Ferroptosis and drug-induced liver injury / 中华肝脏病杂志

Ferroptosis is a type of regulated cell death driven by iron-dependent lipid peroxidation that has received extensive attention in recent years. A growing body of evidence suggests that ferroptosis contributes to the progression of drug-induced liver injury. Therefore, the role and mechanism of ferroptosis in the process of drug-induced liver injury deserve further extensive and in-depth exploration, which will aid in the discovery of novel biomarkers as well as the identification of potential approches of targeting ferroptosis to intervene in drug-induced liver injury.

Active constituents of Urtica fissa in inhibition of benign prostatic hyperplasia / 中国中药杂志

The present study investigated the material basis of Urtica fissa for the inhibition of benign prostatic hyperplasia(BPH). The active fractions were screened, and the extracts of dichloromethane and ethyl acetate exhibited significantly inhibitory activities against 5α-reductase in vitro and BPH in model rats. The chemical constituents in the active fractions were systematically investigated, and 28 compounds were obtained, which were identified as lobechine methyl ester(1), dibutyl-O-phthalate(2), 1-monolinolein(3), epipinoresinol(4), 5-hydroxy-3,4-dimethyl-5-pentanyl-2(5H)-furanone(5), E-7,9-diene-11-methenyl palmitic acid(6), evofolin B(7), ficusal(8), threo-2,3-bis-(4-hydroxy-3-methoxyphenyl)-3-ethoxypropan-1-ol(9), α-viniferin(10),(9R,7E)-9-hydroxy-5,7-mengatigmadien-4-one-9-O-β-D-glucopyranoside(11), indole-3-carboxaldehyde(12), p-hydroxy ethyl cinnamate(13), benzyl alcohol-O-β-D-glucoside(14), L-methionine(15), 4-methoxyaniline(16), 6-aminopurine(17), 8'-acetyl oilvil(18), 4-methoxyl-8'-acetyl oilvil(19), vanillic acid(20), β-hydroxypropiovanillone(21), 7-hydroxy-6-methoxycoumarin(22), p-hydroxybenzaldehyde(23), pinoresinol(24), erythro-1,2-bis-(4-hydroxy-3-methoxyphenyl)-1,3-propanediol(25), urticol(26), urticol-7-O-β-D-glucopyranoside(27), and lobechine(28). Compounds 1-17 were isolated from U. fissa for the first time. Meanwhile, compound 1 was a new natural product. Compounds 10, 11, 19, 21, and 27 exhibited significant inhibitory effects on 5α-reductase.

MicroRNA-141-3p expression in serum of patients with cerebral hemorrhage and its mechanism / 解剖学报

Objective To analyze the expression level of microRNA-141-3p (miR-141-3) in patients with intracerebral hemorrhage (ICH), and explore the effect and mechanism of miR-141-3p on cerebral hemorrhage injury in rats. Methods Forty patients with ICH and 40 healthy controls in total were enrolled in this study. The expression of miR- 141-3p in peripheral blood serum was determined by the Real-time PCR method. The target relationship between miR-141- 3p and NOD-like receptor 3 (NLRP3) 3′ UTR was confirmed by dual luciferase reporter assay. miR-141-3p agonist and agonist NC were injected into rats via the lateral ventricle, respectively. On day 7 after treatment, the neurological function score was evaluated, and then all rats were killed to obtain brain tissue. Brain water content was examined by the dried and wet mass. HE staining was conducted to observe the pathological changes of cerebral tissue. The mRNA expressions of NLRP3 and miR-141-3p were detected by Real-time PCR. The protein expression of interleukin (IL)-lβ, IL-6 and IL-18 were detected by Western blotting analysis. Results The expression of miR-141-3p in serum of ICH patients was significantly down-regulated compared to healthy controls and negatively correlated with the severity of edema around the hematoma [(0.068±0.038) vs (0.520±0.028), t = 15.93, P<0.001; r =-0.8948, -0.9434 to-0.8087, P<0.001 ]. The result of luciferase reporter assay showed that miR-141-3p was related to the regulation of NLRP3 gene expression. The relative expression levels of miR-141-3p in agonist group were significantly higher than those in the agonist NC group (P< 0.001), while the expression levels of NLRP3, IL-lβ, IL-6 and IL-18 were significantly lower than those in the agonist NC group (P< 0.001). Compared with the agonist NC group, the cerebral water content reduced significantly (P< 0.001), and the neurological function score was significantly improved on the day 7 after treatment in agonist group (P< 0.001). The result of HE staining showed that injection of miR-141-3p in ICH rats could reduced the severity of edema around the hematoma. Conclusion MiR-141-3p alleviates ICH-induced inflammatory injury in rat possibly by modulating miR-141-3p.

