Objective To investigate the epidemiological characteristics of cross-county imported dengue fever cases within Yunnan province in 2023, so as to provide insights into formulation of preventive and control measures for intra-provincial spread of dengue fever. Methods All data pertaining cross-county imported dengue fever cases within Yunnan Province in 2023 were collected, and the temporal, regional and population distributions of the cases were descriptively analyzed. Results A total of 1 664 intra-provincial cross-county imported dengue fever cases were reported in 95 counties (cities, districts) cross 16 profectures (cities) in Yunnan Province in 2023, accounting for 12.34% of total cases in the province. Cross-county imported dengue fever cases were predominantly reported during the period between August and October (1 516 cases, 91.11% of total cases), and peaked in September (659 cases), with a single-day peak on October 8 (36 cases). During the period from September 4 to 10, five counties (cities) with local dengue fever epidemics, including Jinghong City of Xishuangbanna Dai Autonomous Prefecture, Gengma Dai and Wa Autonomous County of Lincang City, Ruili City of Dehong Dai and Jingpo Autonomous Prefecture, Mengla Coun ty of Xishuangbanna Dai Autonomous Prefecture, and Zhenkang County of Lincang City, exported 165 cross-county imported dengue fever cases to the rest of the province. Among the 1 644 intra-provincial cross-county imported dengue fever cases, the male to female ratio was 1.40∶1.00, and 1 329 cases were at ages of 15 to 55 years (79.87%), with farmers as the predominant occupation (886 cases, 53.25%). The top 5 counties (cities/districts) reporting the highest number of intra-provincial cross-county imported dengue fever cases included Simao District (266 cases) and Lancang Lahu Autonomous County (118 cases) of Pu’er City, Mengla County (91 cases) and Menghai County (91 cases) of Xishuangbanna Dai Autonomous Prefecture, and Mangshi City (73 cases) of Dehong Dai and Jingpo Autonomous Prefecture, which accounting for 38.40% of total imported cases. These intra-provincial cross-county imported dengue fever cases originated from 7 counties (cities/districts) in 4 prefectures (cities), including 1 261 cases (76.70%) from Jinghong City of Xishuangbanna Dai Autonomous Prefecture, 224 cases (13.63%) from Ruili City of Dehong Dai and Jingpo Autonomous Prefecture, 103 cases (6.27%) from Gengma Dai and Wa Autonomous County of Lincang City, 31 cases (1.89%) from Mengla County of Xishuangbanna Dai Autonomous Prefecture, 30 cases (1.82%) from Zhenkang County of Lincang City, 10 cases (0.61%) from Cangyuan Wa Autonomous County of Lincang City, and 5 cases (0.30%) from Mohan-Boten Economic Cooperation Zone of Kunming City. In addition, local dengue fever epidemics following intra-provincial cross-county importation of dengue fevers cases in Simao District, Jinggu Dai and Yi Autonomous County, Mangshi City, Longchuan County, and Cangyuan Wa Autonomous County. Conclusions Farmers and students are high-risk populations for intra-provincial cross-county imported dengue fever cases in Yunnan Province, and health education pertaining personal protection against dengue fever should be strengthened among these high-risk populations by governments at all levels. There is a high risk of local out-break of dengue fever following continuous introduction of intra-provincial cross-county imported cases. Standardized management of intra-provincial cross-county imported dengue fever cases should be reinforced to reduce the risk of local epidemics.

