Objective To investigate the modeling elements of various types of animal models for acute liver injury,and to provide references and suggestions to establish and evaluate animal models of acute liver injury(ALI).Methods The animal experimental literature of ALI from 2002 to 2022 was searched in the databases of the China Knowledge Network,WanFang,Chongqing Vip(VIP),Chinese Medical Journal Full Text Data(Yiigle),and PubMed.The animal species,positive control drugs,modeling method,modeling drugs,and drug administration of the animal models of ALI in the literature were summarized.The result were analyzed using Excel,SPSS Modeler 18.0,and Cytoscape 3.8.2.Results A total of 896 articles were included in the databases.The most used animal models for ALI were male KM mice.The modeling method were mainly chemical liver injury,alcoholic liver injury,drug-related liver injury,and immune liver injury.①The corresponding main modeling method were intraperitoneal injection of 10 mL/kg of 0.1％CC14 in vegetable oil at 24 h before experiments,②gavage of 12.0 mL/kg of 50.0％～56.0％ethanol at 16 h before experiments,③intraperitoneal injection of 300 mg/kg APAP at 24 h before experiments,④tail vein injection of 20 mg/kg Con A at 8 h before experiments.Evaluation of the models was based on liver pathological indexes as the gold standard combined with biochemical indexes of serum ALT,AST,and SOD and MDA contents and activities in liver tissue homogenate as direct indicators.Conclusions Because the causes of ALI vary in clinical practice,the preparation of animal models of ALI should be based on the specific study content and characteristics,and the corresponding modeling method should be selected.

ObjectiveTo compare the effects of topical application of Modified Sanhuang Powder （加味三黄散， MSP） combined with cold compression versus cold compression alone on swelling and pain after knee arthroscopy through a retrospective cohort study. MethodsMedical records of 134 patients with knee arthroscopy-induced knee swelling and pain were divided into non-exposure group （51 cases） and exposure group （83 cases） based on whether they used MSP for external application after surgery. The non-exposure group received simple cold compression therapy in addition to functional exercise and routine treatment after surgery， while the exposure group received topical MSP on the basis of what were given in the non-exposure group. The Visual Analog Scale （VAS） scores were compared between the two groups before and 7 days after treatment， and knee swelling measurements were taken before and 3， 5， and 7 days after treatment. The clinical effective rate was compared between the two groups. ResultsThe VAS scores in both groups were lower after treatment （P＜0.05）， and the exposure group had lower scores than the non-exposure group （P＜0.05）. On the 3rd， 5th， and 7th days of treatment， the scores on swelling at 2 cm above the superior pole of the patella， at the midline of the patella， and 5 cm below the inferior pole of the patella significantly decreased after treatment in both groups （P＜0.05）， and the exposure group had lower scores than the non-exposure group （P＜0.05）. The total clinical effective rate in the exposure group was 91.56% （76/83）， which was higher than 78.43% （40/51） in the non-exposure group （P＜0.05）. ConclusionTopical application of MSP combined with cold compression is effective in relieving postoperative swelling and pain after knee arthroscopy and is superior to cold compress alone.

