Objective To investigate the effects of exposure to various concentrations of cigar smoke on gut microbiota in mice.Methods A total of 40 C57BL/6 mice were randomly divided into the control group,the low-dose cigar exposure group,the medium-dose group and the high-dose group,with 10 mice in each group.After 4 weeks of feeding,fecal samples were collected for gene sequencing of 16S ribosomal RNA and analysis of differences in gut microbiota.Results Compared to the control group,gut microbiota richness was signifi-cantly reduced in the cigar-exposed groups(P<0.05).Compared with thecontrol group,the Shannon index of mice in the high-dose group was significantly increased(P<0.05).In multi-group comparisons,ten bacterial genera with high abundance-such as Akkermansia,Allobaculum,and Alloprevotella-were identified.Pairwise comparison results indicated that compared to the control group,abundances of Akkermansia,Candidatus_Sac-charimonas,and Lactobacillus decreased while those of Allobaculum,Alloprevotella,Muribaculaceae,and Pre-votellaceae_UCG001 increased(P<0.05).Alistipes and Faecalibaculum showed significant increases in low-dose and medium-dose groups respectively,Blautia and Lachnospiraceae_NK4A136 group exhibited notable in-creases in the high-dose group(P<0.05).Linear discriminant analysis effect size revealed that six phyla and forty-four species displayed significant differences across all groups at both phylum and species levels,distinct dose-specific were observed among different cigar exposure groups.Conclusion Cigar smoke exposure and different exposure concentrations can both cause changes in the gut microbiota.The effects of different con-centrations of cigars on the gut microbiota of mice are specific.

Objective:To analyze the genetic and molecular characteristics of H9N2 subtype avian influenza viruses from external environment and humans in Anhui Province from 2013 to 2022.Methods:Environmental samples and human samples were collected from Anhui influenza surveillance network laboratory. Sixty-three strains of virus were isolated in chicken embryos. RT-PCR was used to amplify the virus and whole genome sequencing was performed. To construct gene evolutionary tree and analyze its genetic characteristics and potential glycosylation sites.Results:The hemagglutinin (HA) gene belongs to the 9.2.4.5 clade, and the protein cleavage sites are mostly " PSRSSR\GL". The neuraminidase (NA) gene, basic protein-1(PB1) gene, acidic protein (PA) gene, non-structural protein (NS) gene and nucleoprotein (NP) gene belong to the F/98 clade, the matrix protein (MP) gene belongs to the G1/97 clade, and the basic protein-2 (PB2) gene belongs to the ST/7488 clade. Mutations of T155N, R164Q, H183N, T189D/V, A190V/T and Q226L occurred in HA protein, deletion of NA protein occurred at 62-64 sites, and mutations of T271A, I292V/M and E627V/L occurred in PB2 protein. At the same time, mutations of K356R and S409N occurred in the PA protein.Conclusions:The H9N2 subtype avian influenza viruses collected from external environment and human sources in Anhui province from 2013 to 2022 belong to the same evolutionary branch, and amino acid site mutations suggest that the virus shows a tendency to gradually adapt to the mammalian host environment. Therefore, further studies on the adaptive evolution of the virus and related monitoring work are needed.

Objective:To explore the structural characteristics of intestinal microflora in children with non-alcoholic fatty liver disease（NAFLD）and the relationship between intestinal microflora and the occurrence as well as development of NAFLD in children.Methods:Fifteen children with NAFLD diagnosed at the First People's Hospital of Yunnan Province from January 2022 to December 2022 were selected as subjects，and 15 healthy children who received routine physical examinations at the outpatient clinic during the same period were randomly selected as healthy control group.The height，weight，waist circumference，blood pressure，blood biochemistry of all children were collected.At the same time，the fresh fecal samples of all children were collected，and the biological information of intestinal flora was analyzed by 16S rRNA sequencing.Results:In the NAFLD group，there were eight males and seven females，with an average age of（11.13±1.77）years.In healthy control group，there were seven males and eight females，with an average age of（9.73±2.25）years.There were no significant differences in sex，age，blood pressure between two groups.Compared with the healthy control group，the levels of body mass index，waist circumference，waist-to-height ratio，alanine aminotransferase，aspartate transaminase，gamma-glutamyl transpeptidase，total bilirubin，direct bilirubin，unconjugated bilirubin，low density lipoprotein cholesterol，uric acid and serum insulin significantly increased and high density lipoprotein cholesterol significantly decreased in NAFLD group（ P<0.05）.The results of species diversity analysis showed that chaol index and observed-species index in NAFLD group were significantly higher than those in healthy control group（ P<0.05）.Species diversity analysis showed that the species with increased abundance in NAFLD group included：Proteobacteria，Gammaproteobacteria，Enterobacteriaceae，Klebsiella，Escherichia-Shigella，Escherichia-Shigella-unclassified.Differential species with increased abundance in the healthy control group included：Bifidobacterium species，Bifidobacterium，Bifidobacteriaceae，Bifidobacteriales，Actinobacteria，Actinobacteriota，Bacteroidia，Bacteroidales，Streptococcus-thermophilus. Conclusion:There are metabolic abnormalities and obvious changes in the structure of intestinal flora in children with NAFLD.Exogenous supplementation of Bifidobacterium，Streptococcus thermophilus and Bacteroides may prevent the occurrence of NAFLD，delay the progression of disease and improve fat deposition in the liver.

