Objective:To analyze the specific immunoglobulin G (IgG) antibodies level in the population after the coronavirus disease 2019 (COVID-19) pandemic in Henan Province.Methods:A total of 5 178 peripheral venous blood samples were collected from 10 districts (counties) in Henan Province according to the national seroepidemiological survey program for COVID-19, and the method of cluster random sampling was adopted from March 6 to 15, 2023. Descriptive analysis was used for the basic data, history of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccination, SARS-CoV-2 infection of the respondents. The specific IgG antibody of SARS-CoV-2 was detected using chemiluminescence method. Statistical analysis was performed by using rank sum test, Kruskal Wallis test, and Dunn′s test.Results:The overall positive rate of SARS-CoV-2-specific IgG antibody was 83.35%(4 316/5 178). There were statistically significant differences in the specific IgG antibodies against SARS-CoV-2 produced by people of different sexes, different ages, infected or not, vaccinated or not, and vaccinated with different doses of SARS-CoV-2 vaccine ( Z=3.60, H=195.32, Z=6.10, 18.08, H=382.70, respectively, all P<0.001). The specific IgG antibodies produced by unvaccinated+ uninfected group, unvaccinated+ infected group, vaccinated+ uninfected group, and vaccinated+ infected group were 3.54(0.98, 11.00), 60.65(2.33, 84.80), 133.00(59.80, 173.00), and 142.00(98.30, 176.00), respectively. And the difference was statistically significant( H=354.62, P<0.001). The specific IgG antibodies of uninfected people increased with the increase of inoculum times( H=287.00 and 98.48, both P<0.001). The specific IgG antibodies of people who were not infected with SARS-CoV-2 in the groups of whose interval from the last inoculation of SARS-CoV-2 vaccine to blood collection was less than three months, three to six months and more than six months were 171.86(156.04, 196.57), 71.71(17.08, 110.38) and 132.14(57.59, 172.25), respectively, and the difference was statistically significant ( H=19.93, P<0.001). Among them, the absolute difference between the less than three months group and the three to six months group was statistically significant ( Z=3.67, P<0.001), and the absolute difference between the less than three months group and the more than six months group was statistically significant ( Z=3.47, P<0.001). The specific IgG antibodies level in the less than three months group was the highest. Conclusions:There is a certain correlation between the number of SARS-CoV-2 vaccine doses and the specific IgG antibodies level in uninfected people. The specific IgG antibodies could maintain a high level for three months after immunization.

Objective:To analyze the genetic evolution and molecular characteristics of H7N9 avian influenza virus (AIV) isolated in a live poultry market.Methods:Samples such as poultry feces, sewage, and hair removal machine and chopping board swabs were collected. Real-time fluorescent quantitative PCR was used to detect influenza A virus and H7N9 AIV in the samples. The whole genome of H7N9 AIV was amplified with influenza A virus universal primers and sequenced. BLAST and MEGA X were used for sequence alignment, phylogenetic analysis and molecular characterization.Results:Seven poultry-related environment samples were collected in the live poultry market in Xuchang city in February 2023, and four were positive for H7N9 AIV. The whole genome sequences of three H7N9 AIV isolates were successfully obtained, and the isolates shared high nucleotide identity in different genes (98.37%-100.00%). BLAST analysis showed they were highly identical to H7N9 strains isolated from domestic poultry in China from 2020 to 2021. Genetic evolution analysis showed that the three isolates clustered in the same branch and were closer to the recent environmental isolates than to the recent strains isolated from human or avian. Through comparison with the sequences of the representative strains in different periods, it was found that the isolated strains in this study showed high avian pathogenicity with four amino acids KRAA inserted at the cleavage site; the hemagglutinin receptor-binding site was QSG, which was an avian binding receptor; there was a G186I mutation in hemagglutinin. Mammalian-adaptive mutation E627K was not detected in polymerase basic protein 2. Mutations (R292K and I38T) associated with drug resistance to neuraminidase inhibitor (oseltamivir) and polymerase acidic protein inhibitor (baloshavir) were not detected, suggesting that these isolates remained susceptible to these drugs. A S31N mutation was found in M2 protein, indicating they were resistant to alkamines.Conclusions:The three H7N9 AIV strains isolated in the live poultry market have high avian pathogenicity, but there are no significant increase in mutations related to the binding ability to human receptors, mammalian pathogenicity, viral transmissibility, or drug resistance as compared with previous representative strains causing human or avian infection.

