Objective:To analyze phylogenetic structure and molecular characteristics of H5N6 avian influenza virus (AIVs) isolated from live poultry market (LPM).Methods:Oropharyngeal and cloacal swabs from poultry, and environmental samples were collected from LPM in Urumqi in December 2018, AIVs were isolated and identified by inoculation of chicken embryo, hemagglutination test and RT-PCR, the viral whole genome was amplified with the universal primers of influenza A virus, and then sequenced, pairwise sequence alignments, phylogenetic and molecular characteristics analysis were performed by BLAST, Clustal W, MEGA-X and DNAStar software.Results:Six strains of H5N6 AIVs were isolated from poultry samples, the identity between the viral genes was high (99.4%-100.0%), so the isolates were the same source. BLAST analysis revealed that the viral NP sequence had the highest identity (99.7%) with H5N6 AIVs isolated from poultry in Suzhou, while the sequence of the remaining 7 viral genes had the highest identity (99.0%-100.0%) with H5N6 AIVs isolated from environment in Guangdong during 2017 to 2018. Phylogenetic analysis showed that the viral HA belonged to Clade 2.3.4.4C, and the viral HA, NA, PB1, PA, NP, and MP were all clustered together with H5N6 AIVs isolated from mink in Eastern China in 2018, while the PB2 and NS were clustered together with H5N6 AIVs isolated from environment in Guangdong from 2017 to 2018. The HA cleavage site contained multiple basic amino acid residues, which was highly pathogenic AIVs (HPAIVs). S137A and T160A mutations of HA could increase binding to human-type receptor SAα2, 6-Gal. Additionally, the viral multiple mutations, including 59-69 deletion in NA, the L89V, G309D, R477G, I495V, I504V, D391E, and A661E in PB2, as well as the P42S, D92E, and 80-84 deletion in NS1, could enhance the viral virulence and pathogenicity to mammals. Conclusions:The 6 strains of H5N6 HPAIVs isolated from LPM have relatively close genetic relationship with H5N6 AIVs isolated from mink in Eastern China and environment in Guangdong during 2017 to 2018, the viral multiple mutations could increase its pathogenicity to mammals, which could pose a potential risk to public health.

Objective To measure histidine triad nucleotide?binding protein 1(HINT1)protein expression and gene promoter methylation, and to analyze the relationship between HINT1 gene promoter methylation and clinical pathological features of melanoma. Methods Fifty?six patients with melanoma and 51 patients with nevus were enrolled as subjects and controls, respectively. Methylation?specific PCR (MSP) was performed to measure the methylation of HINT1 gene promoter in lesional and paratumoral tissue specimens from the patients with melanoma, as well as in lesional specimens from the patients with nevus. Immunohistochemistry was carried out to quantify the expression of HINT1 protein in these tissue specimens. Results MSP showed that the methylation rate of HINT1 gene promoter was significantly higher in melanoma tissues than in paratumoral and nevus tissues(76.8%[43/56]vs. 33.9%[19/56]and 35.3%[18/51], χ2 = 20.810 and 18.749, respectively, both P < 0.05), but was insignificantly different between paratumoral and nevus tissues(χ2=0.022, P>0.05). Immunohistochemistry revealed that the expression rate of HINT1 was 21.4%(12/56)in melanoma tissues, compared to 82.4%(42/51)in nevus tissues(χ2 = 39.633, P <0.01). There was a significant difference in the methylation rate of HINT1 promoter between HINT1?positive and ?negative melanoma tissues(6/12 vs. 37/44[84.1%], P<0.05), and between Clark levelⅠ-ⅡandⅢ-Ⅴmelanoma tissues(59.1%[13/22]vs. 88.2%[30/34],χ2=6.365,P=0.012). Conclusions HINT1 protein is lowly expressed in melanoma, which may be associated with high methylation of its gene promoter. Moreover, the high methylation ofHINT1 gene promoter may be involved in the initiation and progression of melanoma.