Objective:To analyze the protective effect of weight-loss based exercises on the knee joints of patients with simple obesity.Methods:This study was a randomized controlled trial, 60 patients with simple obesity who met the inclusion and exclusion criteria and treated at the Affiliated Panyu Central Hospital of Guangzhou Medical University from April 2021 to April 2022 were selected as subjects. The patients were randomly divided into a power bicycle group (weight-loss, 30 cases) and a moving treadmill training group (non-weight-loss, 30 cases) using the random envelope method. High intensity interval exercise training was conducted in the hospital: slow warm-up for 1 minute, followed by rapid exercise for 3 minutes, and then rest for 1 minute. Repeated this cycle twice to form one set, and to perform 3 sets per session, 5 times per week, for a continuous training of 12 weeks. In addition, patients were not allowed to participate in other sports activities, and they should maintain their original dietary habits and patterns unchanged. Body mass index and body fat percentage were measured before and after treatment, and the cases with knee pain were recorded. The volume of knee cartilage was measured using magnetic resonance imaging. The local skin temperature of the knee was measured using an electronic thermometer before and after each training session. The t-tests, Pearson-tests and Mann-Whitney U tests were used to compare the differences in the above indicators between two groups before and after intervention to analyze the protective effect of weight-loss based exercise on the knee joints of patients with simple obesity. Results:There was no statistically significant difference in baseline data such as gender, age, height, and weight between the two groups before the intervention (all P>0.05). After the intervention, the body mass index, percentage of body fat, and knee cartilage volume of both groups were significantly lower than before the intervention, and the incidence of knee pain and local skin temperature of knee were significantly higher than before the intervention (all P<0.001). The incidence of the knee pain, reduction in knee cartilage volume, and elevation in local skin temperature in the power cycling group were all significantly lower than those in the moving treadmill training group [16.67% vs 100%, 0.12 (0.05, 0.27) vs 1.68 (1.15, 2.21) cm 3, (0.17±0.09) vs (0.64±0.15)℃, respectively] (all P<0.001). And there was no statistically significant difference in body mass index and body fat percentage between the two groups (both P>0.05). Conclusion:The weight-loss based exercise has a protective effect on the knee joint of patients with simple obesity, which can avoid a reduction in volume of knee cartilage and elevation in local skin temperature, and reduce the occurrence of pain.

Objective To develop an asymmetric porous guided bone regeneration(GBR)membrane made from antibacterial zinc oxide nanoparticle(nZnO)/poly(3-hydroxybutyrate-co-4-hydroxy butyrate)copolymer(P34HB),and detect its osteogenic performance.Methods Ultrasonic dispersion technique,solvent displacement method and freeze-drying technique were used to prepare P34HB composite GBR membrane loaded with nZnO at different doses(0％,0.5％,1.0％and 1.5％).Scanning electron microscopy(SEM)and energy spectrometry(EDS)were conducted to determine the load of nZnO and observe the structure of the prepared membrane.Then MC3T3-E1 cells were implanted in a 24-well culture plate with the prepared GBR membrane pre-laid in the well.Thus the experiment included blank control group(no membrane)and groups with membranes composing 0％,0.5％,1.0％and 1.5％nZnO,respectively.CCK-8 and cell scratch assays were used to detect cell proliferation capacity and migration ability.The protein adsorption capacity of GBR membrane was detected by protein adsorption experiment.The in vitro degradation of GBR membrane was tested;the activity of alkaline phosphatase(ALP)was detected with chemical test.And calcified nodules were observed with alizzarin red staining;the antibacterial ability of GBR membranes was tested with in vitro antibacterial experiments.Results Four groups of nZnO/P34HB composite GBR membranes with 0％,0.5％,1.0％and 1.5％nZnO were successfully prepared.SEM results showed that the composite membrane was porous on one side and dense on the other side.EDS indicated the incorporation of zinc elements in the composite membrane.The results of cell proliferation assay showed that 0.5％and 1.0％nZnO/P34HB group promoted the proliferation of MC3T3-E1 cells,when compared with the blank control group and 0％and 1.5％nZnO/P34HB groups(P＜0.05).The results of cell scratch assay showed that MC3T3-E1 cells in the 0.5％nZnO/P34HB group migrated significantly over the scratch edge,with the cell mobility higher than that of the other 3 membrane groups in 48 h after culture(P＜0.01).Protein adsorption experiment displayed there was no significant difference in optical density of the 4 group of membrane,indicating that the addition of nZnO had no effect on the protein adsorption capacity of the material itself.The results of enzymatic experiment showed that though the quality of the 4 membranes was continuously decreased with elapse of time,the degradation rate was still relatively lower after 28 d of treatment.ALP staining and alizarin red staining displayed that the 0.5％nZnO/P34HB group obtained the strongest ALP activity and the largest amount of calcified nodules when compared with the other 3 groups(P＜0.01).Higher mineralization was observed in the 0％and 0.5％nZnO/P 34 HB groups than the other 2 groups(P＜0.01).The results of in vitro antibacterial experiments showed that the number of colonies in the 0.5％nZnO/P 34 HB group was significantly less than that in the blank control group and the 0％nZnO/P34HB group(P＜0.01).Conclusion Our prepared P34HB composite GBR membrane loaded with nZnO is in asymmetric porous structure,and can promote cell proliferation,facilitate cell osteogenesis and have bacteriostatic ability.

