Objective:To investigate the role of complement C3a receptor in the diabetic nephropathy pathogenesis of db/db mice, and to provide a new target for prevention and treatment of diabetic nephropathy.Methods:Twelve 8-week-old male mice with type 2 diabetes mellitus (db/db mice) and 6 wild-type (db/m) mice were reared in the special pathogen free environment. The mice were grouped into db/m group, db/db group and C3a receptor antagonist group, with 6 mice in each group. db/db model mice were intraperitoneally injected with C3a receptor antagonist (SB290157, 10 mg/kg) once every two days for 8 weeks in C3a receptor antagonist group. Blood and urine samples were collected, and body weight of mice, fasting blood glucose, serum creatinine, blood urea nitrogen, urinary microalbumin/urinary creatinine and urinary N-acetyl-β- D-glucosaminidase (NAG) were detected. Renal tissues were collected, and HE, PAS and Masson stainings were used to observe the pathological changes. Immunohistochemistry, immunofluorescence and Western blotting were used to detect the protein expression levels of C3 and C3a receptor. Western blotting was used to analyze the protein expression levels of kidney injury molecule-1 (Kim-1), α-smooth muscle actin (α-SMA), zonula occluden-1 (ZO-1), vimentin and E-cadherin in renal tissues. Immunofluorescence was used to analyze the protein expression levels and distribution of α-SMA, ZO-1 and Kim-1, and immunohistochemistry was used to analyze the protein expression levels of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α). TUNEL assay was used to detect apoptotic cells in renal tissues. Results:Compared with db/m group, body weight, fasting blood glucose, urinary microalbumin/urinary creatinine and urinary NAG in db/db group were significantly higher, while these indicators in C3a receptor antagonist group were slightly lower than those in db/db group (all P<0.01). There were no significant differences in serum creatinine and blood urea nitrogen among the three groups (all P>0.01). Compared with db/m group, db/db group had glomerular hypertrophy, necrosis and exfoliation of renal tubular epithelial cells, and dilation of renal tubules, and C3 and C3a receptor protein expression levels were higher (both P<0.01). Compared with db/db group, C3a receptor antagonist group had less glomerular lesions, mild necrosis of renal tubular epithelial cells and less tubular dilation. Compared with db/m group, the protein expression levels of Kim-1, IL-1 and TNF-α in kidney tissues of db/db group were significantly higher, while Kim-1, IL-1 and TNF-α in C3a receptor antagonist group were significantly lower than those in db/db group (all P<0.01). Compared with db/m group, the protein expression levels of α-SMA and vimentin of renal tubular epithelial cells in db/db group were significantly higher, while the protein expression levels of ZO-1 and E-cadherin were significantly lower (all P<0.01). Compared with db/db group, the protein expression levels of α-SMA and vimentin of renal tubular epithelial cells in C3a receptor antagonist group were significantly lower, and the protein expression levels of ZO-1 and E-cadherin were significantly higher (all P<0.01). Compared with db/m group, the number of apoptotic cells of kidney tissues in db/db group was increased, while the number of apoptotic cells in C3a receptor antagonist group was reduced compared with db/db group. Conclusions:The expression levels of C3 and C3a receptor of kidney tissues in db/db mice are significantly increased. Antagonistic C3a receptor can reduce the body weight, blood glucose, urinary microalbumin/urinary creatinine and urinary NAG, alleviate renal pathological injury, inhibit renal tissue inflammation, apoptosis and renal tubule epithelial-mesenchymal transition in db/db mice.

Objective:To confirm the cross-provincial long-distance transmission events of Coronavirus Disease 2019 (COVID-19) by lorry drivers, the origin of infections of the cases and the transmission routes were tracked.Methods:Nasopharyngeal swab specimens from five lorry driver cases of COVID-19, found in Zhangzhou city in March, 2022 when the local outbreaks occurred in adjacent Quanzhou city, Fujian province, were collected to perform 2019 novel coronavirus (2019-nCoV) targeted genome amplification and followed by next-generation sequencing. The sequences were submitted to online 2019-nCoV analysis platforms to classify the type of variant and mutation sites. Phylogenetic tree for the viruses were constructed by phylogenetic analysis software. Combined with the epidemiological investigation, the origin of infections of the cases and the transmission routes were deduced.Results:Five complete genome sequences, with 29 770-29 839 bp in length and 99.53% average genomic coverage, of 2019-nCoV were successfully obtained. The viruses were all Omicron variants and further divided into three different subclades of BA.2. Of the five strains of 2019-nCoV, three were highly similar to the viruses of two distinct lineages co-circulated in Quanzhou city during the period of local outbreak of COVID-19, respectively. Phylogenetic analysis also revealed that the viruses from three infected lorry drivers were highly homologous to that from local outbreaks in Quanzhou city. The viruses from the rest two cases had seven to fourteen nucleotide mutations (corresponding to 5-7 amino acid substitutions) when compared with the viruses in local outbreaks in Quanzhou city, which excluded the involvement of the two cases into the transmission chains of local outbreaks. Combined with the field epidemiological investigations, the result revealed that the origin of infection of 2019-nCoV of the two sporadic lorry driver cases was outside of Fujian province.Conclusions:With the aid of high-throughput sequencing and bioinformatics technology combined with field epidemiological investigations, we speculated in this study that at least three origins of infection of 2019-nCoV in five lorry driver cases and cross-provincial long-distance transmission via two sporadic cases infected outside Fujian province when they returned.