Objective:To analyze the reinfection rates in people previously infected with SARS-CoV-2 in Zhengzhou and Yuzhou cities (first infected with Delta/B.1.617.2 variant), and Anyang city (first infected with Omicron/BA.1.1 variant) in January 2022 and the population characteristics, and compare the differences in antibody levels among different populations.Methods:Serum samples were collected from 371 previously infected, 134 reinfected and 19 uninfected people for IgG antibody detection. Among them, serum samples from 45 previously infected, 44 reinfected and 19 uninfected people were tested with different novel coronavirus variants (early original strain, BA.5.2 variant, XBB.1.5 variant) for neutralizing antibody detection.Results:The rate of reinfection was 32.82% (85/259) in Zhengzhou and Yuzhou cities, and 19.92% (49/246) in Anyang city. The IgG antibody level in reinfected people was higher than that in previously infected and uninfected people ( P<0.05). The IgG antibody level in uninfected group was higher in people vaccinated within three months than in those vaccinated six months ago ( P<0.05). The IgG antibody level in the group receiving four doses of vaccine was higher than that in the group receiving three doses of vaccine ( P<0.05). The results of true virus neutralization antibody detection showed that in the Zhengzhou and Yuzhou cases, the level of neutralization antibody against the early original strain was higher than those against the BA.5.2 variant and the XBB.1.5 variant ( P<0.05), and the level of neutralizing antibody against BA.5.2 variant was higher than that against XBB.1.5 variant ( P<0.05). In Anyang city cases, the level of neutralizing antibody against the early original strain was higher than those against BA.5.2 variant and XBB.1.5 variant ( P<0.05); in the reinfected population, the level of neutralizing antibody against the early original strain was higher than that against the XBB.1.5 variant ( P<0.05). In addition, the levels of all neutralizing antibodies in both previously infected and reinfected people were higher than those in uninfected people ( P<0.05). The level of neutralizing antibody in the infected population in Zhengzhou and Yuzhou cities was higher than that in the infected population in Anyang city and in uninfected population ( P<0.05). The levels of antibodies against BA.5.2 and anti-XBB.1.5 variants in infected people in Zhengzhou and Yuzhou cities were higher than those in uninfected people ( P<0.05). The level of neutralizing antibody against BA.5.2 variants in the previously infected population in Anyang city was higher than that in the uninfected population ( P<0.05), and the level of neutralizing antibody against XBB.1.5 variants in the infected population in Anyang city was higher than that in the uninfected population ( P<0.05). Conclusions:After infection with SARS-CoV-2, the neutralizing antibodies produced in the human body have a certain cross-protection effect on other variants, but the antibody level will gradually decrease over time. Protection from a previous early SARS-CoV-2 variants infection against the current main circulating Omicron variants (such as XBB variants) is low, and the immunity conferred by pervious infection or booster vaccination may not be able to provide sufficient protection against new variants.