Lignans with inhibitory effect on 5α-reductase from Urtica cannabina / 中国中药杂志

The lignans in Urtica cannabina were isolated by preparative HPLC, silica, and ODS column chromatographies, and identified by NMR and HR-MS. The inhibitory activities on 5α-reductase were evaluated in vitro. As a result, ten secolignans,(2R,4S)-2,4-bis(3-methoxyl-4-hydroxyphenyl)-3-butoxypropanol(1), 3,4-trans-3-hydroxymethyl-4-[bis(3,4-dimethoxyphenyl)methyl] butyrolactone(2), 3,4-trans-3-hydroxymethyl-4-[(3,4-dimethoxyphenyl)(3-methoxyl-4-hydroxyphenyl)methyl] butyrolactone(3), 3,4-trans-3-hydroxymethyl-4-[bis(3-methoxyl-4-hydroxyphenyl)methyl] butyrolactone(trans urticol, 4), 3,4-trans-3-hydroxymethyl-4-[bis(3,4-dimethoxyphenyl)methyl] butyrolactone-3-O-β-D-glucopyranoside(5), 3,4-trans-3-hydroxymethyl-4-[(3,4-dimethoxyphenyl)(3-methoxyl-4-hydroxyphenyl)methyl]butyrolactone-3-O-β-D-glucopyranoside(6), 3,4-trans-3-hydroxymethyl-4-[bis(3-methoxyl-4-hydroxyphenyl)methyl]butyrolactone-3-O-β-D-glucopyranoside(trans-urticol-7-O-β-D-glucopyranoside, 7), cycloolivil-4-O-β-D-glucopyranoside(8), isolariciresinol-4'-O-β-D-glucopyranoside(9), and olivil-4'-O-β-D-glucopyranoside(10), together with a polyphenol [α-viniferin(11)], were isolated from U. cannabina for the first time. Compound 1 was a new lignan. Compound 7 was potent in inhibiting 5α-reductase.

Natural products as potent inhibitors of hypoxia-inducible factor-1α in cancer therapy / 中国天然药物

Hypoxia is a prominent feature of tumors. Hypoxia-inducible factor-1α (HIF-1α), a major subunit of HIF-1, is overexpressed in hypoxic tumor tissues and activates the transcription of many oncogenes. Accumulating evidence has demonstrated that HIF-1α promotes tumor angiogenesis, metastasis, metabolism, and immune evasion. Natural products are an important source of antitumor drugs and numerous studies have highlighted the crucial role of these agents in modulating HIF-1α. The present review describes the role of HIF-1α in tumor progression, summarizes natural products used as HIF-1α inhibitors, and discusses the potential of developing natural products as HIF-1α inhibitors for the treatment of cancer.