Objective:To study the genomic sequence of Coxsackievirus A8 (CV-A8) associated with hand, foot and mouth disease (HFMD) from 2013 to 2018 in China and to analyze the genetic evolution of each coding region of the full-length genome.Methods:The genome sequences of 11 CV-A8 strains isolated from patients with HFMD in different regions of China from 2013 to 2018 were determined. Sequence alignment and genetic evolution analysis were performed by Sequencher 5.0 and MEGA 7.0 software, etc.Results:Sequence alignment showed that the genome length of 11 CV-A8 strains ranged from 7 393 bp to 7 400 bp. There was no base insertion or deletion in the coding region compared with the prototype strain, but there were individual base insertion or deletion in the non-coding region. The nucleotide and amino acid similarities in the VP1 region of 11 CV-A8 strains were 78.3%-98.6% and 92.6%-99.7%, respectively, and the nucleotide and amino acid sequences identities with the CV-A8 prototype strain were 78.3%-98.2% and 92.6%-99.7%, respectively. Based on the phylogenetic analysis of VP1 region sequences, the CV-A8 can be divided into five genotypes: A, B, C, D and E. The 11 CV-A8 strains in this study belonged to genotypes C (1 strain), D (2 strains) and E (8 strains). The nucleotide and amino acid similarities of 11 CV-A8 full-length genomes were 81.3%-98.8% and 95.9%-99.5%, respectively. The phylogenetic tree of the P2 region showed that the eight E genotypes CV-A8 had the closest evolutionary distance with CV-A4, CV-A14, and CV-A16. The phylogenetic tree of the P3 region showed that the eight E genotypes CV-A8 had a close evolutionary distance with CV-A5, CV-A16, CV-A14 and CV-A4.Conclusions:The 11 CV-A8 stains in this study showed significant intra-genotype diversity in capsid region and recombinant diversity in non-capsid region which indicated that CV-A8 quasispecies were still undergoing dynamics variation. CV-A8 may become an important pathogen of HFMD and the monitoring of CV-A8 needs to be further strengthened.

Objective:To evaluate the effects of vaccine switch on the levels of neutralizing antibody (NA) of poliovirus in children aged<12 years.Methods:Subjects aged<12 years from 9 districts in Fujian province were chosen by stratified cluster and complete random sampling method. Blood samples were collected for testing NA of poliovirus by microcell neutralization.Results:A total of 2 134 subjects aged<12 years were selected. The positive rate of NA against PVⅠand Ⅲ were 98.64% and 95.83%; and geometric mean titers (GMTs) were 1∶259.35 and 1∶105.14, respectively. The GMTs presented a trend of decreasing as age increased. Compared to trivalent oral poliovirus vaccine (tOPV), bivalent oral poliovirus vaccine (bOPV) and inactivated poliovirus vaccine (IPV) induced higher GMTs of NA against poliovirusⅠand Ⅲ respectively. Among 182 children aged<5 years, the positive rate of NA against PVⅠ, Ⅱ, Ⅲ were 97.25%, 76.37% and 92.86%. There were statistical differences among the three types ( χ2=44.44, P=0.000). The rate of NA against PVⅡwas significantly lower than those ofⅠand Ⅲ (Ⅱ vs. I: χ2=34.65, P=0.000; Ⅱ vs. Ⅲ: χ2=18.99, P=0.000). And the GMTs of NA against PV Ⅰ, Ⅱ, Ⅲ were 1∶368.96、1∶23.06 and 1∶183.10, which were significantly different ( F=156.54, P=0.000). The GMT of PVⅠwas the highest, PV Ⅲ was the second, PVⅡ was lowest (pairwise comparison showed values of P all were 0.000). The analysis of general linear model showed that the interval between the date of last immunization and the date of sample collection would affect the GMTs of PVⅠand Ⅲ, different vaccine models would affect only the GMT of PV Ⅰ. The age maybe was a confounding factor. But no factors would affect the GMT of PV Ⅱ. Conclusions:After the vaccine switch, the level of NA against PVⅠand PV Ⅲ were still maintaining high level, but the level of PVⅡamong children<5 years was at relatively low level, which indicated that the surveillance should be strengthened.

COVID-19 has swept globally and Pakistan is no exception.To investigate the initial introductions and transmissions of the SARS-CoV-2 in Pakistan,we performed the largest genomic epidemiology study of COVID-19 in Pakistan and generated 150 complete SARS-CoV-2 genome sequences from samples collected from March 16 to June 1,2020.We identified a total of 347 mutated positions,31 of which were over-represented in Pakistan.Meanwhile,we found over 1000 intra-host single-nucleotide variants(iSNVs).Several of them occurred concurrently,indicating possible interactions among them or coevolution.Some of the high-frequency iSNVs in Pakistan were not observed in the global population,suggesting strong purifying selections.The genomic epidemiology revealed five distinctive spreading clusters.The largest cluster consisted of 74 viruses which were derived from different geographic locations of Pakistan and formed a deep hierarchical structure,indicating an extensive and persistent nation-wide transmission of the virus that was probably attributed to a signature mutation(G8371T in ORF 1ab)of this cluster.Further-more,28 putative international introductions were identified,several of which are consistent with the epidemiological investigations.In all,this study has inferred the possible pathways of introduc-tions and transmissions of SARS-CoV-2 in Pakistan,which could aid ongoing and future viral surveillance and COVID-19 control.

