Objective To study the effects of rhubarb on the mitogen-activated protein kinase (MAPK) sig-naling transducfon pathway in rats with severe acute pancreatitis (SAP),and to investigate the treatment mecha-nism of rhubarb on SAP. Method One hundred SD rats were provided by from the Animal Center of Nanjing Uni-versity. All animals were randomly divided into sham operation (n=33), SAP (n=33) and rhubarb groups (n=34). SAP model was induced by retrograde injection of 5% sodittm taurocholate. Rhubarb was given with 10% rhubarb decoction (2 mi/100 g) at the time of pancreafitis induction in the rhubarb groups. At 1, 3, 6, and 12 h after the models were established,animals were killed. MAPK activity in pancreatic tissue was examined by West-em blotting and the mRNA levels of TNF-α and IL-6 in pancreatic tissues were detected by RT-PCR. All data were analyzed by SPSS statistical software and statistical differences between values from two sroups were determined by the Student's t -test. Results MAPK activity, TNF-α and IL-6 mRNA levels in pancreatic tissues were signifi-cantly enhanced in the SAP group compared with the sham operation group (all P<0.01). Rhubarb treatment markedlyinhibited MAPK activation,TNF-α,IL 6 mRNA (all p<0.01). Conclusions Rhubarb can alleviate the inflammatory response of SAP by down-regulating MAPK activity.

ObjectiveTo explore the viability,security and the curative effect of ultrasonic guided percutaneous portal vein puncture on autologous stem cells transplantation for patients with decompensated liver cirrhosis.MethodsSixty-four patients of decompensated liver cirrhosis with ascites were treated by autologous stem cell transplantation through ultrasonic guided percutaneous portal vein puncture.The clinical symptoms after transplantation,postoperative adverse reaction,the results of liver function tests were observed.Results After stem cells transplantation,patients' liver function got better obviously,laboratory indicators were better,postoperative adverse reaction and complication were less than those of 87patients treated with autologous stem cell transplantation into hepatic artery multilevel branch by pipe(P＜0.05).Conclusions Ultrasound guided percutaneous portal vein puncture on autologous stem cells transplantation is the viability,security,less side effect and effective for treatment patients with decompensated liver cirrhosis.It can be used as the clinical treatment methods for patients with middle-late liver cirrhosis.

Objective:To analyze the genomic characteristics and variations of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) derived from an outbreak in inbound flight in Aug, 2022 in Beijing, and provide reference data for disease prevention and control and risk assessment.Methods:Fifty respiratory tract specimens from all cases in this outbreak were collected. The digital PCR (ddPCR) was used to determine the viral loads of the specimens. The full genome of the viruses were sequenced by Next-generation sequencing. Then analuses were performed on the genetic identity, variations and phylogenesis.Results:The median of viral loads in all 50 samples were 5.57×10 4 copies/ml and 5.85×10 4 copies/ml, for ORF1ab gene and N gene, respectively. A total of 46 SARS-CoV-2 genomes were obtained, which all belonged to Omicron/BA.5. Two genome clusters were observed, involving 21 and 7 cases, with a nucleotide sequence identities of 99.993% and 99.997%, respectively. Conclusions:The studied outbreak was composed of two main clusters and other individual cases with Omicron/BA.5 virus overseas.

Objective:To investigate the phylogenetic and antigenic characteristics of hemagglutinin (HA) gene of influenza B/Victoria lineage (BV) viruses in Beijing during the 2021-2022 influenza surveillance season, and to analyze whether the circulating BV viruses match the vaccine strain.Methods:Pharyngeal swab specimens from influenza like-illness (ILI) cases in the 2021-2022 influenza surveillance season were collected from surveillance network labs in Beijing and cultured in MDCK cells and chicken embryo to isolate BV viruses. Nucleic acids of the viruses were extracted, and the HA gene was amplified and sequenced. The nucleotide and amino acid sequence identity of the HA gene was analyzed using MEGA5.0 software. A phylogenetic tree of HA gene was constructed using the maximum likelihood method. The N-glycosylation sites in HA were predicted online. Three-dimensional structure of HA was constructed using SWISS-MODEL homologous modeling. Hemagglutination inhibition (HI) test was performed to analyze the antigenicity of BV viruses.Results:A total of 402 BV viruses were collected and 58 strains with full-length HA gene sequences were chosen for further analysis. Compared with the HA gene of this year′s vaccine strain (B/Washington/02/2019), there were 27 amino acid mutations, 11 of which were located in four different antigenic determinants. The phylogenetic analysis revealed that three subgroups of 1A.3, 1A.3a1, and 1A.3a2 co-circulated in Beijing with 54 strains (54/58, 93.10%) clustered to the Clade 1A.3a2, two strains (2/58, 3.45%) clustered to the Clade 1A.3a1, and two strains (2/58, 3.45%) in the same subgroup (Clade 1A.3) as the vaccine component BV strain in 2021-2022. Compared with the vaccine strain (B/Washington/02/2019), two BV strains had an additional N-glycosylation site at residue 197, while the other 56 strains showed no change in N-glycosylation sites. Antigenic analysis showed that 35 BV strains (35/58, 60.34%) were antigenically similar to the vaccine strain and 23 strains (23/58, 39.66%) were low-response strains.Conclusions:Three subgroups of BV viruses co-circulated in Beijing during the 2021-2022 influenza surveillance season. The predominant subgroup was Clade 1A.3a2 (93.10%), showing a certain genetic distance with the vaccine strain (B/Washington/02/2019). Nearly 40% (39.66%) of the viruses were low-response strains. This study indicated that continuous monitoring of the variations of influenza epidemic strains and timely providing laboratory basis for screening vaccine component strains were the basic technical guarantee for coping with influenza pandemic.

Objective:To disclose phylogenetic and antigenic characteristics of hemagglutinin (HA) gene of influenza B virus (Victoria) (BV) in the 2023-2024 influenza surveillance season in Beijing, and understand the matching with influenza vaccine component strain.Methods:Pharyngeal swab specimens from influenza like-illness (ILI) in the 2023-2024 influenza surveillance season were collected from surveillance network labs in Beijing and BV strains were isolated through MDCK or chicken embryo culture. After extracting nucleic acid, HA gene was amplified and sequenced. The nucleotide and amino acid sequence identity were conducted and the maximum likelihood method in Mega 5.0 software was used to construct the phylogenetic tree of HA gene. N-glycosylation sites of HA were performed online. Furthermore, three-dimensional structure of HA was available from SWISS-MODEL homologous modeling. Hemagglutination inhibition (HI) tests were performed to analyze antigenic characteristics of HA of BV strains.Results:Fifty-four BV strains were randomly selected to be analyzed further. Compared with the HA gene of this influenza season vaccine strain (B/Austria/1359417/2021), there are three amino acid mutations among all BV strains, two of which are located in two different antigenic determinants. Furthermore, the phylogenetic tree analysis revealed that only one subgroup of 1A.3a.2 was circulating simultaneously. All BV strains are located in Clade 1A.3a.2 subgroup, and in the same subgroup with that of the vaccine component BV strain in 2023-2024. All BV strains have the same glycosylation sites as that of the vaccine component BV strain in 2023-2024. Antigenic analysis showed that all BV strains were antigenically similar with its vaccine strain.Conclusions:In the 2023-2024 influenza surveillance season, the prevalent BV strains in the population in Beijing city are located in Clade 1A. 3a. 2 subgroup. The antigen matching between BV epidemic strains and vaccine BV components is relatively high during this surveillance season.