Regulatory Effect of Sijunzi Tang and Its Single Herbs on Intestinal Flora in Rats with Spleen Deficiency / 中国实验方剂学杂志

Objective:To investigate the regulatory effect of Sijunzi Tang(SJZT) and its single herbs(Ginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma Praeparata cum Melle, Atractylodis Macrocephalae Rhizoma and Poria) on intestinal flora in spleen-deficient rats. Method:Normal rats were randomly divided into the blank group, model group, Zhengchangsheng granules group, SJZT group and each single herb group, rats were orally administered Sennae Folium decoction to induce diarrhea for ten consecutive days to establish a spleen-deficient model(distilled water for the blank group), then treated with the corresponding drugs for seven consecutive days(distilled water for the blank group and the model group). Fresh feces were collected on pre-modeling(0th day), post-modeling(11th day), and post-treatment(18th day). Short-chain fatty acids(SCFAs) in feces were acidified by sulphuric acid and extracted by diethyl ether, then determined by gas chromatography. The structural change(diversity and similarity) of intestinal flora in feces was analyzed by 16S rDNA-polymerase chain reaction(PCR)-denaturing gel gradient electrophoresis(DGGE) technique. Result:Compared with blank group, the contents of SCFAs as well as diversity and similarity indexes of intestinal flora in feces of all administered groups were significantly decreased on the 11th day(PPth day, compared with model group, the contents of SCFAs as well as diversity and similarity indexes of intestinal flora in feces of Atractylodis Macrocephalae Rhizoma group were significantly increased(PPPPConclusion:Ginseng Radix et Rhizoma, Atractylodis Macrocephalae Rhizoma and Poria are the single herbs responsible for the regulatory effect of SJZT on intestinal flora in spleen-deficient rats, and Atractylodis Macrocephalae Rhizoma may play a major role.

Comprehensive mass spectrum analysis of two flavone-6,8-C-di-glycosides and its application by high resolution electrospray ionization tandem mass spectroscopy in both negative and positive ion modes / 中国中药杂志

The tandem mass spectrum of apigenin-6,8-C-di-glucoside( 1) and apigenin-6-C-glucose-8-C-rhamnoside( 2) were obtained by high resolution electrospray ionization mass spectrometry( HR-ESI-MS/MS) in both positive and negative ion modes. The elemental composition of each ion was determined according to its accurate mass-to-charge,hence,the fragmentation pathways of each compound were proposed in both negative and positive ion modes. Comprehensive analysis of each ion and its proposed fragmentation pathways of the two compounds was initially conducted in both negative and positive ion mode HR-ESI-MS/MS to explore the diagnostic ions for flavone-6,8-C-di-glycosides and the characteristic ions for each compound and their cleavage rules. The results showed that a family of fragmentation ions with m/z 353,325,311,297 in ESI(-)-MS and m/z 355,325,307,295 in ESI( +)-MS could be the diagnostic ions of flavone-6,8-C-di-glycoside,and characteristic neutral loss could be assigned to glycosyl substitution,for example,neutral losses of C_4H_8O_4( 120),C_3H_6O_3( 90),C_2H_4O_2( 60) for glucoside substitution while neutral losses of C_4H_8O_3(104),C_3H_6O_2( 74),C_2H_4O( 44) for rhamnoside substitution. Furthermore,only one H_2O loss from mother ion( [M-H]-) was observed for 1 & 2 in ESI(-)-MS while five to six H2 O loss from mother ion( [M+H]+) was observed for 1 & 2 in ESI( +)-MS to produce a family of ions by subsequent loss of H_2O,which could be applied for glucosyl difference. The flavone-6,8-C-di-glycosides in both ESI( +)-MS and ESI(-)-MS showed the cleavage similarity at sugar substitutions. However,there were much more differences by the fragmentation pathways and neutral losses between ESI( +)-MS and ESI(-)-MS as following,hyperconjugation ions by subsequent loss of H_2O from precursor ions of flavone-6,8-C-di-glycosides in ESI( +)-MS were not observed in ESI(-)-MS; the subsequent neutral loss of CH_2O in ESI( +)-MS were rarely observed in ESI(-)-MS; the loss of CO only happen at C-ring of flavone ESI( +)-MS other than glycosyl position in ESI(-)-MS; the C4-chain neutral loss of flavone-6,8-C-di-glycosides happened at 8-C-glycosyl position other than at 6-C-glycosyl position. The above cleavage rules and diagnostic ions of ESI( +)-MS were successfully applied for the structure identification of 4 flavone-6 C,8 C-diglycosides from the stem extract of Dendrobium officinale as vicenin Ⅱ,vicenin Ⅰ,isoschaftoside,schaftoside as well as one flavone-O-glysoside named rutin,which were supported by ESI(-)-MS data as well.

