed by 1 mini-plate via a retromandibular transparotid approach. Twenty patients with subcondylar fractures were operated and fixed by two titanium plates using an angular approach. Results The mean follow-up period was 13.5 months, and the mean maximum mouth opening was 37.5 mm by the last visit. All patients acquired good occlusal relationship and mandibular symmetry. Seven patients (21.9%) experienced transient palsy of the branches of the facial nerve, and recovered completely after three months. One patient developed a salivary fistula, and healed after two weeks of gauze compression. No permanent deficit of any facial nerve branch was observed. No patient showed condylar head resorption. Conclusion Appropriate surgical approaches and fixation methods for different types of condylar fractures are the key factors to achieve reliable clinical results.

Objective To evaluate the relationship between echocardiographic epicardial adipose tissue thickness(EAT) and the presence and severity of coronary artery disease(CAD). Methods One hundredand forty-seven patients (101 patients with CAD and 46 patients with normal coronary arteries by diagnostic coronary angiography) were enrolled. EAT thickness was measured using 2-D echocardiographic parasternal long-and short-axis views. EAT thickness measurements were compared with angiographic findings. Results EAT was significantly higher in CAD group comparison to control group [(7.41 ± 1.63)mm vs (4.41±1.60) mm, P ＜0.01 ]. Furthermore, EAT increased with the severity of CAD [(8.53 ± 1.00)mm vs (6.36 ±1.73)mm, P ＜0.01]. Gensini's score significantly correlated with EAT (r = 0.71, P ＜0.01 ). EAT thickness ≥5.35 mm had 87.13% sensitivity and 80.42% specificity (ROC area 0. 89, P = 0.01,95% CI [0.84 - 0.9;]) for predicting CAD. Conclusions EAT thickness, which is easily and non-invasively evaluated by transthoracic echocardiography, can be an adjunctive marker to classical risk factors for the prediction of CAD, it was significantly correlated with the severity of coronary artery disease.

Objective To retrospectively analyse pathogenic bacteria isolated from inpatients with lupus erythematosus (SLE) and lupus nephritis (SLE‐LN ) ,and provide references for diagnosis and treatment for these patients with infection . Methods A total of 380 inpatients diagnosed with SLE/SLE‐LN in our hospital from 2010 to 2014 were enrolled in this study ,in‐cluding 96 cases of patients with SLE‐LN .Bacterial inoculation ,culture ,isolation ,identification and drug sensitivity test were carried out .Statistical analysis and susceptibility analysis was performed by using the SPSS 19 .0 and WHONET5 .6 software .Results For patients with SLE and SLE‐LN ,urinary tract infection accounted for 25 .0% and 27 .1% ,hematogenous infection accounted for 8 .1% and 10 .4% ,skin tissue infection accounted for 12 .0% and 8 .3% ,respectively .The most common gram negative bacteria was Escherichia coli ,which accounted for 25 .53% and 30 .21% in patients with SLE and patients with SLE‐LN ,respectively .Followed by Bauman Acinetobacter ,which accounted for 13 .42% and 14 .54% in patients with SLE and patients with SLE‐LN ,respectively . The most common gram positive bacteria was Staphylococcus aureus ,which accounted for 11 .58% and 11 .46% in patients with SLE and patients with SLE‐LN ,respectively .Strains of Escherichia coli were isolated from urine specimens of 69 .79% of patients with SLE and 66 .67% patients with SLE‐LN ,the percentages were significantly higher than that of the conventional urine culture (45% ,P< 0 .01) .The resistance rate of Escherichia coli strains isolated from patients with SLE to quinolones was higher than 66 .00% ,the resistance rate to ampicillin was 89 .69% ,and the resistance rate to piperacillin/tazobactam was low (3 .09% ) .The iso‐lation rates of ESBLs‐producing Escherichia coli strains and ESBLs‐producing Klebsiella pneumoniae strains in patients with SLE‐LN were higher than those in patients with SLE .Conclusion The patients with SLE have a higher risk for infection .The beta‐lac‐tams could be used for the treatment of Escherichia coli urinary tract infection in patients with SLE .

Objective: To evaluate the association of smoking with the risk of ankylosing spondylitis (AS) using a Mendelian randomization (MR) approach. Methods: A total of 16 383 186 AS-associated single nucleotide polymorphisms (SNPs), 378 smoking initiation associated SNPs and 126 lifetime smoking score-associated SNPs were collected from three large-scale genome-wide association studies (GWAS). The association of smoking phenotypes with the risk of AS was examined using inverse-variance weighted (IVW) with AS as a outcome variable, smoking initiation and lifetime smoking score as exposure factors and SNPs with strong associations with smoking as instrumental variables, and sensitivity analyses were performed with maximum likelihood-based method, MR pleiotropy residual sum and outlier (MR-PRESSO) test and MR-Egger regression analysis. Results: A 33.5% increased risk of AS was found among genetically predicted smokers relative to non-smokers (OR=1.335, 95%CI: 1.059-1.682), and an increase in predicted lifetime smoking by per standard deviation resulted in a 101.4% increased risk of AS (OR=2.014, 95%CI: 1.341-3.024). The maximum likelihood-based method and MR-PRESSO test showed consistent correlated effect estimations and MR-Egger regression analysis identified no evidence of pleiotropy. Conclusion It is genetically predicted that smoking is associated with an increased risk of AS.