Objective Study on the inhibitory effect of gallnut extract extract on MRSA β-lactamase. Methods Determination of inhibitory effect of gallnut extract on MRSA3002 by TTC method. β-lactamase was repeated by freezing and thawing method . Synergistic effect of gallnut extract and gentamicin was detected by TTC. Results The MIC and MBC of MRSA3002 by gallnut extract were 8mg/mL and 32mg/mL.Gallnut extract can reduce strains of β-lactamase activity,the MRSA300224h 1/2MIC after the effect of gallnut extract, beta lactam enzyme activity inhibition compared with the control group there were significant differences (P<0.01),compared with the positive control group, the difference was not significant. Synergistic effect of gallnut extract and gentamicin can significantly reduce the MIC of MRSA3002. Conclusion Gallnut extract can reduce β-lactamase activity recovery sensitivity of drug-resistant bacteria.

Background:Zinc finger protein of cerebellum 2(ZIC2)is a transcriptional activator or repressor that is important for the organogenesis of the central nervous system.Previous studies have reported that ZIC2 is widely upregulated in a variety of tumors. Methods:Oncomine database was used to evaluate the expression levels of ZIC2 in hepatocellular car-cinoma(HCC)and normal liver tissues.Quantitative real-time polymerase chain reaction and immu-nohistochemistry were conducted to validate the results from the database.Cox regression analysis and survival curves were performed to assess the survival effect of ZIC2 in HCC. Results:Increased expression of ZIC2 was detected in HCC tissues compared with normal liver tissues.In addition,patients with high ZIC2 levels had a poor prognosis.Multivariate analysis showed that clinical stage(T or M classification)and ZIC2 levels were independent prognostic factors for overall survival.Moreover,a subgroup analysis revealed that patients with moderate or poor grade tumors,T1-2 tumors,N0 tumors,early American Joint Committee on Cancer(AJCC)stage,and old age were more likely to present with overexpression of ZIC2.To conclude,ZIC2 is upregulated in HCC and associated with the histology and survival of HCC patients. Conclusions:The expression status of ZIC2 may serve as an independent prognostic marker of HCC.

BACKGROUND:It is reported that bone marrow mesenchymal stem cell(BMSC)transplantation might be a promising treatment for liver fibrosis.But the mechanism is still unclear.OBJECTIVE:To observe the hepatic stellate cells apoptosis induced by BMSC transplantation,and to study the mechanism of BMSC in treating hepatic fibrosis in vivo.METHODS:CCl_4 subcutaneous injection was performed to induce rat liver fibrosis.After 8 weeks of CCU injection,20 rats which underwent successful model establishment were randomly divided into experimental group and control group,10 in each group.The experimental group received MSC transplantation via tail vein injection,and the control group were given DMEM instead.The rats were killed and the livers were harvested at three time point,the day of MSC transplantation,3 days after transplantation,and 7 days after transplantation.The hydroxyproline content was detected by HE and Masson staining,and the expression changes of α-smooth muscle actin(α-SMA)proteins were determined using immunohistochemistry.The apoptosis of hepatic stellate cells were determined by α-SMA and TUNEL(terminal dUTP nick-end labeling)dual-staining.RESULTS AND CONCLUSION:After 8 weeks of CCU injection,the hydroxyproline content increased and histology indicated progress of liver fibrosis.At 7 days after MSC transplantation,the hydroxyproline in the liver was decreased,and the liver fibrosis was alleviated in the experimental group but aggravated in the control group.Immunohistochemistry indicated that α-SMA positive cells were increased at 8 weeks after CCU injection.At day 7 after transplantation,α-SMA positive cells in the experimental group were significantly less than control group(P ＜ 0.05).At 3 days after transplantation,the hepatic stellate cells apoptosis in the experimental group was significantly aggravated compared with control group(P ＜ 0.05).This suggested that MSC transplant was an effective treatment for liver fibrosis.MSC inducing hepatic stellate cells apoptosis may be one of the mechanisms.

BACKGROUND:Previous studies have confirmed that bone marrow mesenchymal stem cells in vitro can promote hepatic stel ate cellapoptosis and inhibit its activity, in which the mechanism of action remains unknown. OBJECTIVE:To screen out apoptosis-related genes during hepatic stel ate cellapoptosis regulated by bone marrow mesenchymal stem cells using gene chip technology. METHODS:Purified human bone marrow mesenchymal stem cells were seeded in 6-wel Transwel plate and cocultured with hepatic stel ate cells. Cultured human bone marrow mesenchymal stem cells alone served as control group, and cultured for 72 hours. The alterations in apoptosis-related genes were analyzed between culture alone group and coculture group using gene chip technology. The genes strongly associated with regulation of hepatic stel ate cells were selected. RESULTS AND CONCLUSION:By the functional classification of second-generation SABiosciences Gene chips, apoptotic gene screening found that after coculture, significantly upregulated genes in bone marrow mesenchymal stem cells contained:AKT1, PIK3R2, DAPK1, DHCR24, NOTCH2 and BDNF. Combined with previous findings, we hypothesized that NOTCH may play a key role in the regulation of hepatic stel ate cells by bone marrow mesenchymal stem cells.