Objective To investigate the clinical features of the spectrum of upper gastrointestinal diseases in children and to provide assistance in the prevention,diagnosis and treatment of upper gastrointestinal diseases in children.Methods Clinical data of 424 children aged 2 months to 14 years who were hospitalised for gastroscopy in the paediatrics department of the First People's Hospital of Yunnan Province from January 2018 to June 2023 were collected,and retrospective analyses were performed for the children's clinical presentation,gastroscopy results,and HP infections.Results The symptoms of vomiting and diarrhea in infants were significantly different from those in preschool and school age groups(P＜0.05).The positive rate of lesions under gastroscopy was 96.46％.The main diseases found were non-atrophic gastritis(222 cases),bile reflux gastritis(42 cases),non-atrophic gastritis with erosion(30 cases),duodenitis(25 cases),reflux esophagitis(15 cases)and others(75 cases).Helicobacter pylori(HP)was detected in 300 of the 424 cases,and 105 cases were HP positive,with a detection rate of 35.00％.Conclusion Abdominal pain was the main symptom of upper gastrointestinal diseases in school-age and pre-school children.Vomiting and diarrhea were the main symptoms of upper gastrointestinal diseases in infants.There was no gender difference in HP infection,but there was a difference in age.

【Objective】 To estimate the prevalence, associated factors and patterns of multimorbidity of non-communicable diseases among adults in Shaanxi Province so as to provide evidence for the prevention and control of non-communicable diseases. 【Methods】 We used the data of adults aged 18 years and older collected in the baseline survey of Shaanxi Project in the Regional Ethnic Cohort Study in Northwest China. Multinomial logistic regression was used to explore the associated factors for multimorbidity. Exploratory factor analysis was used to extract patterns of multimorbidity. 【Results】 The prevalence of multimorbidity was 10.7% among the 44 442 participants. Age increase, being males, urban residence, and being overweight or obesity were positively associated with multimorbidity. Compared with women, men had a higher risk of multimorbidity. The OR and 95% CI was 1.25 (1.12-1.39). The risk of multimorbidity increased with age among adults. Compared with participants aged 18.0-34.9 years, the ORs and 95% CIs of those aged 35.0-44.9, 45.0-54.9, 55.0-64.9, and ≥65.0 years were 4.73 (3.47-6.46), 15.61 (11.60-21.00), 41.39 (30.76-55.70) and 90.04 (66.58-121.77), respectively. The primary multimorbidity patterns among adults in Shaanxi were cardiovascular-metabolic multimorbidity (5.4%), viscero-articular multimorbidity (1.0%), and respiratory multimorbidity (0.3%). 【Conclusion】 More than one in ten adults in Shaanxi Province had multimorbidity, and the predominant pattern of multimorbidity was cardiovascular-metabolic multimorbidity. The prevention and control of non-communicable diseases should be reinforced in middle-aged and older people, males, people living in the urban, and overweight or obese people. More attention should be paid to the prevention and control of cardiovascular-metabolic diseases.

