Objective　To conduct an in-depth study of the molecular biological characteristics and evolutionary trends of H9N2 avian influenza virus (AIV) in live poultry markets in Yunnan Province in 2023, and to provide scientific evidence for the development of control strategies for H9N2 avian influenza in the region. Methods　Environmental samples were collected from live poultry markets in Yunnan Province in 2023 for H9N2 subtype nucleic acid detection. Positive samples were subjected to virus isolation using chicken embryos, and the genome of the 8 isolated strains was amplified, sequenced, and analyzed for genetic characteristics. Results　The eight avian influenza virus (AIV) isolates had the hemagglutinin (HA) cleavage site sequence PSRSSRGLF, which is a non-continuous basic amino acid sequence, consistent with the genetic characteristics of typical low-pathogenicity avian influenza viruses. Mutations Q234L and H191N were observed in the left arm of the HA protein, which enhanced the affinity for α-2,6 sialic acid receptors, suggesting that these viruses may have the potential to infect humans. The neuraminidase (NA) protein exhibited a deletion of three amino acids (TEI) at positions 62–64 in the stalk region, displaying characteristics of high pathogenicity at the molecular level. The increase or absence in potential glycosylation sites were observed in both HA and NA genes. The non-structural protein 1 (NS1) showed no D92E mutation, and had a C-terminal truncation of 13 amino acids, indicating that this virus is of low pathogenicity and poses a lower risk of human transmission. Mutations T37A, R95K, S224N, and K242N in the M1 protein of some isolates increased the risk of infection, while one isolate carried the V27A or S31N mutation in the M2 protein, conferring resistance to M2 ion channel inhibitors. Mutations M317I and S678N were identified in the PB1 protein, which may enhance pathogenicity in mice and increase the potential for mammalian infection. The PB2 protein carried the I292V mutation, which exhibited a stronger infectivity to mammals. Phylogenetic analysis revealed that the HA, NA, and PB2 gene segments belonged to the Y280 lineage, NP and PB1 gene segments were classified under the F98 lineage, the M gene segment of the NH013 isolate belonged to the F98 lineage, while M genes, as well as the NS and PA genes of other isolates belonged to the G1 lineage. Conclusions　These eight AIV isolates exhibited characteristics of low pathogenicity, but simultaneously carry the potential risk of infecting humans. Despite the HA cleavage site and NS1 protein mutations indicating low pathogenicity, the Q234L and H191N mutations in the HA protein enhanced its affinity for human receptors, suggesting the potential for human infection. The TEI deletion in the NA protein, mutations in the M1 protein, and resistance mutations in the M2 protein further increase the risk of human infection. Mutations in the PB1 and PB2 proteins increase the potential for these eight AIV strains to infect humans or mammals.

Objective To conduct a unique and pioneering molecular epidemiological investigation of a case of Eurasian avian-like H1N1 swine influenza identified in Yunnan Province in 2022(the fourth such case in the province)and to understand its genetic characteristics so as to reveal its potential impact on human health.Methods Real-time fluorescent quantitative PCR detection technology was used for the nucleic acid testing of the case's pharyngeal swab samples,close contacts,and environmental samples from the living area.Positive samples were subjected to virus isolation using MDCK cells.Cell cultures were authenticated using erythrocyte agglutination assay with guinea pig blood and real-time fluorescence quantitative RT-PCR.Whole genome sequencing was performed using the Illumina MiseqNext-generation sequencing platform,and a phylogenetic tree was constructed using MEGA 7.0 software to analyze the genetic molecular characteristics.Results The first G5 genotype Eurasian avian-like H1N1 swine influenza virus in Yunnan Province was successfully isolated,and the whole genome sequence of the virus was obtained.This virus possessed the molecular characteristics associated with increased adaptability,virulence,or transmissibility in mammals and had a nucleotide consistency of 99.2％～99.7％with a porcine strain isolated in Jiangsu province.These findings underscored the potential threat this virus poses to human health.Conclusion The study underscores the importance of further monitoring swine influenza in preventing new influenza virus subtypes that can infect humans.

