Abstract: Objective To analyze the genetic evolution and molecular characteristics of the H5N6 avian influenza virus A/Changsha/1/2022(H5N6) in a human infection case in Changsha City, providing insights for the prevention and control of H5N6 avian influenza transmission to humans. Methods The sample underwent whole-genome sequencing on a third-generation sequencing platform. Reference sequences were downloaded from the NCBI and GISAID databases for comparison. The phylogenetic tree was constructed and key amino acid mutation sites were analyzed using MEGA7 software. Results The sequence analysis results revealed that the strain studied belongs to the H5 subtype Clade 2.3.4.4b branch. The PB1 gene shared 99.69% homology with the H5N6 virus A/duck/Hunan/S40199/2021(H5N6) from Hunan Province; PB2 had 98.58% homology with A/Whooper swan/Sanmenxia/Y36/2020(H5N8); and other sequences showed high homology with the strain A/Guangdong/1/2021(H5N6) discovered in Guangzhou in 2021. These results indicate that an infection with a recombinant H5N6 subtype avian influenza virus caused the case. The amino acid composition of the cleavage site was RERRRKR↓GLF, consistent with high-pathogenicity avian influenza characteristics. The HA sequence showed no mutations at the Q226L and G228S sites, indicating the preservation of characteristics binding to avian receptors. However, mutations at the S127P, S137A, T160A, T192R, and A267T sites increased the virus's affinity for humans. A deletion found in positions 59-70 of the NA protein stalk implies enhanced viral virulence in mammals. Mutations were noted in the internal genes; S622G in PB1, K389R and V598T in PB2, N409S in PA, N30D and T215A in M1, and P42S in NS1, which may enhance the virulence of avian influenza virus in infecting mice. Conclusions The H5N6 virus strain detected in a human case in Changsha City in this study is highly pathogenic, prone to bind to avian receptors, but numerous mutations at key amino acid sites facilitate infection in humans. Continuous monitoring and research on H5N6 avian influenza virus should be strengthened

Abstract: Objective To analysis the genetic evolution characteristics of hemagglutinin (HA) gene of influenza A(H1N1)pdm virus in Changsha City from 2016-2023, to understand the trend of the HA genetic evolution and the mutations of the amino acid. It provides a scientific basis for the prevention and control of influenza epidemics, as well as the screening of vaccines under the new situation. Methods The A(H1N1)pdm09 virus strains from Changsha City from 2016 to 2023 were isolated using SPF chicken embryos, and then the HA genes were sequenced by MiSeq of Illumina Inc. The homology of HA gene was analyzed by MegAlign of the DNASTAR, and the phylogenetic tree was constructed using the Neighbor Joining (NJ) method in the Molecular Evolutionary Genetics Analysis version 11 (MEGA11). Results The homology of the HA gene of A(H1N1)pdm virus in Changsha from 2016 to 2023 was between 94.8%-99.9%, with the HA gene homology decreasing annually. The homology between the isolated strains of A(H1N1)pdm09 in Changsha City from 2016 to 2023 and the WHO recommended vaccine strain ranged from 96.8% to 99.0%, indicating a relatively good match between the flu isolates and the recommended vaccine strain. The phylogenetic tree of the HA gene of the A(H1N1)pdm09 influenza virus in Changsha City showed that the HA gene evolved into several different branches within the 6B branch, and it had currently evolved to 6B.1A.5a.2a branch. Constant mutations had occurred at the amino acid sites of the four antigenic determinant clusters of HA protein. Currently, amino acid mutations had occurred at 15 antigenic sites within the four antigenic determinant clusters, and the newly emerged A186T antigen mutant site in the isolates from 2023 was worth recent notice. The receptor-binding sites are relatively conserved in loop 130, minor amino acid mutations occurred in loop 220, whether the amino acid mutation site in loop 190 is becoming more stable needs to be further monitored. Taking A/California/07/2009 (CY121680) as the reference strain, most of the A(H1N1) pdm09 isolates in Changsha was increased 162 NQTY glycosylation site and was decreased 276 NTTC glycosylation site, and the glycosylation mutations at these two sites have become more stable recently. Conclusions The HA genes of influenza A(H1N1)pdm virus in Changsha are constantly evolving and mutating, suggesting influenza surveillance should be strengthened continuously.