Objective:To investigate the whole genome characteristics and mutation of imported 2019-nCoV and trace potential source at the genomics level, the viral genomes from the specimens of a cluster of COVID-19 cases which were imported from a freighter were directly sequenced and determined by high-throughput sequencing technology and bioinformatics analysis.Methods:Throat swab specimens from the 8 confirmed cases of COVID-19 from the same freighter were collected to perform the whole genome sequencing for 2019-nCoV using the targeted genome amplification, combined with Ion S5 next-generation sequencing (NGS) technology. Varieties of online virus analysis platforms was used to classify the viruses and analyze mutation sites in the whole genome. Phylogenetic analysis software was used to construct a phylogenetic tree for the viruses, combined with the epidemiological data of the case, to speculate about the source of the viruses.Results:Eight complete genome sequences of 2019-nCoV was successfully obtained. The complete genomes of the viruses were 29, 822-29, 865 bp in length, with the average sequencing depth of 11 928×-33 588× and the coverage of 99.73%-99.87%; the result of Pangolin classification showed that all the eight 2019-nCoV genomes belong to VOC/Delta (B.1.617.2) lineage. The result of whole genome mutation analysis showed that compared with the Wuhan reference strain, the median number of nucleotide mutations in the eight 2019-nCoV genome sequences was 35 (31 to 38), and the median number of amino acid mutations was 26 (24-28); the mutation sites were distributed in 8 gene coding regions (ORF1a, ORF1b, S, ORF3a, M, ORF7a, ORF8, N). Further analysis revealed that eight 2019-nCoV genomes contained 23 characteristic mutation sites belonging to the Delta (B.1.617.2·AY.2) variants of 2019-nCoV. Since the mutation sites among the eight 2019-nCoV genomes were not completely overlapped, and the epidemiological survey report showed that the freighter stopped at multiple ports and had personnel alternation, it was speculated that the clustered COVID-19 cases might have different origins. Phylogenetic analysis showed that all the eight 2019-nCoV genomes were on the AY.2 sub-lineage of the B. 1.617.2 lineage. This result was consistent with the result of Pangolin classification and mutation analysis.Conclusions:In this study, 8 whole genome sequences of Delta variants were obtained through NGS technology from the clustered COVID-19 cases which were imported from a freighter. The sequencing method and analysis result in this study could provide reference for the 2019-nCoV mutation analysis and tracing the source of the COVID-19 cases for the prevention and control of the COVID-19 epidemic.

Through the description of the content and design of "first-class course of virtual simulation of flow cytometry application", combined with the application in experimental teaching of different subjects, this paper expounds the teaching effect and students' evaluation of the blended learning method of "virtual simulation experiment of flow cytometry application" combined with offline experimental content by means of questionnaire survey, and puts forward some views and opinions on the construction of the virtual simulation experiment teaching content system.