Diabetes mellitus(DM)is a disease syndrome characterized by chronic hyperglycaemia.A long-term high-glucose environment leads to reactive oxygen species(ROS)production and nuclear DNA damage.Human umbilical cord mesenchymal stem cell(HUcMSC)infusion induces significant antidiabetic effects in type 2 diabetes mellitus(T2DM)rats.Insulin-like growth factor 1(IGF1)receptor(IGF1R)is important in promoting glucose metabolism in diabetes;however,the mechanism by which HUcMSC can treat diabetes through IGF1R and DNA damage repair remains unclear.In this study,a DM rat model was induced with high-fat diet feeding and streptozotocin(STZ)administration and rats were infused four times with HUcMSC.Blood glucose,interleukin-6(IL-6),IL-10,glomerular basement membrane,and renal function were examined.Proteins that interacted with IGF1R were determined through coimmunoprecipitation assays.The expression of IGF1R,phosphorylated checkpoint kinase 2(p-CHK2),and phosphorylated protein 53(p-p53)was examined using immunohistochemistry(IHC)and western blot analysis.Enzyme-linked immunosorbent assay(ELISA)was used to determine the serum levels of 8-hydroxydeoxyguanosine(8-OHdG).Flow cytometry experiments were used to detect the surface markers of HUcMSC.The identification of the morphology and phenotype of HUcMSC was performed by way of oil red"O"staining and Alizarin red staining.DM rats exhibited abnormal blood glucose and IL-6/10 levels and renal function changes in the glomerular basement membrane,increased the expression of IGF1 and IGF1R.IGF1R interacted with CHK2,and the expression of p-CHK2 was significantly decreased in IGF1R-knockdown cells.When cisplatin was used to induce DNA damage,the expression of p-CHK2 was higher than that in the IGF1R-knockdown group without cisplatin treatment.HUcMSC infusion ameliorated abnormalities and preserved kidney structure and function in DM rats.The expression of IGF1,IGF1R,p-CHK2,and p-p53,and the level of 8-OHdG in the DM group increased significantly compared with those in the control group,and decreased after HUcMSC treatment.Our results suggested that IGF1R could interact with CHK2 and mediate DNA damage.HUcMSC infusion protected against kidney injury in DM rats.The underlying mechanisms may include HUcMSC-mediated enhancement of diabetes treatment via the IGF1R-CHK2-p53 signalling pathway.

Objective:To study the correlation between myopia growth and physical development of primary school student in Shihezi area of Xinjiang Uyghur Autonomous Region by measuring the ocular biometry and physical parameters of the eye, and to provide an objective basis for the development of myopia prevention and control.Methods:A cross-sectional study design was used. The Grade 1-6 students from 33 schools in Shihezi area were randomly selected in October 2023. The children were classified according to age into the low age group (6-9 years) and the high age group (10-13 years).The children′s height, weight, axial length (AL), maximum keratometry (steepest keratometry, K1), and minimum keratometry (flattest keratometry, K1) were measured. The body mass index (BMI), keratometric astigmatism (ΔK), mean keratometry (K), corneal radius (CR) and AL/CR were calculated. Correlation analyses were performed between ocular biometry and physical parameters. LASSO regression analysis was used to screen the best variables, and linear regression models were constructed after adjusting for confounding factors of age and sex.Results:A total of 3 986 children aged 6-13 years were included in this study, including 2 094 boys (52.53%) and 1 892 girls (47.47%); 2 162 in the lower age group and 1 824 in the upper age group. Girls had higher K, ΔK and AL/CR values than boys (43.38 D vs 42.7 D, t=-7.84, P<0.01; 1.1 D vs 1.01 D, t=-2.14, P=0.03; 2.97 vs 2.96, t=3.32, P=0.02). Boys had higher AL, height, weight and BMI than girls (23.47 mm vs 23.02 mm, t=13.77, P<0.01; 1.34 m vs 1.33 m, t=3.02, P=0.03; 32 kg vs 29.5 kg, t=7.82, P<0.01; 17.4 vs 16.52, t=10.36, P<0.01). LASSO regression was used to screen the 3 physical parameter variables (height, weight, and BMI) to determine that height was the best physical parameter variable for AL/CR. After adjusting for confounders, it was shown that in the low age group (6-9 years), there was a significant positive correlation between children′s height and AL/CR ( b=0.174, 95% CI: 0.106 to 0.241, P<0.001), whereas in the high age group (10-13 years), there was no correlation between children′s height and AL/CR ( b=0.086, 95% CI:-0.038 to 0.21, P=0.174). Conclusion:There was a positive correlation between height and myopia growth in the lower age group of children in Shihezi area, Xinjiang Uyghur Autonomous Region; there was no correlation between height and myopia growth in the higher age group.