Objective To investigate the expression of regulators of G protein signaling(RGS), including RGS2, RGS3 and RGS4 in OLETF rats, as well as the effects of metformin on these expressions. Methods LETO rats were used as control group. Eight-week-old male OLETF rats were assigned to two guoups randomly:model and trial(metfomin dose during 8~(th) to 22~(nd) weeks:300mg kg~(-1)·d~(-1);during 23rd to 28th weeks:400 mg·kg~(-1) ·d~(-1))groups. Expressions of RGS mRNA in aorta and heart werequantified by real-time PCR. Results RGS2, RGS3 and RGS4 mRNA of the thoracic aorta and left ventricle were significantly higher in model group than in control group (P<0.01). Compared with model group, metformin significantly reduced their mRNA in trial group (P<0.01). Conclusions Upregulation of RGS2, RGS3 and RGS4 mRNA expression in the thoracic aorta and left ventricle of OLETF rats is in correlation with cardiovascular lesions; while downregulation of their expression is in correlation with the action of metformin.

Objective:To investigate the effect of edaravone, a free radical scavenger, on the regulation of retinal autophagy and the protection of photoreceptor cells at the early stage of experimental retinal detachment (RD) in rats.Methods:Fifty-one adult male Sprague-Dawley rats were used for RD model establishment, and another 24 rats were served as PBS injection group.The RD model was established via subretinal injection of 0.5% sodium hyaluronate into the right eye of the rats and the rats with successful modeling were randomly divided into RD model group and edaravone treatment group.The rats in the edaravone treatment group were given edaravone of 3 mg/kg intraperitoneally, twice a day after modeling, and the rats in the PBS injection group and RD model group were given equal volume of normal saline.Rats were sacrificed on the 1st day, 3rd day and 7th day following modeling.The T-superoxide dismutase (T-SOD) activity and malondialdehyde (MDA) content in the intraocular fluid was detected.The expression levels of superoxide dismutase 2 (SOD2), nuclear factor E2-related factor 2 (Nrf2), autophagy related gene 4 (Atg4), microtubule-associated protein 1 light chain 3B (LC3B) and other proteins in retinal tissue were identified by Western blot analysis.TUNEL staining was performed on paraffin sections of the whole eyeball to analyze the apoptosis of photoreceptor cells.The study protocol was approved by an Ethics Committee of Xi'an Fourth Hospital (No. 2016016). The use and care of animals complied with the Regulations on the Administration of Experimental Animals.Results:The RD area was more than 60% in rat eyes of RD model.There were significant differences in MDA content and T-SOD activity among different groups at various time points (MDA: Fgroup=385.513, P<0.01; Ftime=13.021, P<0.01.T-SOD: Fgroup=48.865, P<0.01; Ftime=7.700, P=0.003). Compared with the PBS injection group, the MDA concentration was significantly increased and the T-SOD activity was significantly decreased in the RD group and edaravone treatment group on the 1st, 3rd and 7th day after modeling (all at P<0.05). The MDA concentration was significantly reduced and the T-SOD activity was significantly elevated in the edaravone treatment group on the 1st, 3rd and 7th day after modeling in comparison with those of the RD group (all at P<0.05). Compared with the PBS injection group, the relative expression levels of SOD2 and Nrf2 proteins were significantly increased in the RD group and edaravone treatment group on the 1st, 3rd and 7th day after modeling (all at P<0.05), and Atg4 and LC3B-Ⅱ/LC3B-Ⅰ were significantly increased on the 1st, 3rd and 7th day after modeling (all at P<0.05). The expression level of SOD2 in the edaravone treatment group was significantly higher than that in the RD group on the 1st, 3rd and 7th day after modeling (all at P<0.05), and the expression level of Nrf2 was significantly increased in the edaravone treatment group on the 1st and 3rd day after modeling compared with that of the RD group (both at P<0.05), and the expression levels of Atg4 and LC3B-Ⅱ/LC3B-Ⅰ were significantly increased in the edaravone treatment group on the 3rd day after modeling in comparison with those of the RD group (both at P<0.05). No significant TUNEL positive cells were observed in PBS injection group at all time points, and TUNEL positive cells were observed on the 1st, 3rd and 7th day after modeling in the RD group, and the expression level of caspase-3 in the RD group was significantly increased in comparison with that of the PBS injection group ( P<0.05). The apoptosis of photoreceptor cells and the expression level of caspase-3 in edaravone treatment group were significantly decreased in comparison with those of the RD group on the 1st, 3rd and 7th day after modeling (all at P<0.05). Conclusions:The intraperitoneal injection of edaravone, twice a day, can significantly improve the antioxidant capacity of the retina after experimental RD in rats, regulate retinal autophagy and reduce the apoptosis of photoreceptor cells in early-stage RD.