Objective To evaluate the clinical efficacy and safety of vancomycin and linezolid for the treatment of gram-positive neonatal bacterial sepsis.Method The data of neonates diagnosed as grampositive bacterial sepsis in neonatology department of Suzhou Municipal Hospital from June 2009 to December 2015 were retrospectively collected.These neonates were divided into vancomycin group and linezolid group.Propensity score matching (PSM) on baseline variables was used to balance the two groups by identifying a comparable group of neonates who received vancomycin and linezolid therapy.Clinical and microbiologic success rates were compared by chi-square test,and changes of laboratory parameters before and after treatment at the end of treatment were then directly compared by rank-sum test between the matched groups.In vancomycin group,correlation between trough concentration of vancomycin and clinical efficacy were evaluated.Result Totally 108 and 209 cases were respectively selected in vancomycin and linezolid groups;108 cases with well-matched baseline characteristics were included in matched linezolid group.The clinical success rates of vancomycin vs.linezolid therapy were 86.1％ (93/108) and 88.9％ (96/108) (P =0.681),and the microbiologic success rates were 91.7％ (99/108) and 93.5％ (101/108) (P =0.795).The average trough concentration of vancomycin was (12 ± 8) mg/L.The rate of reaching the high trough concentration standard was only 33.3％ (36/108).In 50 (46.3％) cases the dose was adjusted according to the initial concentration data.Compared to less than 10 mg/L,the clinical efficacy of trough concentration in 10-20 mg/L was much higher(93.9％ (46/49) vs.78.6％ (33/42),P =0.031).Total bilirubin and platelet count had significant difference between the two-matched groups (34.1 (14.9,91.0)μmol/L vs.53.0(27.0,121.6) μmol/L,P =0.034;301.0 (198.8,416.0) × 109/L vs.195.5 (94.0,283.2) × 109/L,P =0.000).The incidence of linezolid related thrombocytopenia was 13.4％ (28 cases).Conclusion The clinical effect of vancomycin and linezolid on gram-positive bacterial sepsis in neonates is comparable.The rate of trough concentration of vancomycin reaching the high trough concentration standard is low,and the clinical efficacy is related to trough concentration.Linezolid have an effect on bilirubin and platelet count,the risk of thrombocytopenia should be monitored closely during linezolid treatment.

To explore the application of protein fingerprint technique and differential diagnosis in bacteriological negative pulmonary tuberculosis and pneumonia ,60 patients with bacteriological negative pulmonary tuberculosis ,60 patients with pneumonia ,and 60 healthy volunteers were selected from known clinical cases .Surface strengthening laser desorption ioniza-tion time of flight mass spectrometry (SELDI ToF Ms) and protein chip technology were applied to detect serum proteins ,and analyze their protein peaks by Ciphergen protein chip 3 .1 .1 software .Comparison of the serum protein fingerprinting data from the pool of 180 patients and healthy volunteers showed significant difference in 5 protein peaks (1 028 .49 ,4 796 .56 ,7 564 .77 , 8 048 .02 ,and 11 526 .75 m/z) identified between pulmonary tuberculosis and pneumonia (P<0 .01) .The total effective rate of the 5 protein peaks as a diagnosis model for differential diagnosis of bacteriological negative pulmonary tuberculosis and pneumonia was 84 .2% (101/120) ,the specificity was 82 .5% (52/63) ,the sensitivity was 85 .9% (49/57) ,the positive pre-dictive value was 86 .7% (52/60) ,and the negative predictive value was 81 .7% (49/60) .The total effective rate of the diagno-sis model for differential diagnosis of bacteriological negative pulmonary tuberculosis ,pneumonia and healthy volunteers was 89 .4% (161/180) .The specificity was 100% (60/60) ,the sensitivity was 84 .2% (101/120) ,the positive predictive value was 100% (101/101) ,and the negative predictive value was 75 .9% (60/79) .Protein fingerprinting technology is advanta-geous of being a simple method ,quick detection ,and requires less amount of sample .It is an effective means to screening the tuberculosis specific markers .We found the good diagnosis model through the detection of serum protein by protein fingerprint-ing technology .