Objective:To analyze the whole genome of Omicron variants causing the first local Omicron outbreak in Henan Province and to investigate the mutations in the SARS-CoV-2 genome for source tracing.Methods:Respiratory tract samples from COVID-19 cases in the Omicron outbreak in Henan Province from January 7 to 29, 2022 were subjected to whole-genome sequencing and sequence alignment analysis. Whole-genome identity, variations and evolution of the Omicron variants were analyzed.Results:Through high-throughput sequencing, the whole-genome sequences of SARS-CoV-2 were obtained from 120 cases, which accounted for 25.64% (120/468) of all COVID-19 cases in Anyang during the same period. Compared with the genome of Wuhan reference strain (NC_045512.2), there were 57-59 nucleotide mutation sites in the 120 whole genome sequences, and one or two nucleotide mutation sites were added to the shared 57 nucleotide sites. All of the 120 strains were VOC/Omicron (BA.1.1) variants and shared high homology. The whole-genome sequence obtained from the first case A contained 57 nucleotide mutation sites, while apart from the 57 identical nucleotide mutation sites, one specific mutation site (C1594T) was found in the whole-genome sequence obtained from the first case B, suggesting that the two cases were in the same transmission chain. After comparing with the database of domestic and imported cases by the Chinese Center for Disease Control and Prevention and the Henan Provincial Center for Disease Control and Prevention, it was found that the current outbreak was linked with the same transmission chain as the existing local epidemics in other provinces. Moreover, epidemiological investigation showed that on January 2, case A had come into contact with her cousin and his family who returned from an affected area outside the province.Conclusions:Based on the gene sequencing results and epidemiological investigation, the COVID-19 outbreak in Anyang city, Henan Province was a local epidemic and the source of it was a college student who returned to Anyang city from other province on December 28, 2021. These infections were linked to the same transmission chain as the existing local infection in other provinces.

Objective To observe the impacts of acupuncture and moxibustion on kelch like epichlorohydrin related protein 1(Keap1)and nuclear factor E2 related factor 2(Nrf2)signal pathway in kidney tissue of cisplatin(DDP)-induced kidney injury model mice,and to explain the protective mechanism of acupuncture and moxibustion on improving kidney injury caused by DDP.Methods Forty SPF male KM rats were randomly divided into 4 groups,with 10 rats in each group.One day before the start of treatment,the three groups of mice outside the blank group were intraperitoneally injected with cisplatin 10 mg·kg-1 according to their body weight,and the blank group was injected with the same dose of 0.9%NaCl solution.The model was established 24 hours later.Both acupuncture group and moxibustion group selected"Dazhui"(GV14),"Ganshu"(BL18),"Shenshu"(BL23)and"Zusanli(Housanli)"(ST36)for acupuncture and moxibustion respectively,once a day for 5 days.The other two groups were fixed every day without treatment.After fasting for 1 day,the contents of BUN,Scr,CysC and NGAL in serum and Keap1 and Nrf2 in renal tissue were detected by ELISA;Western blot and Real-time PCR were used to detect the protein expression and gene transcription of Keap1 and Nrf2 in the kidney tissue of mice in each group.Results Compared with the blank group,the content of Keap1 protein,protein expression and relative expression of mRNA in the model group increased(P<0.05),the content of Nrf2 protein,protein expression decreased(P<0.05),Nrf2 relative expression of mRNA increased(P<0.05);Compared with the model group,the content of Keap1 protein,the expression of Keap1 protein and the relative expression of Keap1 mRNA in the kidney of mice in the acupuncture and moxibustion groups decreased(P<0.05);Nrf2 protein content,protein expression and relative mRNA expression increased(P<0.05).Conclusion Acupuncture and moxibustion can improve the renal function of DDP renal injury model mice and enhance their antioxidant stress ability,so as to improve the renal injury caused by DDP chemotherapy.Its mechanism may be related to Keap1/Nrf2 signal pathway.

Objective:To analyze the genome characteristics and variations in nucleotides and amino acids of SARS-CoV-2 causing an outbreak in Henan Province in November 2021 and perform the traceability analysis.Methods:In this study, throat swab specimens from cases in the acute phase were collected and tested for the nucleic acids of SARS-CoV-2 by real-time fluorescent RT-PCR. SARS-CoV-2 nucleic acid-positive samples were subjected to high-throughput genome sequencing and whole-genome alignment analysis.Results:The median Ct values of ORF1ab gene and N gene in 70 positive specimens was 26.41 (15.58 to 39.27) and 24.43 (12.04 to 39.74), respectively. Compared with the sequence of Wuhan-Hu(NC_045512) reference strain, 47 to 49 nucleotide mutations sharing 47 nucleotide mutation and 41 amino acid mutations were found in 63 strains of successfully sequenced SARS-CoV-2. Nine nucleotide mutations and 12 amino acid mutations were found in the spike protein. The index case shared 47 mutations with the Russian imported cases in Henan Province on October 14 and the local cases in Jiangxi Province in October. Moreover, their genomes were highly homologous and they all belonged to the Delta variant (AY.122 evolutionary branch).Conclusions:Continuous monitoring of imported COVID-19 cases and prolonging the period of quarantine were needed to reduce the risk of local outbreak and epidemic caused by imported COVID-19 cases. Analysis of the genomic characteristics of SARS-CoV-2 and the variations in nucleotides and amino acids was conducive to trace the origin of COVID-19 outbreak quickly and provide reference for precise control.