Objective:To study the correlation between myopia growth and physical development of primary school student in Shihezi area of Xinjiang Uyghur Autonomous Region by measuring the ocular biometry and physical parameters of the eye, and to provide an objective basis for the development of myopia prevention and control.Methods:A cross-sectional study design was used. The Grade 1-6 students from 33 schools in Shihezi area were randomly selected in October 2023. The children were classified according to age into the low age group (6-9 years) and the high age group (10-13 years).The children′s height, weight, axial length (AL), maximum keratometry (steepest keratometry, K1), and minimum keratometry (flattest keratometry, K1) were measured. The body mass index (BMI), keratometric astigmatism (ΔK), mean keratometry (K), corneal radius (CR) and AL/CR were calculated. Correlation analyses were performed between ocular biometry and physical parameters. LASSO regression analysis was used to screen the best variables, and linear regression models were constructed after adjusting for confounding factors of age and sex.Results:A total of 3 986 children aged 6-13 years were included in this study, including 2 094 boys (52.53%) and 1 892 girls (47.47%); 2 162 in the lower age group and 1 824 in the upper age group. Girls had higher K, ΔK and AL/CR values than boys (43.38 D vs 42.7 D, t=-7.84, P<0.01; 1.1 D vs 1.01 D, t=-2.14, P=0.03; 2.97 vs 2.96, t=3.32, P=0.02). Boys had higher AL, height, weight and BMI than girls (23.47 mm vs 23.02 mm, t=13.77, P<0.01; 1.34 m vs 1.33 m, t=3.02, P=0.03; 32 kg vs 29.5 kg, t=7.82, P<0.01; 17.4 vs 16.52, t=10.36, P<0.01). LASSO regression was used to screen the 3 physical parameter variables (height, weight, and BMI) to determine that height was the best physical parameter variable for AL/CR. After adjusting for confounders, it was shown that in the low age group (6-9 years), there was a significant positive correlation between children′s height and AL/CR ( b=0.174, 95% CI: 0.106 to 0.241, P<0.001), whereas in the high age group (10-13 years), there was no correlation between children′s height and AL/CR ( b=0.086, 95% CI:-0.038 to 0.21, P=0.174). Conclusion:There was a positive correlation between height and myopia growth in the lower age group of children in Shihezi area, Xinjiang Uyghur Autonomous Region; there was no correlation between height and myopia growth in the higher age group.

Abstract: Objective To investigate the molecular characteristics of the H9N2 avian influenza virus (AIV) causing human infection in Yunnan Province in 2019, and to provide the scientific basis for the prevention and control of avian influenza in Yunnan Province. Methods Influenza virus typing was performed by real-time RT-PCR in two influenza-like illness samples, and the Illumina Miseq high-throughput sequencer was used to determine the viral genome sequence. HA and NA gene sequence alignment and phylogenetic tree construction were performed using Mega7.0 software. Results Real-time RT-PCR results showed that two influenza-like illness samples were positive for H9N2 subtype. The full length of HA and NA were obtained by genomic sequencing. Sequence system evolution analysis showed that the HA and NA of the two AIVs in Yunnan Province were in the same evolutionary clade as A/Chicken/Zhejiang/HJ/2007 and belonged to the G57 type. The HA nucleotide and amino acid homology of the two AIVs were 93.92% and 95.00%, respectively, and the NA nucleotide and amino acid homology was 93.31% and 82.03%, respectively. The nucleotide (amino acid) homology of HA was 92.29%-96.94% (93.77%-98.43%) and 92.84%-94.92% (94.18%-96.23%), respectively, and NA nucleotide homology (amino acid) were 91.81%-97.60% (77.82%-94.83%), 94.38%-97.22% (85.47%-94.55%), respectively, compared with that of human infected H9N2 epidemic strains obtained in China from 2015 to 2020. Both AIVs HA protein cleavage site sequences were PSRSSR↓GLF, which was in line with the characteristics of low pathogenic influenza. The analysis of HA protein receptor binding site showed that amino acids at positions 109, 161, 163, 191, 202, 203 and 234 were consistent with the reference strains, while amino acids at position 198 were mutated to T. N166D and 168N mutations were also found in HA protein, and both AIVs had 7 potential glycosylation sites. Analysis of the erythrocyte binding site of NA gene found that there were amino acid mutations at positions 369, 402, 403, and 432, and amino acid deletion at positions 63-65 was found in the NA genes. There were 4 and 5 potential glycosylation sites in the two AIVs, respectively, and no drug resistance site mutations were found. Conclusions　The receptor binding sites, erythrocyte binding sites and glycosylation sites of HA and NA genes of H9N2 AIV in Yunnan Province have different degrees of variation, and monitoring and prevention and control should be strengthened.