Objective:To explore the HIV-1 drug resistance in patients with HIV/AIDS in Guilin city following the failure of antiretroviral treatment (ART).Methods:Plasma samples were collected from patients in Guilin who had received ART for more than 1 year and had a HIV viral load greater than or equal to 1 000 copies/ml from January 2019 to December 2023, and demographic information was also collected for HIV-1 genotype subtype analysis and drug resistance testing to determine the resistance mutation loci and the susceptibility of the strains to drugs.Results:A total of 766 patient samples with failed ART collection and successful amplification were collected, of which 536 (69.97%, 536/766) were male, with an average age of 53 years; a total of 8 HIV-1 subtypes were detected, with CRF01_AE (80.55%, 617/766), CRF07_BC (11.10%, 85/766) and CRF08_BC (6.92%, 53/766) predominated. The drug resistance analysis showed that the HIV-1 drug resistance rate was 34.86% (267/766), including nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs) and protease inhibitor (PI), with dual resistance to NRTIs/NNRTIs (48.31%, 129/267) and NNRTIs resistance (43.07%, 115/267) predominantly. A total of 37 resistance mutation sites were detected, 14 NRTIs-associated mutation sites mainly included M184V/I (47.57%, 127/267), K65R (18.73%, 50/267), K70E/N/T/G/R (13.11%, 35/267), etc., and 18 NNRTIs-associated mutation sites mainly included K103 N/R (56.93%, 152/267), V179 D/E/T (21.72%, 58/267), G190C/S/Q (17.23%, 46/267), and V106I/M (16.85%, 45/267), etc.; and 5 PIs-associated mutation sites was the highest with L10V/I mutation rate (3.00%, 8/267).Conclusions:HIV/AIDS patients in Guilin have shown favorable outcomes in antiviral therapy, with a relatively low overall incidence of drug resistance. However, it is essential to enhance surveillance to reduce the spread of drug-resistant strains in the future.

Objective : To investigate the effect of lentivirus mediated silencing of NIPBL gene on osteogenic differentiation of mouse bone marrow mesenchymal stem cells ( BMSCs) . Methods : The third generation C57 Mouse Bone marrow mesenchymal stem cells were divided into experimental group , negative control group and blank control group. The lentiviral vector was transfected into mouse bone marrow mesenchymal stem cells , the transfection results were observed by inverted fluorescence microscope , and the expression of NIPBL gene was detected by real- time PCR. The cells of each group were cultured by osteogenic induction. The alkaline phosphatase activity was 2 and RUNX⁃2. Results : The expression of NIPBL mRNA decreased in the experimental group (P < 0. 05) . The activity of alkaline phosphatase in experimental group was lower than that in negative control group and blank control group (P < 0. 05) . The gene transcription and protein expression levels of OCN, BMP⁃2 and Runx⁃2 in experimental group were lower than those in negative control group and blank control group ( P < 0. 05 ) . Alizarin red staining results showed that the negative control group and blank control group had more red calcium nodules than the experimental group. Conclusion Lentivirus mediated silencing of NIPBL gene reduces the proliferation of mouse bone marrow mesenchymal stem cells , inhibits the expression of osteogenic differentiation related genes , and reduces the osteogenic differentiation ability.

Objective : To investigate the activation of the mammalian target of rapamycin complex 1 (mTORC1) signaling pathway molecules during the process by which kaempferol (Kae) promotes osteogenic differentiation of mouse bone marrow mesenchymal cells (BMMCs) under cyclic and uniaxial tension. Methods : BMMCs isolated and cultured in vitro were subjected to uniaxial dynamic tension with a 10% shape variable. The appropriate concentration of Kae was selected by cytotoxicity testing. The endogenous mTOR signal was inhibited by pp242. Four hours after traction, alkaline phosphatase (ALP) and osteocalcin (OCN) were detected by chemical colorimetry and ELISA, and the relative concentration of intracellular calcium was detected by flow cytometry. Phosphorylation of mTOR, 4E/BP1, and ribosomal protein S6 kinases (S6K), which are the main molecules of the endogenous mTORC1 signaling pathway, and expression of osteogenic transcription factors (Runx2 and Osterix) were detected by western blotting (WB), and mRNA expression levels of the above factors were detected by qRT-PCR. Results : The cytotoxicity test showed that 10 μmol/L Kae had little inhibitory effect on cell proliferation but had the strongest osteogenic ability. Four hours after stretching, Kae effectively promoted the osteogenic differentiation of BMMCs. The expression of ALP was （153.04 ± 18.72） U/mg, the expression of OCN was （1.64 ± 0.25） U. The mRNA and protein levels of Runx2 and Osterix were upregulated, and the intracellular calcium content was decreased. The mRNA and protein phosphorylation of mTOR and S6K was upregulated, and the opposite effect was observed with 4E/BP1. After pp242 was added to inhibit mTOR signaling, mTOR and S6K mRNA and protein phosphorylation were downregulated, but 4E/BP1 mRNA and protein phosphorylation was upregulated. The osteogenic differentiation of BMMCs was also significantly inhibited, mRNA and protein expression of Runx2 and Osterix were significantly downregulated, ALP and OCN expression were downregulated, and intracellular calcium content was increased. Conclusion Kae promotes osteogenic differentiation of mouse BMMCs under uniaxial dynamic tension through the mTORC1 signaling pathway.