Objective To establish a polymerase chain reaction (PCR) method for detecting Brucella spp .and to evaluate its ap‐plication .Methods The PCR primer aiming at the outer membrane protein(omp‐2) coding gene were designed and the PCR method for detecting Brucella spp .was established .By using DNA of 19 kinds of common bacteria and Brucella spp .,the specificity and sensitivity were evaluated .Three strains of suspected Brucella spp .were performed the nucleic acid detection .Then ,3 suspected strains of Brucella spp .were amplified by the PCR method with omp‐2 gene ,and the results were compare to the those by the cul‐ture method .Results The established Brucella spp .PCR detection method could only detect the Brucella spp .positive strain ,the control bacterium DNA did not appear the target band ;the sensitivity was 100 copies/response;the PCR method detected omp‐2 gene band in 3 strains of suspected Brucella spp .,which was identified as Brucella spp .and was same to the results by the isolation culture method .Conclusion The established PCR method for detecting Brucella spp .is accurate and rapid compare with the isola‐tion and culture method ,which is suitable for the rapid detection of brucellosis epidemic situation .

Marked by the Third Plenary Session of the Eleventh Central Committee of the CPC, Shanghai health care started a journey of reform and opening up as with other sectors. This essay reviews various stages Shanghai health care experienced and the achievements for the last 30 years since reform and opening up, and analyzes the challenges Shanghai health care faces currently. On this basis, the essay summaries the experiences and lessons learned from Shanghai health care reform and development.

Under the general requirements of China's national health reform, the basic framework of Shanghai's health reform was constructed at five different aspects the long term vision, goal, strategies, measures, supportive reform activities. The general thought of Shanghai's health reform can be concluded into two aspects: first, establishing a basic healthcare system of covering both urban and rural residents universally; second, enhancing the foundation of enabling sustainable health development.

Objective:To study the effect of OSAHS on adult nasal-cycle.Method:The nasal cycle of 20 healthy adults,18 patients of snoring and 22 patients of OSAHS were examined with acoustic rhinometry,which was performed every 30 minutes in 7 hours.Result:The ratio of nasal cycle in healthy adults was(19/20,95.0%), in snoring patients was(15/18,83.0%),in patients of OSAHS was(15/22,68.2%). The mean alteration amplitude of nasal cycle in healthy adults was significantly larger than that in patients with OSAHS (P<0.05).The distribution of the healthy adults and patients with OSAHS between the typical cycle categories was significantly different(P<0.05).Conclusion:The characteristics of nasal-cycle in of OSAHS patients were different with healthy adults,which maybe owing to change of physical function of nasal cavity.

OBJECTIVE: To study the effect of OSAHS on adult nasal-cycle. METHOD: The nasal cycle of 20 healthy adults, 18 patients of snoring and 22 patients of OSAHS were examined with acoustic rhinometry, which was performed every 30 minutes in 7 hours. RESULT: The ratio of nasal cycle in healthy adults was (19/20, 95.0%), in snoring patients was (15/18, 83.0%), in patients of OSAHS was (15/22, 68.2%). The mean alteration amplitude of nasal cycle in healthy adults was significantly larger than that in patients with OSAHS (P < 0.05). The distribution of the healthy adults and patients with OSAHS between the typical cycle categories was significantly different (P < 0.05). CONCLUSION The characteristics of nasal-cycle in of OSAHS patients were different with healthy adults, which maybe owing to change of physical function of basal cavity.
Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Nasal Cavity ; physiopathology ; Nose ; physiopathology ; Rhinometry, Acoustic ; Sleep Apnea, Obstructive ; physiopathology ; Snoring ; physiopathology