Objective:To understand the characteristics of the local Coronavirus Disease 2019 (COVID-19) epidemic in the border areas of Yunnan province, and to analyze and trace the genome variation of the severe acute respiratory syndrome coronavirus 2 (SARS-COV-2).Methods:The basic information and samples of the first cases of 12 local epidemics in the border areas of Yunnan province from mid-February to April 2022 were collected. The virus genomes, genotypes and mutations were determined by the whole genome sequencing technology, the Pangolin COVID-19 Lineage Assigner and MEGA software. The sources of virus infection were estimated based on the contents of epidemiological investigations.Results:The first cases from 12 local epidemics in the border areas of Yunnan province had diverse occupations, and none of them had ever left the local area. The virus genomes of 12 first cases were Omicron (BA.2) variants and every two of them further belonged to the evolutionary clades BA.2.10 and BA.2.3.2, respectively. The genome-wide nucleotide-level mutation analysis showed that there were 1 or more specific nucleotide mutation sites in 10 first cases of the local epidemics. Combined with the epidemiological investigations, it was suggested that there might be different sources of virus infection in all 12 local epidemics. Further analysis of amino acid mutations revealed that in addition to the representative mutation sites of evolutionary clades, there were also specific amino acid mutation sites (such as S: E1188 D) in the genome sequences between the cases, which also deserved attention. The result of phylogenetic tree analysis were consistent with Pangolin Assigner and mutation analysis.Conclusions:The 12 local epidemics in Yunnan may have different sources of virus infection, which indicates that the COVID-19 epidemic in the border areas of Yunnan province is extremely complicated. Strengthening the frequency of nucleic acid screening and the scope of key populations and monitoring the variation of SARS-CoV-2 are of great significance for the precise prevention and control of the epidemic.

Objective:To investigate the clinical efficacy and safety of domestic bortezomib in the treatment of patients with multiple myeloma (MM).Methods:The data of 60 MM patients treated with domestic bortezomib as the basic chemotherapy regimen in 5 medical centers of Qilu Hospital of Shandong University, Heze Municipal Hospital, Weifang People's Hospital, Binzhou Medical University Hospital, Zibo Central Hospital in Shandong Province from January 2018 to June 2019 were retrospectively analyzed, 52 of which were newly treated patients and 8 were relapsed and refractory patients. The patients received at least 2 courses of combined chemotherapy based on domestic bortezomib, and the efficacy was assessed and evaluated every 2 courses.Results:Follow-up until June 30, 2019 showed that some patients were unable to return to the hospital for regular treatment. All patients completed at least 2 courses of treatment, with an overall effective rate (ORR) of 76.7% (46/60); 42 patients completed 4 courses of treatment, with an ORR of 78.6% (33/42); 30 patients completed 6 courses of treatment, with an ORR of 86.7% (26/30); there was no significant difference in ORR of 2, 4 and 6 courses ( P > 0.05). The complete remission+very good partial remission rates of 2, 4 and 6 courses were 16.7% (10/60), 47.6% (20/42) and 66.7% (20/30), respectively, and the difference was statistically significant ( P < 0.01). During the treatment, the adverse events mainly included infection, peripheral neuropathy, herpes, digestive tract symptoms, hematologic toxicities and so on, which were light and moderate mostly, and most of them can be reversed. The total incidence of adverse events in patients who completed 2, 4 and 6 courses of treatment were 91.7% (55/60), 66.7% (28/42) and 36.7% (11/30), respectively. Conclusions:The domestic bortezomib-based chemotherapy regimens have good efficacy in the treatment of MM. The incidence of adverse events is similar to that of the original drug, and patients can tolerate the adverse events.