Objective:To investigate the effect of AngioJet thrombectomy on proximal iliofemoral vein thrombosis after trauma.Methods:The clinical data of patients with proximal iliofemoral vein thrombosis treated with AngioJet thrombectomy were retrospectively analyzed at Vascular Surgery Department of our hospital in 2019.Results:A total of 76 patients were enrolled. After PMT 20 patients had grade Ⅲ thrombolysis clearance(26.3%), 41 patients (53.9%) had grade Ⅱ clearance, and 15 cases (19.7%) had grade Ⅰ clearance. Thirty four cases (44.7%) of grade Ⅲ were cleared after catheter aspiration or catheter thrombolysis, 35 cases (46.1%) of grade Ⅱ, 7 cases (9.2%) of grade Ⅰ were cleared, thrombus clearance rate increased significantly ( P<0.05). Cerebral hemorrhage occurred in 1 case (1.3%), and thrombosis recurred in 2 cases (2.6%). The patency rate was 91.8% at 3 months follow-up. Conclusions:AngioJet thrombectomy is a safe and effective method for treating post-traumatic proximal iliac femoral vein thrombosis. Intraoperative application of urokinase thrombolysis, catheter aspiration and catheter-directed thrombolysis can increase thrombus clearance effect.

Objective: To investigate the expression of the mTORC1 signaling pathway during the osteogenic differentiation of mouse bone marrow mesenchymal cells (BMMSCs) under cyclic uniaxial tension and explore its possible role. Methods : The BMMSCs of mice were affected by uniaxial dynamic tensile force. Western blot was used to detect the expression changes of major molecules (mTOR, Raptor, S6K) in the endogenous mTORC1 signaling pathway at 0, 1, 2, 4, and 8 hours after stretching. Chemical colorimetry, ELISA and PCR were used to detect alkaline phosphatase (ALP), osteocalcin (OCN) and Runx2 mRNA, respectively. Then, inhibition, activation and control groups were established by administration of the drugs PP242, MHY1485 and PBS, respectively. Two hours after the stress, the expression of S6K was detected by western blot, and the expression of the osteogenic signal was continuously detected by the above methods. Results : Western blot analysis showed that the main molecules of the mTORC1 signaling pathway were all expressed within 8 hours after traction, and the highest expression was 2 hours after the stress. Compared with those in the control group, the ALP activity and OCN expression decreased and the Runx2 mRNA levels increased after the mTORC1 signal pathway was inhibited (P ＜ 0.001); ALP activity and OCN expression increased after the mTORC1 signal pathway was activated, while the Runx2 mRNA levels decreased (P ＜ 0.001). Conclusion The mTORC1 signaling pathway participates in the osteogenic differentiation of mouse BMMSCs under tension. The osteogenesis of BMMSCs under cyclic uniaxial tension would be enhanced if the mTORC1 signaling pathway was activated.