OBJECTIVE: To construct an adenovirus vector expressing artificial splicing factor capable of regulating alternative splicing of Yap1 in cardiomyocytes. METHODS: The splicing factors with different sequences were constructed against Exon6 of YAP1 based on the sequence specificity of Pumilio1. The PCR fragment of the artificially synthesized PUF-SR or wild-type PUFSR was cloned into pAd-Track plasmid, and the recombinant plasmids were transformed into E. coli DH5α for plasmid amplification. The amplified plasmids were digested with Pac I and transfected into 293A cells for packaging to obtain the adenovirus vectors. Cultured neonatal rat cardiomyocytes were transfected with the adenoviral vectors, and alternative splicing of YAP1 was detected using quantitative and semi-quantitative PCR; Western blotting was performed to detect the signal of the fusion protein Flag. RESULTS: The transfection efficiency of the adenovirus vectors was close to 100% in rat cardiomyocytes, and no fluorescent protein was detected in the cells with plasmid transfection. The results of Western blotting showed that both the negative control and Flag-SR-NLS-PUF targeting the YAPExon6XULIE sequence were capable of detecting the expression of the protein fused to Flag. The results of reverse transcription-PCR and PCR demonstrated that the artificial splicing factor constructed based on the 4th target sequence of YAP1 effectively regulated the splicing of YAP1 Exon6 in the cardiomyocytes (P < 0.05). CONCLUSION We successfully constructed adenovirus vectors capable of regulating YAP1 alternative splicing rat cardiomyocytes.
Adenoviridae/metabolism* ; Alternative Splicing ; Animals ; Animals, Newborn ; Escherichia coli/metabolism* ; Genetic Vectors ; Myocytes, Cardiac/metabolism* ; Plasmids ; RNA Splicing Factors/metabolism* ; Rats ; Transfection

Objective:To investigate the effect of hemoglobin on Shp2 expression in brain tissues and adherens junction in blood brain barrier in rats after cerebral hemorrhage.Methods:Eighty male SD rats were randomly allocated into sham-operated group and groups of cerebral hemorrhage for 6 h, 24 h, 3 d, and 7 d ( n=16). Rat models in the groups of cerebral hemorrhage were established by intracerebral injection of 20 μL of hemoglobin. Six h, 24 h, 3 d, and 7 d after injection, neurological functions were assessed by Longa scale; brain water content and brain organ coefficient in the harvested cerebral tissues were calculated by wet-dry weighting method; Shp2 mRNA expression was analyzed by real-time fluorescence quantification reverse transcription PCR (qRT-PCR); Shp2 positive neurons was detected by immunohistochemistry; the protein expressions of Shp2, α-catenin, β-catenin and vascular endothelial (VE)-cadherin, and phosphorylated (p-) α-catenin, p-β-catenin and p-VE-cadherin were explored by Western blotting. Results:As compared with those in the sham-operated group, Longa scale scores in groups of intracerebral hemorrhage for 6 h, 24 h, 3 d and 7 d were significantly increased ( P<0.05). As compared with the sham-operated group, groups of intracerebral hemorrhage for 24 h, 3 d and 7 d had significantly increased brain water content and brain organ coefficient, statistically reduced Shp2 mRNA expression, statistically samller amount of Shp2 positive neurons, and statistically reduced Shp2 protein expression, and significantly increased protein expressions of p-α-catenin, p-β-catenin and p-VE-cadherin ( P<0.05). Conclusion:Intracerebral injection of hemoglobin can downregulate Shp2 expression and promote phosphorylation of adherens junction proteins, therefore induce the disruption of adherens junction, which might be a critical mechanism of blood-brain barrier disruption and brain edema.