Objective To observe the reproducibility of radiomics feature(RF)extracted from virtual monoenergetic image(VMI)of rabbit VX2 hepatoma models obtained with 3 different dual-energy CT(DECT)systems,and to explore relationship of reproducibility and diagnostic performance of RF.Methods Fifteen rabbits with VX2 hepatoma were randomly divided into 3 groups(each n=5).Contrast-enhanced abdominal CT scanning under volume CT dose index(CTDIvol)levels of 6,9 and 12 mGy were performed with dual-source DECT(dsDECT),rapid kV switching DECT(rsDECT)and dual-layer detector DECT(dlDECT),respectively.VMI were reconstructed at 10 keV increments from 40 to 140 keV.RF were extracted from VMI,the reproducibility was assessed using intra-class correlation coefficient(ICC),and those with ICC≥0.8 were considered as reproducible RF.The percentage of reproducible features(denoted by R)were compared among different scanner pairings and different CTDIvol levels.Within each CTDIvol group,the reconstruction energy levels yielding the maximum number(denoted by N)of common RF across different scanner pairings were identified.The receiver operating characteristic(ROC)curve was drawn,the area under the curve(AUC)was calculated,and the diagnostic efficacies of reproducible RF and other RF were compared under optimal reproducible conditions.Spearman correlation coefficient between ICC and the corresponding AUC of RF were calculated.Results RrsDECT-dsDECT(6.45％,95％CI[2.36％,8.87％])was higher than RdlDECT-dsDECT(0.72％,95％CI[0.15％,1.79％])and RrsDECT-dlDECT(1.43％,95％CI[0.60％,4.06％])(all adjusted P＜0.05),R9mGy(3.70％,95％CI[1.31％,5.73％])and R12mGy(2.63％,95％CI[0.60％,6.69％])were higher than R6mGy(1.31％,95％CI[0.12％,1.55％])(all adjusted P＜0.05).The optimal reproducible reconstruction energy levels of RF under CTDIvol of 6,9 and 12 mGy concentrated at 50-70 keV.AUC of reproducible RFs were higher than of other RF(all adjusted P＜0.05)and had certain correlation with the reproducibility(rs=0.102-0.516,P＜0.05).Conclusion The reproducibility of RF extracted from contrast-enhanced VMI CT images of rabbit VX2 hepatoma models associated with DECT scanner,CTDIvol level and reconstruction energy level.RF with higher reproducibility might have better diagnostic performance.

Objective:To report the BEST1 gene mutations and clinical phenotypes in two pedigrees with Best vitelliform macular dystrophy (BVMD) and autosomal recessive bestrophinopathy (ARB). Methods:A retrospective clinical study. From November 2019 to March 2021, in the Department of Ophthalmology of The First Affiliated Hospital of Zhengzhou University, the BVMD family (4 patients and 6 family members) and the ARB family (2 patients, 2 family members), a total of 6 patients and 8 normal family members were included in the study. Detailed medical history was obtained; best corrected visual acuity, fundus color photography, electrophysiology, optical coherence tomography and fundus autofluorescence examination were performed. The clinical characteristics for all patients in the two families were analyzed. Three milliliter peripheral venous blood of all participants in the family was collected, and the whole genomic DNA was extracted with gene sequencing using next-generation sequencing technology based on targeted capture. Compared with the database to identify the pathogenicity mutation sites, suspected pathogenic mutation sites were selected, then mutations in other members in the family was assayed by Sanger sequencing.Results:In family 1, the proband was demonstrated as typical BVMD, other patients were multifocal vitelliform macular dystrophy. The DNA sequencing result showed that all the 4 patients carried heterozygous missense mutations in exon 3 of BEST1 gene: c.240C>G (p.F80L) (M1) and 2 members carried this mutation, but without clinical phenotype. M1 was a likely-pathogenic mutation reported for the first time. In family 2, the proband and the other patient were diagnosed as ARB. The DNA result showed that the 2 patients carried heterozygous missense mutations in exon 5 and exon 2 of BEST1 gene: c.584C>T (p.A195V) (M2)、c.139C>A (p.R47S) (M3), and a heterozygous frameshift mutation in exon 3 of BEST1 gene: c.235dupT (p.S79Ffs*153) (M4). M2 was a pathogenic mutation reported previously. M3 variant was of undetermined significance. M4 was a first reported pathogenic mutation. Conclusions:The BEST1 gene mutation is the main cause of BVMD and ARB. Different mutation sites have different clinical phenotypes. BVMD and ARB have genetic and clinical heterogeneity.

Objective Sequencing depth is a key parameter in next generation sequencing,which is closely related to the accuracy of sequencing results.Forensic biological evidence examination requires extremely high accuracy.It is crucial to scientifically and reasonably set the sequencing depth analysis threshold for forensic next generation sequencing testing.Methods This study used targeted sequencing data of microhaplotypes from 50 samples with known genotypes.By calculating the accuracy,precision,recall,and F1 score of each locus under various threshold conditions,two types of analysis threshold setting methods,which were based on fixed read count and fixed sequencing depth ratio,were studied extensively.Results The results showed that false positives were observed when the analysis threshold was set at 50×or 100×.When the analysis threshold was set at 200×,false negatives were observed.When the analysis threshold was set at 1.5%,3.0%,or 4.5%,false positives were observed.This study further proposed a third type of analysis threshold setting method,which was based on sequencing depth ratio scatter plots.With this method,no false positive or false negative was observed in the results.This article then explored four factors that lead to significant differences in the sequencing depth of forensic next generation sequencing experiments,compared with the analysis threshold setting method for capillary electrophoresis technology,and discussed the correlation between analysis thresholds and the ability to distinguish mixed DNA.Conclusion Employing the sequencing depth ratio scatter plot method to set analysis threshold has significant application value in next generation sequencing-based forensic genetic marker genotyping.