Objective To explore the possible role of neuropilin-1 (NRP-1) and extracellular regulated protein kinases (ERK) in the mechanisms underlying the promotive effect of vascular endothelial growth factor (VEGF) on the proliferation of HaCaT cells.Methods HaCaT cells were cultured in vitro and transfected with a NRP-1 expression plasmid EX-O0008-M02.Reverse transcription (RT)-PCR and Western blot were performed to detect the mRNA and protein expression of NRP-1 in HaCaT cells respectively before and after the transfection.Some HaCaT cells were divided into three groups to be transfected with liposome (liposome control group),control plasmid pReceiver-M02 (plasmid control group),and objective plasmid EX-O0008-M02 (objective plasmid group),respectively,and each of the three groups was classified into several subgroups to be treated with phosphate buffer solution (PBS),U0126 (MEK1/2 inhibitor) and VEGF alone or in combination.After additional culture,methyl thiazolyl tetrazolium (MTT) assay was performed to determine the proliferative activity of HaCaT cells,Western blot to quantify the expression of total and phosphorylated ERK1/2 as well as proliferating cell nuclear antigen (PCNA) protein in HaCaT cells.Intergroup differences were assessed by one-way analysis of variance (ANOVA),and multiple comparisons and correction were done by using the least significant difference (LSD) test.Results RT-PCR and Western blot analysis confirmed that the transfection with NRP-1 effectively promoted the mRNA and protein expression of NRP-1 in HaCaT cells.A significant increase was observed in cellular proliferative activity (absorbence value at 570 nm) in the objective plasmid group compared with the liposome control group and plasmid control group (0.88 ± 0.14 vs.0.63 ± 0.07 and 0.62 ± 0.13,F =8.755,P ＜ 0.05),also in the VEGF-stimulated objective plasmid group compared with the VEGF-stimulated plasmid control group (1.14 ± 0.18 vs.0.88 ± 0.10,F =4.591,P ＜ 0.05).The U0126 pretreatment markedly suppressed the VEGF-induced proliferation of A375 cells in the objective plasmid group (0.50 ± 0.13 vs.1.14 ± 0.18,F =42.106,P ＜ 0.01).As Western blot analysis suggested,the objective plasmid significantly enhanced the VEGF-induced increase in ERK1/2 phosphorylation degree and PCNA expression intensity in HaCaT cells compared with the control plasmid,but the enhancing effect of objective plasmid was effectively inhibited by U0126.Conclusion The activation of ERK1/2 signaling pathway may play a key role in the NRP-1 protein-mediated promotive effect of VEGF on the proliferation of HaCaT cells.

Web impact of 30 foreign biomedical OA journal websites was evaluated using link analysis method.The current situation and existing problems of foreign medical OA Journal websites were discussed combined with evaluation results in order to provide references for the construction of medical OA journal websites in China.

Objective To compare the influence of open access journals ( OAJ ) of biomedicine on academic exchanges by empirically analyzing the journals enrolled in the Chinese science citation database-the core (CSCD-C).Methods Multivariate statistical analysis was performed on OAJ and non OAJ in CSCD by using bibliometric and statistical methods,SPSS software and rank-sum test.Results The OAJ were accounted for only 31.63％ of the total enrolled journals,the average ratio of funded papers in OAJ was 72.29％.Statistical journals had high influence in the discipline.The selfcitation rates of clinical medicine OAJ and special medical OAJ were high.The average impact factor and h index in biological OAJ were high.The average impact factor of preventive medicine OAJ was high.The average h index of comprehensive non OAJ was high.Conclusions The distribution of OAJ is uneven among different disciplines.The ratio of funded papers is higher in OAJ than in non OAJ with insignificant differences in discipline influence.

Objective To explore the sensitivity and the molecular mechanism of cisplatinresistance ovarian cancer cell line C13 to proteasome inhibitors and the combination with cisplatin. Methods After different treatments, methyl thiazolyl tetrazolium (MTT) assay was applied to examine the cell viability, annexin-V/propidium iodide(PI) apoptosis detection kit was used to determine the apoptosis rate of different groups, western blot assay was introduced to evaluate the expression levels of Fas-associated death domain-like interleukin-1 beta converting enzyme inhibitory protein (cFLIPs), and the activity of caspase-8 was examined. Results MTT assay shown that the cell viability ratios of combination group at serial time points from 12, 24, 36, 48, 60, 72 hours were ( 56.0 ± 8.4 ) %, (44.7 ± 7.3 ) %, ( 33.7 ±11.2) %, (27.6 ± 8.0) %, (27. 6 ± 7.6) % and (28.1 ± 2.4) %, which were much lower than those of cisplatin group (P <0.05). After treated for 24 hours, apoptosis rates of cisplatin group, bortezomib group and combination group were ( 16.7 ± 1.7) %, (23.4 ± 2.1 ) % and (26.9 ± 1.6) %, respectively. The rate of combination group was much higher than that of non-treated group and that of cisplatin group or bortezomib group ( P < 0.05 ). Western blot assay showed the changes of expression levels of cFLIPs, which were downregulated seriously after cisplatin, bortezomib or combination treatment [ (43.2 ± 2.3 )% vs( 75.7 ± 3.0)%vs (67.9 ± 2.1 ) %, P < 0.05 ]. The caspase-8 activity of combination group was (5.6 ± 1.6) folds than that of non-treated group, which was higher than those of other two groups [ ( 2.3 ± 1.0) and (4.2 ± 0.9 ) folds,P < 0.05 ]. Conclusions The tumor cell lethal effect of cisplatin could be increase significantly by the combination application of proteasome inhibitors, bortezomib. And the cFLIPs/caspase-8 signaling pathway may be play an important role in the molecular mechanism of the combination treatment.