Objective:To investigate the impact of recombinant human epidermal growth factor (rh-EGF) on the epithelial recovery and the tear film stability after trans-epithelial corneal collagen crosslinking in patients with progressive keratoconus.Methods:A randomized controlled clinical trail was designed.Consecutive 66 patients (37 males and 29 females) with an average age of (21.27±3.80) years old diagnosed with primary progressive keratoconus and hospitalized in Xiamen Eye Center Affiliated to Xiamen University from October, 2016 to January, 2017 were enrolled and treated with unilateral enhanced transepithelial corneal crosslinking surgery by iontophoresis, and the patients were randomly divided into control group and experimental group according to random number table method, with 33 patients 33 eyes in each group.The eyes in the control group were treated with carboxy-methylcellulose sodium lubricant eye drops and the eyes in the experimental group were treated with rh-EGF eye drops.The ocular surface disease index (OSDI) questionare, slit lamp examination, Schirmer Ⅰ test, corneal fluorescein sodium staining scoring, non-contact tonometry, uncorrected visual acuity, best corrected visual acuity, bulbar conjunctival congestion scoring, lacrimal sevretion test, non-invasive break-up time of tear film (NIBUT), as well as tear meniscus height analysis were performed before surgery, and on day 1, day 3, day 5, day 7, day 14 and day 28 after surgery.This study followed the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Xiamen Eye Center Affiliated to Xiamen University (No.2016-ME-003).Results:On day 7 after surgery, the OSDI values were increased in both groups than the preoperative value, while the value in the experimental group was significantly lower than that in the control group ( P<0.05). There were statistically significant differences in the overall corneal epithelial staining score values between the two groups at different time points ( Fgroup=16.701, P<0.01; Ftime=454.418, P<0.01). The corneal epithelial staining score in the experimental group on day 3 and day 5 after surgery were significantly lower than those in the control group (1.79±0.65 vs. 2.70±0.68; 0.91±0.46 vs. 1.55±0.51) (both at P<0.01). The conjunctival congestion score in the experimental group was significantly lower than that of the control group on day 3 and day 5 after surgery (both at P<0.05). There were statistically significant differences in the overall NIBUT values between the two groups at different time points ( Fgroup=13.084, P<0.01; Ftime=34.383, P<0.01). The NIBUT values were significantly decreased rapidly on day 7, day 14 and day 28 after surgery in both groups (all at P<0.01), but gradually recovered.The NIBUT of the experimental group on day 7 and day 14 after surgery were significantly higher than those of the control group ([8.18±2.26]seconds vs. [5.93±2.33]seconds; [9.49±1.95]seconds vs. [7.52±2.27]seconds) (both at P<0.01). No statistical differences were found in the tonometry value, visual acuity, value of Schirmer I test as well as tear meniscus height at any time point before or after surgery between the two groups (all at P>0.05). Conclusions:Recombinant human epidermal growth factor has positive effects in the patients received enhanced transepithelial corneal crosslinking surgery, presenting with promotion of epithelial healing, relief of post-operative discomfort, and the recovery of tear film stability.

Objective:To understand the epidemiological characteristics of an outbreak of Chikungunya fever (CHIK) in Ruili city at Sino-Myanmar border area in Yunnan province, China, 2019.Methods:The data of CHIK cases in Ruili city were collected. Serum samples were collected from acute phase of the patients for the detection of the viral nucleic acid with real-time reverse transcription PCR assay.Results:A total of 121 CHIK cases (laboratory-confirmed cases) were reported in Ruili city in 2019, in which 98 were indigenous cases (80.99%) including 2 pregnant women and 3 neonatal cases (twin boys and 1 girl) were observed with vertical mother-to-child transmission), and 23 were imported cases (19.01%) from Myanmar. CHIK mainly occurred in Urban area (Mengmao town) and port area (Jiegao development zone) of Ruili city. The epidemic of CHIK lasted from September to November. The epidemic peak was in October (74.38%). The majority of patients were aged from 20—59 years (77.69%), in addition to 3 cases of mother-to-child transmission, the youngest was 3 months and the oldest 72 year-old. The male to female ratio was 1∶1.28 (53∶68). Occupational distribution of CHIK cases was mainly for the commercial service personnel (33.06%), home workers and unemployed persons (21.49%) and retiree (11.57%).Conclusions:During the outbreak of CHIK in Ruili in 2019, both indigenous and imported cases were detected. The imported CHIK cases from Myanmar were the main sources for local transmission of CHIK in Ruili city. This is the first time that a local epidemic of CHIK has occurred in Yunnan Province. It is necessary to strengthen control of the cross-border spread of CHIK in this area.