Studies on liposoluble constituents from Momordicae Semen / 中国中药杂志

The liposoluble constituents in Momordicae Semen were investigated in the present study. By silica gel, Sephadex LH-20 column chromatography, and semi-preparative HPLC, 22 compounds were isolated and purified from dichloromethane and ethyl acetate fraction. Based on NMR and MS spectra analyses, these compounds were identified as lupeol (1), 5-(1'-hydroxypentyl)-5H-furan-2-one (2), palmitic acid (3), viscumamide (4), clavatustide C (5), laxanol (6), threo-1-(4-hydroxyphenyl)-2-{4-[2-formyl-(E)-vinyl]-2-methoxyphenoxyl}-propane-1, 3-diol (7), α-spinasterol-3-O-β-D-glucoside (8), chushizisin F (9), ehletianol C (10), tanegool (11), (7R, 8R, 8'R)-4'-guaiacylglyceryl-evofolin B (12), ligballinone (13), (7R, 8S, 8'R)- 4, 4', 9-trihydroxy- 7, 9'-epoxy- 8, 8'-lignan (14), chushizisin I (15), chushizisin A (16), chushizisin G (17), p-coumaraldehyde (18), α-spinasterol (19), p-hydroxybenzoic acid (20), chushizisin E (21), and 3-[2-(4-hydroxyphenyl)-3-hydroxyphenyl-2, 3-dihydro-1-benzofuran-5-yl] propane-1-ol (22), respectively. Compounds 1-17 were isolated from Momordica cochinchinensis for the first time. Compound 2 was a new natural product while compounds 4 and 5 were first found in the terrestrial organism.

Screening for, andVariants in a Cohort of Chinese Patients with Charcot-Marie-Tooth / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>SH3TC2, PMP2, and BSCL2 genes are related to autosomal recessive (AR) Charcot-Marie-Tooth (CMT) disease type 1, autosomal dominant (AD)-CMT1, and AD-CMT2, respectively. Pathogenic variants in these three genes were not well documented in Chinese CMT patients. Therefore, this study aims to detect SH3TC2, PMP2, and BSCL2 pathogenic variants in a cohort of 315 unrelated Chinese CMT families.</p><p><b>METHODS</b>A total of 315 probands from 315 unrelated Chinese CMT families were recruited from the Department of Neurology of Third Xiangya Hospital and Xiangya Hospital. We screened for SH3TC2 pathogenic variants in 84 AR or sporadic CMT probands, PMP2 pathogenic variants in 39 AD or sporadic CMT1 probands, and BSCL2 pathogenic variants in 50 AD or sporadic CMT2 probands, using polymerase chain reaction and Sanger sequencing. All these patients were out of 315 unrelated Chinese CMT families and genetically undiagnosed after exclusion of pathogenic variants of PMP22, MFN2, MPZ, GJB1, GDAP1, HSPB1, HSPB8, EGR2, NEFL, and RAB7. Candidate variants were analyzed based on the standards and guidelines of American College of Medical Genetics and Genomics (ACMG). Clinical features were reevaluated.</p><p><b>RESULTS</b>We identified three novel heterozygous variants such as p.L95V (c.283C>G), p.L1048P (c.3143T>C), and p.V1105M (c.3313G>A) of SH3TC2 gene and no pathogenic variants of PMP2 and BSCL2 genes. Although evaluation in silico and screening in the healthy control revealed that the three SH3TC2 variants were likely pathogenic, no second allele variants were discovered. According to the standards and guidelines of ACMG, the heterozygous SH3TC2 variants such as p.L95V, p.L1048P, and p.V1105M were considered to be of uncertain significance.</p><p><b>CONCLUSIONS</b>SH3TC2, PMP2, and BSCL2 pathogenic variants might be rare in Chinese CMT patients. Further studies to confirm our findings are needed.</p>

Several emerging infectious diseases existing risks to China at present / 中国人兽共患病学报

In the past several years,more and more outbreaks of new infectious diseases emerged all over the world and some epidemics seriously affected China.Some kinds of zoonotic pathogens such as West Nile virus,Marburg virus,Lassa virus,Venezuelan equine encephalitis virus,Nipa virus,Trypanosoma brucei,Rift valley fever virus have caused seriously epidemics between human beings and animals.With the great development of the economy and international trade in China,the global spread of infectious diseases should cause great concern in china.

Impact of Qi-invigorating traditional Chinese medicines on intestinal flora: A basis for rational choice of prebiotics / 中国天然药物

According to the theory of traditional Chinese medicine (TCM), Qi (vital energy) is regarded as a driving force of biological activities in human body, including both nutrient substances and organ functions. Qi-invigorating TCMs are widely used to treat various symptoms and disorders, such as fatigue, obesity, immunosuppression, intestinal flora imbalance, and gastrointestinal diseases, in which Qi is considered to be reduced or depleted. Interestingly, abundant clinical evidences suggest that these disorders are associated with the alternation of intestinal flora, which directly affects disease status. Herein we review the interaction between gut microbiota and Qi-invigorating TCMs under healthy and disease conditions and discuss the mechanisms of action and applications of Qi-invigorating TCMs in enhancing health status through microbial alternation. A better understanding of the role of Qi-invigorating TCMs in modulating microbial composition and the association between intestinal microbiota and diseases would help reveal the clinical consequences of microbiota alteration and explore opportunities to harness this symbiotic relationship to improve public health.

Mechanisms of PPARα,sterol regulatory element binding proteins and AMP-activated protein kinase in alcoholic fatty liver diseases:review / 中国药理学与毒理学杂志

Alcoholic fatty liver disease (AFLD) is the first step to wards alcoholic liver disease (ALD). A beffer knowledge of AFLD will contribute to the prevention and therapy of ALD. It has been found that the occurrence and development of AFLD mainly involve the pathways of cytochrome P450 (CYP2E1),peroxisome proliferator-activated receptor α(PPARα),sterol regulatory element-binding proteins (SREBPs),AMP-activated protein kinase(AMPK),sirtuin 1(SIRT1),adiponectin and insulin.This review focuses on the importance of PPARα,SREBPs and AMPK pathway in alcoholic steatosis.It's reported that alcohol and its metabolite acetaldehyde inhibit PPARα and AMPK,and activate SREBP protein directly or indirectly,leading to liver lipid metabolic disorders,reducing the ability of fatty acid oxidation,causing lipid accumulation, and eventually inducing AFLD. Additionally, this review outlines the prospect of a therapeutics of AFLD targetting PPARα,SREBPs and AMPK.

Advance in studies on effect of Glycyrrhizae Radix et Rhizoma in relieving purgative activity of Rhei Radix et Rhizoma / 中国中药杂志

Rhei Radix et Rhizoma, as a commonly used traditional Chinese medicine, has been widely applied in clinic. Its major purgative constituent is anthraquinones, which are believed to be a toxic ingredient. Glycyrrhizae Radix et Rhizoma has been reputed as the best alexipharmic to moderate medicine natures. In this paper, the effect of Glycyrrhizae Radix et Rhizoma in relieving purgative activity of Rhei Radix et Rhizoma was studied in two aspects--the boiling process and intestinal metabolism; Studies on combined administration of Glycyrrhizae Radix et Rhizoma and Rhei Radix et Rhizoma in recent years were summarized according to chemical constituent, intestinal flora, I/II phase metabolism and drug transport. However, the material basis and mechanism for their compatibility remain unclear, further studies will be made in the future.

Quality standard study on Entadae Semen / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the quality standard of Entadae Semen, and provide scientific basis for its quality control.</p><p><b>METHOD</b>TLC and HPLC were used for qualitative identification and quantitative analysis of phaseoloidin and entadamide A-O-β-D-glucopyranoside in Entadae Semen. The test of water content, ash and ethanol-soluble extractives of Entadae Semen was carried out according to the methods recorded in appendix of Chinese Pharmacopeia (2010 edition).</p><p><b>RESULT</b>The TLC was well separated with clear spots. The linear range of phaseoloidin was between 0.014 to 2.747 g x L(-1) (r = 0.999 6, n = 9) with an average recovery rate of 101.06% (RSD 0.90%, n = 6); the linear range of entadamide A-O-β-D-glucopyranoside was between 0.002 to 0.452 g x L(-1) (r = 0.999 7, n = 9) withan average recovery rate of 101.52% (RSD 1.09%, n = 6). The content of phaseoloidin in sample is between 5.12% to 9.24%, entadamide A-O-β-D-glucopyranoside is between 0.55% to 2.17%, alcohol-soluble extracts is between 30.9% to 45.2%, water is between 6.6% to 8.6%, and total ash is between 2.4% to 2.9%.</p><p><b>CONCLUSION</b>The established standard is acceptable for quality control of Entadae Semen.</p>

Isolation and characterization of an algicidal bacterium indigenous to lake Taihu with a red pigment able to lyse microcystis aeruginosa / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To isolate and characterize indigenous algicidal bacteria and their algae-lysing compounds active against Microcystis aeruginosa, strains TH1, TH2, and FACHB 905.</p><p><b>METHODS</b>The bacteria were identified using the Biolog automated microbial identification system and 16S rDNA sequence analysis. The algae-lysing compounds were isolated and purified by silica gel column chromatography and reverse-phase high performance liquid chromatography. Their structures were confirmed by Nuclear Magnetic Resonance (NMR) and Fourier Transform Infrared (FT-IR) spectroscopy. Algae-lysing activity was observed using microscopy.</p><p><b>RESULTS</b>The algae-lysing bacterium LTH-2 isolated from Lake Taihu was identified as Serratia marcescens. Strain LTH-2 secreted a red pigment identified as prodigiosin (C20H25N3O), which showed strong lytic activity with algal strains M. aeruginosa TH1, TH2, and FACHB 905 in a concentration-dependent manner. The 50% inhibitory concentration (IC50) of prodigiosin with the algal strains was 4.8 (± 0.4)× 10⁻² μg/mL, 8.9 (± 1.1)× 10⁻² μg/mL, and 1.7 (± 0.1)× 10⁻¹ μg/mL in 24 h, respectively.</p><p><b>CONCLUSION</b>The bacterium LTH-2 and its pigment had strong Microcystis-lysing activity probably related to damage of cell membranes. The bacterium LTH-2 and its red pigment are potentially useful for regulating blooms of harmful M. aeruginosa.</p>

Simultaneous determination of chlorogenic acid, scutellarin, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid in different parts of Erigeron breviscapus by high-performance liquid chromatography / 中国中药杂志