Objective To evaluate the inactivation effect of heat inactivation and three common chemical disinfectants on enterovirus group A 71 (EV-A71),provide basic data for disinfection work after laboratory activities in enterovirus secondary biosafety laboratory.Methods EV-A71 virus was treated at different temperatures and different disinfectant concentrations according to different time.The inactivated virus was inoculated into 96-well cell culture plates,and the cytopathic effect was observed and recorded.Finally,the virus titer of the virus treatment solution was calculated.Results After inactivation at 56 ℃ for 30 min,the inactivation logarithm of EV-A71 was more than 6,and inactivation at 65 ℃ for 30 rin,70 ℃ for 10 min,no infection was observed.Three chemical disinfectants could effectively inactivate a certain titer of EV-A71 at a certain concentration and treatment time.Conclusions The inactivation effect of EV-A71 virus is related to factors such as thermal inactivation temperature,chemical disinfectant concentration,and treatment time.Studying the inactivation curve of EV-A71 is beneficial to the selection of inactivation conditions in different inactivation environments.

AIM: To investigate the apoptosis and molecular mechanism of human hepatocellular carcinoma HepG2 cells induced by ginsenoside Rh4.METHODS: Human hepatocellular carcinoma HepG2 cells were treated with ginsenoside Rh4 at doses of 10, 20 and 40 μmol/L, and the inhibitory effect of ginsenoside Rh4 on HepG2 cell viability was measured by MTT assay.The apoptotic rate of HepG2 cells was analyzed by flow cytometry.The morphological changes of the HepG2 cells were observed by Hoechst 33258 and TUNEL staining.The expression of apoptosis-related proteins Bax, Bcl-2, caspase-3 and caspase-9 was determined by Western blot.RESULTS: Ginsenoside Rh4 promoted apoptosis of HepG2 cells in a dose-dependent manner.TUNEL and Hoechst 33258 staining showed that the cells appeared obvious shrinking, swelling and rupture after treated with ginsenoside Rh4 for 24 h.The results of Western blot showed that with the increasing concentrations of ginsenoside Rh4, the expression of pro-apoptotic proteins Bax, cleaved caspase-3 and caspase-9 increased, while anti-apoptotic protein Bcl-2 decreased gradually.CONCLUSION: Ginsenoside Rh4 induces apoptosis of human hepatocellular carcinoma HepG2 cells, and the main mechanism may be related to down-regulation of Bcl-2 and up-regulation of Bax, cleaved caspase-3, and caspase-9.

Objective To study the relationship between different syndromes of AIDS dementia complex (ADC) and different disease severity, age, CD4+ T cell count and infection. Methods Totally 186 patients with ADC were classified into different syndrome types, and the distribution in different degree of disease, different age, different CD4+T cell count and different routes of infection was analyzed. Results There were 48, 51, 15, 37 and 35 cases of deficiency of kidney and marrow, yin deficiency of liver and kidney, deficiency of heart and spleen, syndrome of phlegm obstruction, syndrome of qi deficiency and blood stasis, respectively. Moderate and severe degrees with yin deficiency of liver and kidney were more common. There was statistical significance in the distribution of different syndromes in different degree of disease (χ2=82.495, P=0.000). Deficiency of kidney and marrow, yin deficiency of liver and kidney were more common in different age groups. The distribution of the syndrome types in different age groups was statistically significant (χ2=72.710, P=0.000), the patients were mainly in two age groups of>50–60 years old and>60 years old. The distribution of the syndrom types in diffenrent CD4+T cell count stratum was statistically significant (χ2=66.778, P=0.000). Blood pathway infection mainly included deficiency of kidney and marrow and syndrome of qi deficiency and blood stasis, sexual pathogens mainly yin deficiency of liver and kidney. Conclusion CD4+T cells layers, age group, progression of disease and transmission way are the influencing factors of syndrom type.