Objective: To study the clinical features of dengue cases infected with hepatotropic virus and Mycobacterium tuberculosis in Xishuangbanna, and to provide evidences to set up effective treatment programs for the dengue patients infected with the other diseases for hospitals. Methods: The clinical characteristics of dengue cases infected hepatotropic virus and Mycobacterium tuberculosis were analyzed retrospectively on their symptoms and biochemical parameters from the People′s Hospital and the Infectious Disease Hospital of Xishuangbanna Prefecture in 2013 and 2015. Results: The clinical characteristics of dengue cases infected with hepatotropic virus were typical, and inclued low incidence of urinary abnormalities, coagulation disorders and high-lactate dehydrogenase. Dengue cases infected with Mycobacterium tuberculosis had high incidence of shock, high-hematocrit, renal function and coagulation abnormalities, which suggested a trend of more serious illness than other groups obviously. Conclusions The rate of severe disease was higher in dengue cases infected with Mycobacterium tuberculosis than those infected with hepatotropic virus, which suggests that the dengue cases infected with Mycobacterium tuberculosis should be treated timely to reduce the severity of the diseases in the hospital.

Objective : To explore the promoting effect of periostin on rapid distraction osteogenesis of the rabbit mandible and provide experimental evidence for the clinical use of periostin to promote osteogenesis. Methods: Twenty-four New Zealand male white rabbits underwent distraction osteogenesis, and after 3 days of retention, they were rapidly stretched at a stretch rate of 2 mm/d (total 5 d). The animals were randomly divided into group A and group B (12 per group). On the last day of the stretch, 0.5 mL of normal saline containing 40 μg of recombinant periostin was given to group B or an equal volume of normal saline was added to the control group (group A) for 8 days. At 4 weeks and 8 weeks post-stretch, 8 animals were randomly selected from each group to undergo a CT scan under general anesthesia. The bone mineral density and bone mineral content were detected by dual energy X-ray absorptiometry. Eight weeks post-stretch, all of the experimental animals were sacrificed. Six animals were randomly selected from each group for micro-CT and a histological examination, and the remaining animals were subjected to biomechanical tests. Results : CT images showed that the new bone formation in the distraction space of group B was significantly better than that of group A at 4 and 8 weeks post-stretch. At 4 weeks and 8 weeks post-stretch, the bone mineral density in group B was (0.157 ± 0.016) g/cm 2 and (0.234 ± 0.023) g/cm 2, respectively, and the bone mineral content was (0.096 ± 0.010) g and (0.204 ± 0.017) g, respectively. The above four means were significantly higher in group B than in group A (P < 0.001). The micro-CT images and data suggest that the stretch gap microstructure of group B has more mature features. Histological experiments showed that the trabecular bone of group B was thick and mature, with few chondrocytes. The biomechanical test results showed that the biomechanical strength of the distraction gap in group B was (228.47 ± 39.98) N, which was 1.24 times that of group A (P = 0.045). Conclusion Interstitial use of periosteal protein in the distraction space of the mandible in rabbits can promote local new bone formation and mineralization.