Objective To construct and preliminarily apply a graded exercise rehabilitation program for patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD),and to provide a theoretical basis for the scientific implementation of exercise rehabilitation by medical staff.Methods Based on Triangle model and evidence-based method,the first draft of the graded exercise rehabilitation program was constructed,and the items were revised through 2 rounds of expert consultations from March to October,2023.The graded exercise rehabilitation program was preliminarily applied in 10 patients with AECOPD.Results The effective recovery rates of the 2 rounds of expert consultation questionnaires were 100％;the expert authority coefficients were 0.857 and 0.863;the coefficients of variation were 0-0.285 and 0.052-0.244;the Kendall's harmony coefficients were 0.167 and 0.145,respectively(all P＜0.001).The final plan includes 4 first-level indicators,13 second-level indicators,and 25 third-level indicators.The scores of 6 min walking test,mMRC and CAT after exercise were improved compared with those before exercise,and the differences were statistically significant(all P＜0.05).Conclusion The graded exercise rehabilitation program for patients with AECOPD constructed in this study has good scientificity and practicability,which can provide references for clinical implementation of exercise rehabilitation.

Objective To compare the effect of storage autologous blood component transfusion versus storage autologous whole blood transfusion on the cellular immune function and hemorheology in the patients undergoing spinal surgery.Methods Forty patients of both sexes,aged 32-60 yr,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,undergoing elective multilevel spinal surgery,were divided into 2 groups (n =20 each) using a random number table:stored autologous whole blood transfusion group (group A) and stored autologous blood component transfusion group (group B).Before blood sampling (T0),immediately after blood sampling (T1) and at the end of surgery (T2),arterial blood samples were collected for determination of red blood cell count (RBC),hemoglobin (Hb),hematocrit (Hct),erythrocyte aggregation index (EAI) and erythrocyte rigidity index (ERI).Venous blood samples were collected at T0,T2 and on day 6 after surgery (T3),the distribution of T lymphocyte subsets (percentage of CD3+,CD4+ and NK cells) was measured,and CD4+/CD8+ ratio was calculated.Results Compared with the baseline at T0,the percentage of CD3+,CD4+ and NK cells and CD4+/CD8+ ratio were significantly decreased at T2,3 in group A and at T2 in group B,and RBC,Hb and Hct were significantly decreased at T1,and EAI and ERI were decreased at T1,2 in two groups (P＜0.05).Compared with group A,the percentage of CD3+,CD4+ and NK cells and CD4+/CD8+ ratio were significantly increased at T3 (P＜0.05),and no significant change was found in RBC,Hb,Hct,EAI or ERI at each time point in group B (P＞0.05).Conclusion The effect of storage autologous blood component transfusion on cellular immune function is mitigated than that of storage autologous blood transfusion and the effects on hemorheology are comparable in the patients undergoing spinal surgery.

Background: The H5 avian influenza viruses (AIVs) of clade 2.3.4.4 circulate in wild and domestic birds worldwide. In 2017, nine strains of H5N6 AIVs were isolated from aquatic poultry in Xinjiang, Northwest China. Objectives: This study aimed to analyze the origin, reassortment, and mutations of the AIV isolates. Methods: AIVs were isolated from oropharyngeal and cloacal swabs of poultry. Identification was accomplished by inoculating isolates into embryonated chicken eggs and performing hemagglutination tests and reverse transcription polymerase chain reaction (RT-PCR). The viral genomes were amplified with RT-PCR and then sequenced. The sequence alignment, phylogenetic, and molecular characteristic analyses were performed by using bioinformatic software. Results: Nine isolates originated from the same ancestor. The viral HA gene belonged to clade 2.3.4.4B, while the NA gene had a close phylogenetic relationship with the 2.3.4.4C H5N6 highly pathogenic avian influenza viruses (HPAIVs) isolated from shoveler ducks in Ningxia in 2015. The NP gene was grouped into an independent subcluster within the 2.3.4.4B H5N8 AIVs, and the remaining six genes all had close phylogenetic relationships with the 2.3.4.4B H5N8 HPAIVs isolated from the wild birds in China, Egypt, Uganda, Cameroon, and India in 2016–2017, Multiple basic amino acid residues associated with HPAIVs were located adjacent to the cleavage site of the HA protein. The nine isolates comprised reassortant 2.3.4.4B HPAIVs originating from 2.3.4.4B H5N8 and 2.3.4.4C H5N6 viruses in wild birds. Conclusions These results suggest that the Northern Tianshan Mountain wetlands in Xinjiang may have a key role in AIVs disseminating from Central China to the Eurasian continent and East African.