Objective: To explore the flap design and clinical applications of the propeller flaps based on perforators from different branches of the lateral circumflex femoral artery in defect reconstruction. Methods: Between September 2009 and December 2018, 27 patients with soft tissue defects from lower extremities were involved in this study, including 15 males and 12 females, with an average age of 34.6 years old (range, 3 to 73 years old). Before surgery, the type of the flap to be used was designed preliminarily by evaluating the location, size and shape of the lesion or defect. The perforators of the lateral circumflex femoral artery were explored using an ultrasound Doppler probe, marked on skin. The computed tomographic angiography was also used to get more information of the branches of the lateral circumflex femoral artery when possible. The propeller flaps were divided into typeⅠ, Ⅱ, Ⅲ, and Ⅳ according to the perforators that originated from the transverse, descending, oblique, and rectus femoris branches of the lateral circumflex femoral artery, respectively. The type Ⅱ flap was subdivided into type Ⅱa and type Ⅱb flaps that were based on antegrade and reverse flow from the descending branch. The defects were reconstructed using the perforator propeller flap. Results: Twenty-seven patients underwent reconstruction of defects using the propeller flaps based on perforators from different branches of the lateral circumflex femoral artery including 3 type Ⅰ flap, 12 type Ⅱa, 2 type Ⅱb, 10 type Ⅲ and 3 type Ⅳ. The size, pedicle length and rotation angle of the flaps were 12 cm×6 cm to 30 cm×15 cm, 4 cm to 15 cm, and 60 to 180 degrees, respectively. Total necrosis occurred in one flap and small-sized distal necrosis in another one. Minor complications occurred in two flaps and the remaining propeller flaps survived completely. All patients were followed up from one to 25 months and mean follow-up time was 9.9 months. Tumor recurrence was noticed in one patient. All patients were satisfied with the final functional and aesthetic outcomes. Conclusions For appropriate cases, reconstruction of defects from the groin to the knee could be achieved by using the propeller flaps based on perforators from different branches of the lateral circumflex femoral artery, with advantages of easy-to-operate and minor donor-site morbidity.

OBJECTIVE: To explore the feasibility of rapid and accurate three-dimensional (3D) image reconstruction using Uromedix-3D software for urological surgery. METHODS: The original renal thin-slice enhancement CT data were obtained from patients with kidney lesions treated in our hospital between December, 2015 and October, 2018. The self-developed Uromedix- 3D system was used to reconstruct the normal kidney structures, blood vessels, collecting systems and the lesions. The spatial anatomic relationships of the structures were measured and digitized for surgical planning. RESULTS: 3D reconstruction of the kidneys was performed in a total of 173 cases, and the mean time for reconstruction was 31.24±2.012 min. Of these cases, 147 (84.9%) had renal tumors, and 2 had renal tumors with tumor thrombus. In addition to renal tumors, the Uromedix-3D system was also used for reconstructing other lesions including UPJO, kidney stones and retroperitoneal masses. Renal artery reconstruction was performed in 170 cases, which allowed observation of the precise terminal branches (up to 7th grade arterial branch) of the artery; 109 (64%) cases showed the 5th grade arterial branch or above. Renal artery variations were detected in 37 cases, including accessory renal artery (24 cases) and multiple renal arteries (13 cases). The renal veins were reconstructed in 164 cases, and second grade or above (up to the 4th grade) vein branches were observed in 138 (84%) cases. CONCLUSIONS Uromedix-3D system can accurately and efficiently reconstruct the 3D structure of human kidneys and the renal lesions based on enhanced CT data. The reconstructed 3D model allows objective assessment of the spatial anatomical relationship of the lesions to provide assistance in surgical planning.
Humans ; Imaging, Three-Dimensional ; Kidney ; Kidney Calculi ; diagnostic imaging ; surgery ; Kidney Neoplasms ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed