Liver pathology is the gold standard for the diagnosis of chronic hepatitis.Although there are many effective noninvasive diagnostic methods,liver pathology is still the most direct and reliable method for evaluating inflammation and fibrosis in patients with chronic hepatitis.The classification system for liver pathology has been also constantly changing,so as to provide more accurate information of grading and staging and to guide clinical and scientific research.This article reviews the development of classification systems for liver pathology,the basic criteria and disadvantages of each major classification system,and the prospects of pathological classification systems for chronic hepatitis.

To begin with, we constructed cfp10-esat6 fusion gene and its prokaryotic expression vector and had it express in E. coli. By GeneSOEing techniques, a fusion gene was constructed by splicing cfpl0 gene and esat6 gene, and then was cloned into pGEX-4T-1 plasmid. Secondly, we constructed the prokaryotic expression recombinant plasmid pGcfp10-esat6. After identification with restriction enzyme analysis, PCR and nucleotide sequencing analysis, The E. coli BL21 containing the recombinant plasmid was induced by IPTG (Isopropy-beta-D-thiogalatoside). The fusion protein CFP10-ESAT6 with GST-tag about 42 kDa was expressed and purified with GST-fusion protein purification kit,The expression of cfp10-esat6 fusion gene was subsequently detected by SDS-polyacrylamine gel electrophoresis and Western-blot analysis. The sequence of cfp10 and esat6 in recombinant plasmid was consistent with that of GenBank report. The fusion protein existed in cytoplasm in soluble form and represented about 40% total bacterial protein of E. coil. The fusion protein was purified and the purity reached 90%. Its antigenicity was confirmed by Western-blotting. The prokaryotic expression vector (pGcfp1o-esat6) was constructed successfully, and the fusion protein CFP10-ESAT6 was obtained. This study provided an experimental basis for potential application of the recombinant CFP10-ESAT6 in the diagnosis of tuberculosis.
Antigens, Bacterial ; biosynthesis ; genetics ; Bacterial Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Mycobacterium tuberculosis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification

Background and objectiveRecent research shows thioridazine which is a kind of phenothiazine anti-psychotic drugs can inhibit the proliferation of various tumor cellsin vitro, but the role of thioridazine on lung cancer has not been reported. So we choose PC9 cell lines as the research object, the aim is to oberve the killing effect of thioridazine on PC9 cells and investigate its possible mechanism.MethodsAtfer being treated with different concentrations of thioridazine, the proliferation of PC9 cells was determined by methyl thiazolyltetrazolium (MTT) assay. Flow cytometry was used to measure the cell cycle distribution and apoptosis. The expressions of cell cycle-associated protein CyclinD1 and apoptosis-related proteins Caspase-3, Caspase-8, Caspase-9, Bcl-2, Bax and Bcl-xl were detected by Western blot.Results hTe proliferation of PC9 cells was signiifcantly inhibited by thioridazine in a dose- and time-dependent manner. Flow cytometry showed that cell cycle was arrested in G0/G1 phase and the apoptotic rates were signiifcantly increased with the increasing concentration of thioridazine. Compared with the control group, the differences were statistically signiifcant (P<0.05). Western blot analysis showed that, compared with the control group, thioridazine reduced the expressions of CyclinD1, Bcl-2 and Bcl-xl (P<0.01) and increased the expression of Bax (P<0.01). In the mean time, thioridazine promoted the activities of Caspase-3, Caspase-8 and Caspase-9 (P<0.01).ConclusionhTe mechanism of thioridazine inhibiting the proliferation of PC9 cells may be related to stimulation of Caspase apoptotic pathway, down-regulation of CyclinD1, Bcl-2, Bcl-xl and up-regulation of Bax.

Background and objective Tumor necrosis factor-related apoptosis-inducting ligand (TRAIL) can in-duce apoptosis of tumor cells, however, various of tumor cells may survive because of resistance to TRAIL-mediated apoptosis. This study is to observe the proliferation inhibition effect of TRAIL sensitized by thioridazine on PC9 cells through endoplas-mic reticulum (ER) stress mediated up-regulation of death receptor 5 (DR5) and investigate its mechanism.Methods PC9 cells were treated with different concentrations of thioridazine and TRAIL alone or in combination. Cell proliferation was mea-sured by MTT assay, and cell apoptosis and cell-surface DR5 were detected by flow cytometry. Western blotting was utilized to measure the expressions of ER stress-related proteins glucose regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), p-PKR-like ER kinase (PERK), p-eukaryotic initiation factor-2α (eIF2α), activating transcription factor 4 (ATF4) and apoptosis-related proteins caspase-3, caspase-9, caspase-8, PARP, DR5.Results Thioridazine inhibited the proliferation of PC9 cells in a dose-dependent manner (P<0.05). Thioridazine increased the inhibition and apoptosis of PC9 cells and up-regulated the expression of cell-surface DR5 induced by TRAIL. Flow cytometry showed that compared with TRAIL group,combination group of TRAIL and thioridazine increased cell apoptotic rates significantly (P<0.05). Western blotting indicated that compared with TRAIL group, expressions of Cleaved-caspase-8, Cleaved-PARP and DR5 increased significantly in combi-nation group of TRAIL and thioridazine. The induction of DR5 and pro-apoptotic effect were mediated through activation of ER stress accompanying by increased synthesis of GRP78 and CHOP, which can be blocked by adding of ER stress inhibitor 4-PBA.Conclusion Thioridazine enhanced proliferation inhibition effect of TRAIL in PC9 cells may be facilitated through ER stress mediated upregulation of DR5.

Genomic disorders caused by pathogenic copy number variation (pCNV) have proven to underlie a significant proportion of birth defects. With technological advance, improvement of bioinformatics analysis procedure, and accumulation of clinical data, non-invasive prenatal screening of pCNV (NIPS-pCNV) by high-throughput sequencing of maternal plasma cell-free DNA has been put to use in clinical settings. Specialized standards for clinical application of NIPS-pCNV are required. Based on the discussion, 10 pCNV-associated diseases with well-defined conditions and 5 common chromosomal aneuploidy syndromes are recommended as the target of screening in this consensus. Meanwhile, a standardized procedure for NIPS-pCNV is also provided, which may facilitate propagation of this technique in clinical settings.
Aneuploidy ; Cell-Free Nucleic Acids/genetics* ; Consensus ; DNA Copy Number Variations ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Pregnancy ; Prenatal Diagnosis

With the extensive application of highly sensitive genetic techniques in the field of prenatal diagnosis, prenatal chromosomal mosaicisms including true fetal mosaicisms and confined placental mosaicisms are frequently identified in clinical settings, and the diagnostic criteria and principle of genetic counseling and clinical management for such cases may vary significantly among healthcare centers across the country. This not only has brought challenges to laboratory technician, genetic counselor and fetal medicine doctor, but can also cause confusion and anxiety of the pregnant woman and their family members. In this regard, we have formulated a consensus over the prenatal diagnosis and genetic counseling for chromosomal mosaicisms with the aim to promote more accurate and rational evaluation for fetal chromosomal mosaicisms in prenatal clinics.
Consensus ; Female ; Genetic Counseling ; Humans ; Mosaicism ; Placenta ; Pregnancy ; Prenatal Diagnosis/methods*

Based on single-cell sequencing of the hippocampi of 5x familiar Alzheimer's disease(5x FAD)and wild type mice at 2-,12-,and 24-month of age,we found an increased percentage of microglia in aging and Alzheimer's disease(AD)mice.Blood brain barrier injury may also have contributed to this increase.Immune regulation by microglia plays a major role in the progression of aging and AD,according to the functions of 41 intersecting differentially expressed genes in microglia.Signaling crosstalk between C-C motif chemokine ligand(CCL)and major histocompatibility complex-1 bridges intercellular communi-cation in the hippocampus during aging and AD.The amyloid precursor protein(APP)and colony stimulating factor(CSF)signals drive 5x FAD to deviate from aging track to AD occurrence among intercellular communication in hippocampus.Microglia are involved in the progression of aging and AD can be divided into 10 functional types.The strength of the interaction among microglial subtypes weakened with aging,and the CCL and CSF signaling pathways were the fundamental bridge of communication among microglial subtypes.

Objective:To investigate the correlation between serum hypoxia inducible factor 1 alpha (HIF-1α), vascular endothelial growth factor (VEGF), soluble triggering receptor expressed on myeloid cells-1 (sTREM-1), and inflammatory factors and incision infection after anal fistula surgery.Methods:A retrospective analysis was conducted on the clinical data of 120 patients who underwent anal fistula thread hanging surgery at the First People′s Hospital of Shuangliu District, Chengdu from June 2022 to April 2023. The patients were divided into an infected group (36 cases) and a non infected group (84 cases) based on their postoperative incision infection status. The differences in serum HIF-1 α, VEGF, sTREM-1 and inflammatory factors [C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor α (TNF-α)] between the infected and non infected groups were compared, and the correlation between CRP, IL-6, TNF-α, HIF-1 α, VEGF, sTREM-1 and postoperative infection of anal fistula was analyzed by Pearson correlation coefficient; The correlation between CRP, IL-6, TNF-α and HIF-1 α, VEGF, sTREM-1. And the efficacy of serum HIF-1 α, VEGF, sTREM-1, inflammatory factors and their individual and combined diagnosis of incision infection after anal fistula surgery was analyzed by receiver operating characteristic (ROC) curve analysis.Results:The levels of serum HIF-1 α, sTREM-1, CRP, IL-6, and TNF-α in the infected group were higher than those before surgery and higher than those in the uninfected group 3 days after surgery; VEGF levels were lower than preoperative levels and lower than those in the non infected group (all P<0.05). Pearson correlation analysis showed that HIF-1α, sTREM-1, CRP, IL-6, TNF-α were positively correlated with postoperative infection in anal fistula ( r=0.456, 0.494, 0.455, 0.510, 0.363, all P<0.05), while VEGF was negatively correlated with postoperative infection in anal fistula ( r=-0.462, P<0.05). CRP, IL-6, TNF-α were positively correlated with HIF-1 α and sTREM-1 ( r=0.574/0.611/0.653, 0.661/0.608/0.610, all P<0.05), while CRP, IL-6, TNF-α were negatively correlated with VEGF ( r=-0.200, -0.207, -0.254, all P<0.05). The area under the curve (AUC) of HIF-1 α, VEGF, sTREM-1, CRP, IL-6, and TNF-α for diagnosing incision infection after anal fistula surgery were 0.716, 0.787, 0.741, 0.678, 0.792, and 0.688, respectively. The AUC of the combined diagnosis of inflammatory factors and 6 data points for postoperative incision infection in anal fistula surgery were 0.836 and 0.921, respectively. Conclusions:Serum levels of HIF-1 α, sTREM-1, CRP, IL-6, and TNF-α are abnormally high in patients with incision infection after anal fistula surgery, while VEGF is abnormally low in expression. HIF-1 α, VEGF, sTREM-1, and inflammatory factors can be used as effective indicators for clinical diagnosis of incision infection after anal fistula surgery, and their combined diagnostic value is better. HIF-1 α and sTREM-1 are positively correlated with inflammatory factors, while VEGF is negatively correlated with inflammatory factors.

Objective To evaluate the efficacy and safety of a domestic arterial thrombus aspiration catheter in treating acute arterial ischemic events in the experimental dogs,and to compare this catheter with Penumbra suction catheter.Methods Acute ischemic embolism model was established in the external carotid and renal arteries of experimental dogs,and the experimental dogs were randomly assigned to the study group and control group.The embolized blood vessels were treated with thrombectomy.Results A total of 12 experimental dogs were enrolled in this study,with 6 dogs in each group.All of the 12 experimental dogs were successfully modeled.In the study group and the control group,the cumulative success rates of thrombectomy were 92.9％and 66.7％respectively(P＞0.05),the incidences of intraoperative vascular dissection were 0％and 8.3％respectively(P＞0.05),and the incidences of vasospasm were 35.7％and 0.75％respectively(P＞0.05).Conclusion In treating thrombus-embolized blood vessels with mechanical thrombectomy in experimental dogs,no statistically significant differences in the efficacy and safety exist between using domestic arterial thrombus aspiration catheter and using Penumbra suction catheter.(J Intervent Radiol,2023,32:1207-1210)

Methanol has become an attractive substrate for the biomanufacturing industry due to its abundant supply and low cost. The biotransformation of methanol to value-added chemicals using microbial cell factories has the advantages of green process, mild conditions and diversified products. These advantages may expand the product chain based on methanol and alleviate the current problem of biomanufacturing, which is competing with people for food. Elucidating the pathways involving methanol oxidation, formaldehyde assimilation and dissimilation in different natural methylotrophs is essential for subsequent genetic engineering modification, and is more conducive to the construction of novel non-natural methylotrophs. This review discusses the current status of research on methanol metabolic pathways in methylotrophs, and presents recent advances and challenges in natural and synthetic methylotrophs and their applications in methanol bioconversion.
Humans ; Methanol/metabolism* ; Metabolic Engineering ; Metabolic Networks and Pathways ; Biotransformation