Objective This study evaluated two isothermal amplification methods for their ability to identify targeted virus in the clinical samples,which provided a reference for other researchers using NGS in clinical diagnostics.Methods Nasopharyngeal swabs from two patients with severe pneumonia were amplified with Single Primer Isothermal Amplification (SPIA) method and Multiple Displacement Amplification (MDA) approaches and further proceeded to NGS and metagenomics analysis to identify targeted virus.Results The SPIA produced shorter fragments than MDA,and had a lower amplification efficiency.The SPIA successfully identified targeted viruses in the two clinical samples tested,while MDA failed to detect that in one of the samples.Conclusions Both SPIA and MDA could be applied to pathogen identification using NGS in the clinical diagnostics.SPIA possessed lower amplification efficiency and higher economic cost,but had higher capacity to discover the causing agents in the clinical sample.

Objective:To establish a nested PCR method to detect the 2019 novel coronavirus (2019-nCoV), as a supplement to the real-time fluorescent PCR method, and discuss the preliminary application value of this method in clinical diagnosis.Methods:According to the conservative sequences of the 2019-nCoV gene, the nested PCR primers including N gene and S gene, were designed on line. By optimizing the nested PCR reaction systems, the qualitative detection was established by testing N gene and sequencing its PCR product while the preliminary type identification was established by testing S gene and sequencing its PCR product. The sensitivity was evaluated by the gradient dilution of 2019-nCoV positive samples’ nucleic acid and the specificity was evaluated by detecting the human coronavirus OC43, 229E, HKU1, NL63, influenza virus positive samples. The established method was applied to 15 samples with Ct >33 and 15 samples with Ct <33 screened by real-time fluorescent PCR, and the positive amplification result were sequenced and analyzed to verify the result. Results:The established nested PCR method could amplify specific bands of 355 bp N gene fragment and 449 bp S gene fragment. No amplifications occurred in other human coronaviruses samples including 229E、OC43、HKU1、NL63 or in influenza virus samples including H3N2, H1N1(pdm) and B. The minimum detection limit of the N gene fragment could reach Ct value about 37.21. Among the 30 COVID-19 positive samples, the N gene positive coincidence rate detected by nested PCR was 100% (30/30); the S gene positive coincidence rate reached 60% (18/30). 28 samples’ sequences of N gene fragment were completely consistent with 2019-nCoV by BLAST, and the characteristic result of site mutations of 12 samples’ S gene was obtained. Conclusions:A nested PCR method for the specific detection of 2019-nCoV was established, and some characteristic mutations on S gene could be analyzed by sequencing the PCR amplified products. It could be used as a supplement to the real-time fluorescent PCR method.

Objective:To investigate frequency and position characteristics of the vestibular dysfunction in vestublar neuritis patients. Method:Colaric test (CT), head impulse test (HIT), cervical vestibular evoked myogenic potential (cVEMP) and ocular vestibular evoked myogenic potential (oVEMP) were applied in 43 vestublar neuritis patients to assess their vestublar dysfunction. Superior vestublar nerve (S-VN), inferior vestibular nerve (I-VN), total vestibular nerve (T-VN) and each vestibular end organ incidence rate were calculated and statistically analyzed. Result:CT incidence rate (93.0%) was statistically higher than that of HIT (72.1%) (P<0.01). Total frequency incidence rate (72.1%) was statistically higher than that of low frequency (20.9%) (P<0.01). No high frequency only case was observed. The incidence rate of S-VN only, I-VN only and T-VN was 44.2%, 4.7% and 51.2% respectively. Among them, the incidence rate of I-VN was significantly lower than the others (P<0.01). The incidence rate of vestibular end organs was 17.4% (S-SCC), 44.2% (H-SCC), 20.9% (P-SCC), 39.5% (utricule) and 26.7% (saccule) respectively. The incidence rate of H-SCC was remarkably higher than the other semicircular canals (P<0.01). The difference between utricule and saccule was not statistically significant. Conclusion:The semicricular canal dysfunction in vestibular neuritis patients mainly involves total frequency of vestibular function, low frequency is more common than high frequency. Total vestibular nerve and single S-VN are mostly involved in vestibular neuritis.