Background: The H5 avian influenza viruses (AIVs) of clade 2.3.4.4 circulate in wild and domestic birds worldwide. In 2017, nine strains of H5N6 AIVs were isolated from aquatic poultry in Xinjiang, Northwest China. Objectives: This study aimed to analyze the origin, reassortment, and mutations of the AIV isolates. Methods: AIVs were isolated from oropharyngeal and cloacal swabs of poultry. Identification was accomplished by inoculating isolates into embryonated chicken eggs and performing hemagglutination tests and reverse transcription polymerase chain reaction (RT-PCR). The viral genomes were amplified with RT-PCR and then sequenced. The sequence alignment, phylogenetic, and molecular characteristic analyses were performed by using bioinformatic software. Results: Nine isolates originated from the same ancestor. The viral HA gene belonged to clade 2.3.4.4B, while the NA gene had a close phylogenetic relationship with the 2.3.4.4C H5N6 highly pathogenic avian influenza viruses (HPAIVs) isolated from shoveler ducks in Ningxia in 2015. The NP gene was grouped into an independent subcluster within the 2.3.4.4B H5N8 AIVs, and the remaining six genes all had close phylogenetic relationships with the 2.3.4.4B H5N8 HPAIVs isolated from the wild birds in China, Egypt, Uganda, Cameroon, and India in 2016–2017, Multiple basic amino acid residues associated with HPAIVs were located adjacent to the cleavage site of the HA protein. The nine isolates comprised reassortant 2.3.4.4B HPAIVs originating from 2.3.4.4B H5N8 and 2.3.4.4C H5N6 viruses in wild birds. Conclusions These results suggest that the Northern Tianshan Mountain wetlands in Xinjiang may have a key role in AIVs disseminating from Central China to the Eurasian continent and East African.

Chronic pulmonary aspergillosis (CPA) is a serious fungal infection, which complicates pulmonary tuberculosis, chronic obstructive pulmonary disease and sarcoidosis with high mortality. At present, the treatment of CPA include drug treatment and surgical treatment. The treatment plan should be determined by the type, clinical manifestations and surgical indications of the patients. Nevertheless, there is no standard plan, and more research is needed to improve it. Combined with the current literature reports, this article reviews the current and progress in the treatment of CPA.

Objective:To analyze the changing trend of intracerebral hemorrhage (ICH) incidence among residents with different characteristics during 9 years of comprehensive hypertension prevention and control (hypertension prevention and control) in Tengzhou from 2013 to 2021.Methods:From January 1, 2013 to December 31, 2021, the new ICH cases collected by the Center for Disease Control and Prevention in Tengzhou City were analyzed to calculate the incidence of ICH, and the trend of its distribution was analyzed among residents with different ages, sexes, and between urban and rural areas. The registered population information came from Tengzhou Public Security Bureau. Age and sex standardized incidence was calculated based on the 7th National Population Census in 2020. The Cochran-Armitage test was used to analyze the time and age trends of the incidence.Results:The overall ICH crude and standardized incidence in Tengzhou City decreased from 97.30/100 000 to 52.13/100 000 ( Z=-9.93, P<0.001) and 119.30/100 000 to 50.69/100 000 ( Z=-15.40, P<0.001) from 2013 to 2021, and both elevated to form a single peak in 2020, with 22.58% ( χ 2=24.02, P<0.001) and 18.09% ( χ 2=17.08, P<0.001) higher than in 2019, respectively. The trends in male and female incidence over the same period were similar to the overall trends, and the incidence was higher in males than in females in all years. The incidence of ICH increased with age in all years. The difference of increase in male incidence rate in 2020 was statistically significant in three age groups ≥45 years compared with 2019 (36.29%, 23.57% and 16.18%, respectively, χ 2=6.73, 4.65, 4.00, P<0.001). The incidence of ICH decreased by 70.07% and 36.23% ( Z=18.44, 5.22, P<0.001) in urban and rural areas respectively from 2013 to 2021, whereas increased by 34.15% ( χ 2=10.88, P<0.01) and 22.08% ( χ 2=18.63, P<0.001) in 2020 compared with 2019 separately. Conclusions:The incidence of ICH in Tengzhou from 2013 to 2021 showed a significant downward trend, with the decrease in the incidence of ICH in women exceeding that in men. The decrease in the incidence of ICH in urban areas exceeded that in rural areas, and male morbidity seemingly had a younger trend.