Objective To explore the expressions of growth factor receptor binding protein 2 -associated bin-ding protein -1 (Gab -1 )and glioma -associated oncogene homologue -1 (Gli -1 )in pediatric medulloblastoma,and to analyze their correlation between clinical and pathological characteristics and prognosis in pediatric medulloblastoma. Methods Elivision immunohistochemistry was used to detect the expressions of Gab -1 and Gli -1 protein in tissue microarray of 40 paraffin embedded pediatric medulloblastoma specimens.Chi -square test or Fisher exact test was used to analyze the correlation between Gab -1 and Gli -1 protein expressions with gender,age,tumor location and pathological subtypes.Follow -up data were handled by using Kaplan -Meier survival analysis and Cox regression anal-ysis.Results Positive expression ratios of Gab -1 and Gli -1 protein in 40 pediatric medulloblastoma were 35.0%and 55.0%,respectively.The positive expression rate of Gab -1 in medulloblastoma tissues had no statistical signifi-cance between different genders[male:30.4%(7 /23 cases)vs.female:41 .2%(7 /17 cases)],age[<3 years old:40.0%(6 /15 cases)vs.≥3 years old:32.0%(8 /25 cases)],tumor location[cerebellum:25.0%(5 /20 cases)vs. the fourth ventricle:45.0%(9 /20 cases)]and pathological subtype[classical type:40.7%(11 /27 cases)vs.desmo-plastic /nodular type:50.0%(5 /10 cases)vs.anaplastic /large cell type:66.7%(2 /3 cases)](χ2 =0.496,0.264, 1 .758,3.289,all P >0.05).There were statistical differences of positive expression rate of Gli -1 protein in different age groups[<3 years old:80.0%(12 /15 cases)vs.≥3 years old 40.0%(10 /25 cases)],different pathological sub-types[classical type:40.7%(11 /27 cases)vs.desmoplastic /nodular type:90.0%(9 /10 cases)vs.anaplastic /large cell type:66.7%(2 /3 cases)](χ2 =6.061 ,7.333,all P <0.05 ).There was no statistical difference in positive expression rate of Gli -1 protein between different gender[male:60.9%(14 /23 cases)vs.female:47.1 %(8 /17 cases)]and different tumor location [cerebellum:55.0% (11 /20 cases)vs.the fourth ventricle:55.0% (11 /20 cases)](χ2 =0.753,0.000,all P >0.05).Kaplan -Meier survival analysis showed that the age,the expressions of Gab -1 and Gli -1 protein were correlated with prognosis of pediatric medulloblastoma(all P <0.05).Cox regression indicated that the age,pathological subtypes and the expression of Gli -1 protein were independent prognostic indicators in pediatric medulloblastoma(all P <0.05).Conclusion Expression of Gab -1 and Gli -1 protein is significantly correlated with the prognosis of medulloblastoma,and the positive expression is a marker of unfavorable prognosis.

OBJECTIVETo understand the molecular characteristics of a dengue virus outbreak in China-Myanmar border region, Yunnan province, 2015 and provide etiological evidence for the disease control and prevention.

METHODSSemi-nested RTPCR was conducted to detect the capsid premembrane (CprM) gene of RNA of dengue virus by using dengue virus NS1 positive serum samples collected in Mengdin township, Gengma county, Yunnan province in July, 2015. Some positive samples were then detected by using PCR with specific primers to amplify the full E gene. The positive PCR products were directly sequenced. Then sequences generated in this study were BLAST in NCBI website and aligned in Megalign in DNAstar program. Multiple sequence alignments were carried out by using Mega 5.05 software based on the sequences generated in this study and sequences downloaded from GenBank, including the representative strains from different countries and regions. Phylogenetic trees were constructed by using Neighbor-Joining tree methods with Mega 5.05 software.

RESULTSTwenty one of 25 local cases and 10 of 14 imported cases from Myanmar were positive for DENV-1. Eight serum samples were negative for dengue virus. A total of 13 strains with E gene (1485 bp), including 8 local strains and 5 imported strains, were sequenced, which shared 100% nucleotide sequence identities. Twelve strains with CprM gene (406 bp) from 9 local cases and 3 imported cases shared 100% nucleotide sequence identities. Phylogenetic analyses based on E gene showed that the new 13 strains clustered in genotype I of dengue virus and formed a distinct lineage.

CONCLUSIONSThis outbreak was caused by genotype I of DENV-1, which had the closest phylogenetic relationships with dengue virus from neighboring Burma area. Comprehensive measures of prevention and control of dengue fever should be strengthened to prevent the spread of dengue virus.

Capsid Proteins ; China ; epidemiology ; DNA Primers ; Databases, Nucleic Acid ; Dengue ; epidemiology ; virology ; Dengue Virus ; genetics ; Disease Outbreaks ; Genotype ; Humans ; Myanmar ; epidemiology ; Phylogeny ; Polymerase Chain Reaction ; Sequence Alignment ; Software