OBJECTIVE: To investigate the anatomical structures of cranial nerves in the cerebellopontine angle region to offer anatomical data for clinical operation. METHOD: A total of 52 adult cadaveric heads fixed in 10% formalin were used for this study. After cutting cerebellum and meningeal between transverse and sigmoid sinus, simulate operating method of retrosigmoid approach to observe the cranial nerves. RESULT: External diameter and length of left V, VII, VIII, IX cranial nerves are (2.54 +/- 0.84) mm and (6.79 +/- 2.51) mm, (1.18 +/- 0.31) mm and (9.89 +/- 2.66) mm, (2.17 +/- 0.52) mm and (9.92 +/- 2.61) mm, (0.77 +/- 0.24) mm and (10.34 +/- 3.12) mm respectively. External diameter and length of right V , VII, VIII, IX cranial nerves are (2.52 +/- 0.86) mm and (6.91 +/- 2.66) mm, (1.14 +/- 0.31) mm and (10 +/- 2.96) mm, (2.13 +/- 0.63) m and (10.09 +/- 2.93) mm, (0.790.29) mm and (10.17 +/- 3.06) mm. intermedius nerve locate between facial nerve and acoustic nerve, external diameter of intermedius nerve is (0.47 +/- 0.91) mm (left) and (0.37 +/- 0.07) mm (right). Length of vagal nerve is (10.44 +/- 2.57) mm (left), (9.91 +/- 2.91) mm (right), rootlets of f vagal nerve is 6.37 +/- 2.26 (left) and 6.33 +/- 2.38 (right). external diameter of accessory nerve is (0.76 +/- 0.16) mm (left) and (0.81 +/- 0.19) mm (right). CONCLUSION This study provide anatomical data for retrosigmoid approach in the cerebellopontine angle region.
Adult ; Cerebellopontine Angle ; anatomy & histology ; Cranial Nerves ; anatomy & histology ; Cranial Sinuses ; anatomy & histology ; surgery ; Facial Nerve ; anatomy & histology ; Glossopharyngeal Nerve ; anatomy & histology ; Humans ; Vestibulocochlear Nerve ; anatomy & histology

OBJECTIVE: To observe the effects of recipes for replenishing qi and activating blood on p16, p21, proliferating cell nuclear antigen (PCNA), cyclin D1 and cyclin E gene expressions in the liver of aging rats. METHODS: A recipe for replenishing qi and a recipe for activating blood were administered to aging rats respectively, and the effects of the above recipes on the expressions of senescence related genes (p16, p21, PCNA, cyclin D1 and cyclin E) were examined by RT-PCR and Western blotting methods. RESULTS: The expressions of p16, p21 and cyclin D1 mRNAs and proteins in the liver of the untreated aging rats were up-regulated, while the expressions of PCNA and cyclin E mRNAs and proteins decreased. As compared with the untreated aging rats, both recipes could down-regulate the expressions of cyclin D1 mRNA and protein and up-regulate the expressions of cyclin E mRNA and protein, but had no obvious effects on the expressions of mRNAs and proteins of p16, p21 and PCNA. CONCLUSION: Recipes for replenishing qi and activating blood can improve the liver cell proliferation of aging rats via down-regulating the expressions of cyclin D1 mRNA and protein and up-regulating the expressions of cyclin E mRNA and protein.

BACKGROUNDTo evaluate and compare the effects and toxicity of the domestic product of recombinant mutant human tumor necrosis factor (rmhTNF) combined with chemotherapy and chemotherapy alone in the treatment of patients with non-small cell lung cancer (NSCLC).

METHODSTwo hundred patients with NSCLC in multicenter were randomly devided into trial group (150 cases) and control group (50 cases). Chemotherapy with CAP regimen was given to the patients. Meanwhile, rmhTNF injection of 4×10⁶U/m² was also given from the 1st to 7th days, the 11th to 17th days on the chemotherapy cycle in the trial group. The control patients received chemotherapy alone. Twenty-one days were as a cycle, 2 cycles were given to each patient. The chemotherapeutic effects and toxicity were observed and compared between the two groups after the therapy.

RESULTSof the 200 patients, 5 cases in the trial group and 3 cases in the control group were out of the trial because of economy. The other 192 cases (145 cases in the trial group and 47 cases in the control group) could be analyzed and evaluated the clinical effects and toxicity. The response rate of chemotherapy was 46.90% (68/145) in the trial group and 17.02% (8/47) in the control group respectively ( P =0.001). The KPS scores was 86.02±9.74 in the trial group, and 80.14±9.10 in the control group ( P =0.025). No significant difference of degree III+IV toxicity was observed between the two groups ( P > 0.05). The side effects related to rmhTNF included slight fever, cold-like symptoms, pain and red and swelling in the injection site. All of them were mild and didn't need any treatment and disappeared after the therapy. There were no severe abnormality of liver and kidney function and ECG in both groups.

CONCLUSIONSThe results demonstrate that the effects of domestic rmhTNF combined with chemotherapy are remarkably higher than that of chemotherapy alone in the treatment of NSCLC. rmhTNF can increase the sensitivity to chemotherapy and improve the quality of life of the patients with slight toxicity. Hence rmhTNF is worth expanding clinical use.