Objective: To investigate the trend in incidence of stroke in Yongkang City, Zhejiang Province from 2015 to 2022, so as to provide the evidence for improving stroke prevention and treatment strategies. Methods: The incidence of stroke in Yongkang City from 2015 to 2022 was collected through the Zhejiang Chronic Disease Monitoring Information System, and standardized by the data of the Chinese National Population Census in 2010. The incidence of stroke in gender-, age- and subtype-specific was analyzed, and the incidence trend of stroke was analyzed by average annual percentage change (AAPC). Results: A total of 22 408 stroke cases were reported in Yongkang City from 2015 to 2022, with crude incidence of 457.97/105 and standardized incidence of 379.55/105. The crude incidence of stroke appeared a tendency towards a rise (AAPC=6.447%, P<0.05). The crude incidence of stroke was higher among males than females (495.97/105 vs. 417.58/105, P<0.05), and the standardized incidence of stroke was higher among males than females (425.29/105 vs. 332.49/105, P<0.05). The average age of stroke cases was (70.74±12.64) years. The crude incidence of stroke appeared a tendency towards a rise with age (P<0.05), and which in residents aged 30 to 44 years showed an increasing trend (AAPC=6.142%,P<0.05). There were 18 040 cases of ischemic stroke (80.51%), 4 121 cases of hemorrhagic stroke (18.39%) and 247 cases of unclassified stroke (1.10%) reported from 2015 to 2022. The crude incidence of ischemic stroke was higher than which of hemorrhagic stroke (368.70/105 vs. 84.22/105, P<0.05). The crude incidence of ischemic stroke appeared a tendency towards a rise (AAPC=7.851%, P<0.05). Conclusions The crude incidence of stroke appeared a tendency towards a rise in Yongkang City from 2015 to 2022, with ischemic stroke as the predominant subtype. Male and the elderly should be given a high priority for stroke control.

This study was aimed to investigate the effect of Yi-Qi Wen-Yang (YQWY) and Huo-Xue Li-Shui (HXLS) decoction on changes of insulin resistance (IR) model and the myocardial tissues of rats with congestive heart failure (CHF) pathology. CHF rat models were established on Wistar male rats through injection of doxorubicin hydrochlo-ride into rat tail vein. Wistar male rats models were randomly divided into the model group, western medicine group, low dose decoction group, middle dose decoction group, and high dose decoction group. After 4-week gavage, 3 mL vein blood was taken from the angular vein sinus for the determination of blood glucose and serum insulin, and the calculation of IR. Finally, the rats were sacrificed. And then, the heart was removed to make HE slice and observe the pathological change of myocardium. The results showed that compared with the model group, YQWY and HXLS decoction can improve the IR level among CHF rats (P<0.05). Among them, the effects of the high dose and middle dose group were obvious. At the same time, this decoction can improve the myocardial cells in CHF rats in myocar-dial cells of the high dose group. And its morphology change was close to the digoxin group. It was concluded that YQWY and HXLS decoction can reverse IR and improve ventricular remodeling among CHF rats to a certain extent.

This study was aimed to observe the protective effect of astragalus on bones of vitamin D deficiency rat model. A total of 30 male SD rats were randomly divided into 3 groups, which were the control group, model group, and the astragalus group. The experimental period was 8 weeks. After the experiment, enzyme immunoassay was used for the detection of serum 25 hydroxy vitamin D3 [25 (OH)VD3], fibroblast growth factor-23 [FGF-23], Klotho, and HE staining of femur. The results showed that compared with the normal control group, the vitamin D deficiency rat model group had a decrease in both serum 25 (OH)VD3 and Klotho, and a increase in FGF-23, which meant the ex-istence of osteoporosis. Compared with the model group, the astragalus group had a decrease in both the serum 25 (OH)VD3 and FGF-23, and a increase in Klotho, which meant the osteoporosis of the astragalus group had been im-proved significantly. It was concluded that astragalus can regulate FGF-23 and Klotho in vitamin D deficiency rats in order to have a protective effect for bones.

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 evaluate the influence of preconditioning with and anti-myosinmonoclonal antibody (mAb2G4)-nuclear factor-kappa B decoy oligodeoxynucleotide (ODN)-lipofectamine (lip) on hypoxia-reoxygenation (H/R) injury in H9c2 cardiomyocytes.Methods H9c2 cardiomyocytes were seeded in 6-well plate at the density of 1×105/ml (2 ml/well),and were divided into 3 groups (n=9 each) using a random number table:control group (group C),H/R group and mAb2G4-ODN-lip group (group MOL).The cells underwent 2 h of hypoxia in an air-tight bag,followed by 1 h reoxygenation.In MOL group,the cells were treated with mAb2G4-ODN-lip (2 μg ODN) for 4 h and then cultured in the common culture medium for 8 h before hypoxia.At the end of reoxygenation,proliferation of cells was measured using MTT assay,and the cells and supernatant of the culture medium were collected to determine the activity of lactate dehydrogenas (LDH),content of malondialdehyde (MDA),concentrations of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) (by ELISA).The rate of proliferation inhibition was calculated.Results Compared with group C,the rate of proliferation inhibition,LDH activity,MDA content,and concentrations of TNF-α and IL-6 were significantly increased in the other two groups.Compared with group H/R,the rate of proliferation inhibition,LDH activity,MDA content,and concentrations of TNF-α and IL-6 were significantly decreased in MOL group.Conclusion mAb2G4-ODN-lip can mitigate H/R injury in H9c2 cardiomyocytes.

Th22,a new subset of helper T cells,which is char-acterized by the secretion of interleukin-22(IL-22),could infil-trate to the epidermis in individuals with inflammatory skin disor-ders.This article introduces the action of Th22 and IL-22 in in-flammatory skin diseases,including psoriasis,atopic dermatitis, systemic lupus erythematosus,systemic sclerosis,aiming at re-vealing the role of Th22 and IL-22 in these diseases,which would not only provide some novel targets of drugs for inflamma-tory skin diseases,but also promote the researches on the pre-vention and treatment of these diseases.

Objective To evaluate the effect of oxycodone preconditioning on liver injury induced by intestinal ischemia-reperfusion (I/R) in rats and the role of different opioid receptors.Methods Fiftyfour adult male Sprague-Dawley rats, weighing 200-300 g, were randomly divided into 9 groups (n =6 each) using a random number table: sham operation group (group S), group I/R, oxycodone preconditioning group (group OP) , μ receptor antagonist CTOP group (group CTOP) , δ receptor antagonist naltrindole group (group NTD), κ receptor antagonist nor-binaltorphimne group (group BNI), CTOP + oxycodo ne preconditioning group (group CTOP+OP) , naltrindole + oxycodone preconditioning group (group NTD+ OP) , and nor-binaltorphimne + oxycodone preconditioning group (BNI+OP).The model of intestinal I/R was established by occlusion of the superior mesenteric artery for 45 min followed by 2 h reperfusion in anesthetized rats.The superior mesenteric artery was only exposed, but not occluded in group S.In OP,COTP+OP, NTD+OP and BNI+OP groups, oxycodone 0.5 mg/kg was injected intravenously at 10 min prior to ischemia.COTP 1 mg/kg and naltrindole 5 mg/kg were injected intravenously at 20 min prior to ischemia in COTP+OP and NTD+OP groups, respectively.Nor-binaltorphimne 5 mg/kg was injected intravenously at 25 min prior to ischemia in group BNI+OP.In CTOP and NTD groups, the corresponding doses of CTOP and naltrindole were injected intravenously at 10 min prior to ischemia.In group BNI, the corresponding dose of nor-binaltorphimne was injected intravenously at 15 min prior to ischemia.The rats were sacrificed at 2 h of reperfusion, and left hepatic lobes were removed for microscopic examination and for detection of apoptosis in liver cells (using TUNEL).The apoptosis index (AI) was calculated.Results Compared with group S, the AI was significantly increased in the other groups (P＜0.05).Compared with group I/R, the AI was significantly decreased (P＜0.05) , and the pathological changes of livers were reduced in OP, COTP+OP, NTD+OP and BNI+OP groups, and no significant change was found in AI and pathological changes of livers in CTOP, NTD and BNI groups (P＞0.05).Compared with group OP, the AI was significantly increased (P＜0.05), and the pathological changes of livers were aggravated in COTP+ OP, NTD+OP and BNI+OP groups.There was no significant difference in AI and pathological changes of livers among groups COTP+OP, NTD+OP and BNI+OP (P＞0.05).Conclusion Oxycodone preconditioning can mitigate liver injury induced by intestinal I/R in rats, and μ, δ and κ receptors mediate the role with comparable effects.

Objective: To obtain monoclonal antibodies against programmed cell death 10(PDCD10) for further study of the structure and function of PDCD10 protein.Methods: Balb/c mice were immunized with recombinant PDCD10,hybridoma cell lines secreting monoclonal antibodies against PDCD10 were screened by regular cell fusion and subcloning approach.The specificities of these monoclonal antibodies were determined by ELISA,Western blotting and Immunofluorescecence assay.Results: Three hybridoma cell lines(5G1,4F7 and 3H5) stable in secreting specific monoclonal antibodies were successfully obtained.Subclass of IgG belonged to IgG1(4F7 and 5G1)and IgG2b(3H5),respectively.The ascite titers of these monoclonal antibodies reached 1∶10~7.They could specifically bind to recombinant PDCD10 and endogenous and overexpressed PDCD10 proteins proved by ELISA and Western blotting.They failed to react with E.coli lysates and glutathione S-transferase(GST).In addition,these three monoclonal antibodies could recognize different epitopes of PDCD10 proteins assessed by immune fluorescence competitive binding assay.Both endogenous and overexpressed PDCD10 protein mainly located in the nucleus.Conclusion: Monoclonal antibodies against PDCD10 with high titers and specificity have been successfully prepared,which has laid the foundation for further study of PDCD10 protein.

Long non-coding RNA (lncRNA) is a newly identified non-coding RNA subfamily with various regulatory functions.Recent evidence has shown the fundamental role of long noncoding RNAs in affecting the development of diseases at different levels,from gene modification,transcriptional regulation to protein transla tion.The study on lncRNA has made great progress in the studies of genomic imprinting,cancer diseases and neurodegenerative disorders,while the research relevant to autoimmune diseases has just staaed recently,However,many lncRNAs have been identified to involve in immune cells proliferation,differentiation and maturation,acting as the key regulators in immune homeostasis and autoimmune diseases.This review is focused on the advances of lncRNA in immune cells and autoimmune diseases.

Objective To observe the expression of discs large hom olog 4 (DLG4) protein in hippocam-pus, am ygdala and frontal cortex of rats and evaluate postsynaptic density in heroin dependence. Meth-ods The rat heroin dependent m odel was established by increasing intraperitoneal injection of heroin. DLG4 proteins in hippocam pus, am ygdala and frontal cortex of heroin dependent 9, 18, 36 days rats w ere detected with im munohistochem ical staining and com pared with that in the control group. Results DLG4 proteins in hippocam pus, am ygdala and frontal cortex w ere gradually reduced with extension of heroin dependent tim e. Conclusion Heroin dependence can affect postsynaptic density of hippocam pus, am ygdala and frontal cortex. The changes becom e m ore apparent with extension of heroin dependence tim e.