Objective To explore the effect of infrared spectral analysis in analyzing of the chemical composition of renal staghorn calculi and its relationship with urinary tract infections. Methods From June 2014 to June 2016, the clinical data of 186 patients with renal staghorn calculi were collected. The stone composition were analyzed by infrared spectroscopy and traditional chemical titration, and the stones infection were detected by microbial analysis system. The relation between stones infection, urinary tract infection and stone composition were analyzed. Results The results of infrared spectroscopy and traditional chemical titration in detecting renal staghorn calculi ingredient had no significant differences (P>0.05). In 186 patients, 56 patients (30.11%) was in infected group, and 130 patients (69.89%)was in non-infected group. The abnormal urine rate, urinary tract infection rate, medistream urine positive infection rate and cotton swabs positive infection rate in infected group were was significantly higher than those in non-infected group: 73.21%(41/56) vs. 50.77%(66/130), 19.64%(11/56) vs. 3.85%(5/130), 50.00%(28/56) vs. 6.15%(8/130), 67.86%(38/56) vs. 8.46%(11/130), P<0.01. The carbonate apatite stones rate and six water magnesium ammonium phosphate rate in infected group were significantly higher than those in non-infected group: 21.43%(12/56) vs. 5.37%(7/130), 57.14%(32/56) vs. 2.31%(3/130), P<0.01. The calcium oxalate rate and uric acid rate in non-infected group were significantly higher than those in infected group:50.00%(65/130) vs. 5.36%(3/56), 24.62%(32/130) vs. 1.79%(1/56), P<0.01. Conclusions Analysis of staghorn calculi ingredient caused by urinary bacterial infection with infrared spectroscopy is simple, reliable and easy to operate. It is important for postoperative infection prevention.

Objecve To observe the clinical effect and safety of the Chinese medicine syndrome differentiation combined with HAART for the ADC(acquried immune deficiency syndrom dementia complex). Methods A total of 80 patients with ADC were divided into the treatment group and control group based on random number table, 40 in each group. The patients in the control group were treated by highly active anti-retrovital therapy (HAART). The patients in the treatment group were treated with TCM treatment on the based of the control group. Both groups received the treatment for 3 months.These outcomes were measured: TCM syndrome integral, mini mental state examination(MMSE), daily behavior scale(ADL), change of clinical stage, and adverse reactions. Results The effect rate of treatment group was 82.5%, which was significant higher than 65% of the control group (χ2=8.115,P=0.024). After the treatment, the ADL integral of the treatment group (37.69 ± 5.31vs.33.67 ± 5.16;t=2.528,P=0.021) was significantly higher than that before the treatment; and the ADL integral of the control group(36.96 ± 5.52vs.34.54 ± 4.98;t=2.747,P=0.027) was significantly higher than that before the treatment.But there was no significant difference between the two groups after the treatment (t=2.003,P=0.139). After the treatment, the MMSE integral of the treatment group (24.76 ± 4.43 vs.19.97 ± 5.46;t=1.006,P=0.013) was significantly higher than that before the treatment; the MMSE integral of the control group(24.65 ± 4.36 vs. 20.11 ± 4.87;t=1.035,P=0.014) was significantly higher than that before the treatment. But there was no significant difference between the two groups after the treatment (t=0.953, P=0.347).There was no significant difference between the two groups in the clinical stage change (phase1χ2=1.231,P=0.954; phase2χ2=2.726,P=1.053). There was no adverse reaction in the two groups during the treatment.Conclusions The Traditional Chinese medcine combined with HAART was better than HAART alonein the treatment of ADC.