【Objective】 To evaluate the dietary quality with the dietary balance index (DBI_16) and the association between dietary quality and bone mass among middle-aged and elderly people in Gansu Province so as to provide evidence for improving dietary quality and bone health status of Gansu population. 【Methods】 Based on the information of the type and quantity of food intake and the bone mass of middle-aged and elderly people aged 35 years and above collected by the Gansu Project in the Regional Ethnic Cohort Study in Northwest China, DBI_16 was used to evaluate the intake level of cereals, vegetables, fruits, milk, beans, fish and shrimp, eggs and other foods, and the degree of inadequate, excessive and unbalanced dietary intake of the participants. Multiple linear regression was used to evaluate the associations of three component indexes of DBI_16, high bound score (DBI_HBS), low bound score (DBI_LBS), diet quality distance (DBI_DQD), and seven single indexes of DBI_16 with bone mass. 【Results】 Analyses of the dietary and bone mass data of 11,840 participants showed that 44.8% of participants consumed excessive amounts of cereals compared to the dietary recommendation. 96.3%, 90.6%, 90.1%, 71.9%, 95.1% and 60.3% of participants’ intake of vegetables, fruits, milk, soybeans, fish and shrimp, and eggs, respectively, were inadequate. 47.7% participants consumed less than 10 types of food. 2.3% participants’ DBI_LBS levels were appropriate. 54.7% participants’ DBI_HBS levels were appropriate. Only 1.2% participants’ DBI_DQD reached a balanced level. The bone mass level in the study population was (2.5±0.6) kg [(2.8±0.5) kg for men and (2.3±0.5) kg for women]. After adjusting for sociodemographic characteristics, lifestyle, total dietary energy intake and body mass index, DBI_LBS and DBI_DQD were negatively associated with bone mass [β and 95% CI was -0.002 01 (-0.003 62--0.000 40) and -0.001 76 (-0.003 09--0.000 43), respectively]. 【Conclusion】 Dietary intake imbalance is common among middle-aged and elderly people in Gansu Province, and the more severe the dietary intake imbalance, the lower the bone mass level.

Objective:To compare the differences between suspension and adherent cells of MDCK cell line in the isolation of influenza virus, and to explore the application prospects of MDCK cell suspension.Methods:Determination of viable cell density and cell specific growth rate were recorded by cell count. The WHO recommended vaccine strains were used for virus infection experiments. After five passages, hemagglutination titers were detected, while the sequencing analysis of their HA and NA genes revealed the mutation frequency.Results:The 24-hour and 48-hour viable cell density of the cell suspension was more stable than that of adherent cells. The cell suspension achieved an HA titer of 1∶256 or higher in the third generation, while adherent cells had no titer. In the fourth and fifth generations, one amino acid site mutation was found in the HA gene of H3N2 and BV subtypes of influenza virus cultured in the cell suspension, while no gene mutation was found in adherent cells in two passages. There were no mutations in the whole NA gene.Conclusions:Suspension of MDCK cells have more stable growth and higher efficiency in virus isolation than adherent cells, meanwhile there was a low rate of virus mutation during continuous passage. This study demonstrated the feasibility of this suspension of MDCK cells for influenza vaccine production based on cell culture technology.

Objective:Analysis of the characteristics of influenza epidemic in Anhui Province and quantification of the impact of different factors on influenza occurrence, providing scientific basis for better influenza prevention and control.Methods:Descriptive analysis and factor analysis were conducted on influenza-like illness (ILI) cases and RT-PCR results in Anhui Province from 2013 to 2021 using data from China's Influenza Monitoring Information System.Results:The percentage of influenza-like illness (ILI%) of sentinel hospitals in Anhui Province from April 1, 2013 to March 31, 2021 was 3.80% (1 209 142/31 779 987), showing an overall increasing trend, with a relatively high proportion in 2017-2018 at 4.30% (191 148/4 448 211). The proportion of ILI cases in infants and young children aged 0-4 years was a relatively high at 54.14% (654 676/1 209 142), and the highest ILI% was observed in Fuyang City, Anhui Province (6.25%, 236 863/3 788 863). Laboratory monitoring results showed that the positive rate of ILI cases in sentinel hospitals in 8 influenza monitoring years was 16.38% (34 868/212 912), showing an increasing trend year by year, with a relatively proportion in 2017-2018 at 26.19% (6 936/26 488). The detection rate of school-age children aged 5-14 years was a relativelyhigh at 28.81% (13 869/48 144), and the positive rate was a relatively high in Wuhu City among the 16 cities, reaching 22.01% (2 693/122 237). Influenza activity showed a single peak in winter-spring and alternating double peaks in winter-spring and summer, with different subtypes alternating, and A (H3N2) was the dominant subtype in summer. The results of a multiple logistic regression model showed that the positive rate was higher in 2017-2018, among children aged 5-14 years, in winter, and in southern Anhui.Conclusions:Influenza epidemic in Anhui Province has a clear seasonal pattern, and the ILI% and detection rate have shown an upward trend from 2013 to 2021. Therefore, it is suggested to ensure vaccine supply before the winter-spring influenza season arrives, and to strengthen vaccine uptake and health education to avoid the risk of infection during the peak period of influenza.