<p><b>OBJECTIVE</b>A high-performance liquid chromatographic (HPLC) method was developed for simultaneous determination of chlorogenic acid, scutellarin, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid in different parts of Erigerontis Herba.</p><p><b>METHOD</b>The four constituents were measured on an Agilent Zorbax SB-C18 column (4.6 mm x 450 mm, 5 microm) with a gradient elution of acetonitrile (A) -0.3% phosphoric acid solution (B) (0-10 min, 12%-15% A, 10-32 min, 15% A, 32-33 min, 15%-20% A, 33-50 min, 20%-22% A) at wavelength of 335 nm and 327 nm, and a flow rate of 1.0 mL x min(-1) and the column temperature was 30 degrees C.</p><p><b>RESULT</b>Linearity of each standard was established in the concentration range of 0.050 1-1.002 microg for chlorogenic acid, 0.165 9-3.318 microg for chlorogenic acid, 0.049 7-0.994 microg for 3,5-dicaffeoylquinic acid, 0.048 7-0.974 p.g for 4,5-dicaffeoylquinic acid respectively, with correlation coefficient r > 0.999 6. Average recoveries (n = 6) of 4 compounds were 98.53% with a RSD of 0.94%, 99.68% with a RSD of 0.49%, 98.78% with a RSD of 1.1%, 99.06% with a RSD of 0.81%, respectively.</p><p><b>CONCLUSION</b>The developed method is simple, accurate, and precise, it can be used for the quantitative analysis of Erigeron breviscapus.</p>

Detection of antigen-receptor gene rearrangement clonality for diagnosis of lymphoma / 中国实验血液学杂志

This study was aimed to investigate the significance of detecting the antigen-receptor gene rearrangement clonality in the diagnosis of lymphoma. Paraffin-embedding and HE staining of samples from 31 patients with lymphomas were performed for morphologic observation by light microscope. Immunophenotype was analyzed by the immunohistochemistry (IHC) method. The clonality of antigen-receptor gene rearrangement was detected by BIOMED-2 Assay Kit. The results showed that among the 31 cases, 12 cases were suspected to be T-cell lymphoma, 1 case was suspected to be T-cell reactive hyperplasia, and 16 cases were suspected to be B-cell lymphoma, 2 cases were B-cell reactive hyperplasia. The detection results showed that the positivity of Ig gene rearrangement clonality was 94.44% (17/18), the positivity of TCR gene rearrangement clonality was 92.31% (12/13), the other two cases were negative. Finally, 12 cases were diagnosed to be T-cell lymphoma and 17 cases were B-cell lymphoma. The other two cases were reactive lymphoid proliferations. And the positivity rate in the 31 patients with lymphomas was 93%. It is concluded that the detection of antigen-receptor gene rearrangement clonality is a useful assistant method in the diagnosis of lymphoma.

Cellular expression of (R127W)HSPB1 and its co-localization with neurofilament light chain / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To observe the cellular expression of (R127W) HSPB1 and its influence on neurofilament light chain (NFL) self-assembly and co-localization with NFL.</p><p><b>METHODS</b>Eukaryotic expression vectors pEGFPN1-(wt) HSPB1 and pEGFPN1- (R127W) HSPB1 were constructed. Hela cells were transiently transfected with pEGFPN1-(wt) HSPB1 or pEGFPN1- (R127W) HSPB1 and observed under a confocal microscope. Hela cells were also transiently co-transfected with Pcl-NFL and pEGFPN1-(wt)HSPB1, or pCL-NFL and pEGFPN1-(R127W)HSPB1. The self-assembly of NFL was observed and the co-localization study of HSPB1/ (R127W)HSPB1 with NFL was carried out in these two cell models by immunofluorescence technique.</p><p><b>RESULTS</b>The aggregates formed by EGFP-(R127W)HSPB1 predominantly located around the nucleus, and EGFP-(wt)HSPB1 showed diffusion pattern in Hela cells. When co expressed with EGFP-(wt)HSPB1, NFL formed homogeneous structure in cytosol. When co-expressed with EGFP-(R127W)HSPB1, however, NFL had amorphous staining pattern predominantly consisting of NFL aggregates, and NFL co-localized with (R127W)HSPB1 in these aggregates.</p><p><b>CONCLUSION</b>The R127W mutant of HSPB1 may have reduced capacity to serve as a chaperone to prevent aggregate formation, and fail to correctly organize the neurofilament network. Dysfunction of the axon cytoskeleton and axon transport may be the primary mechanism of R127W mutation of HSPB1 in the pathogenesis of Charcot-Marie-Tooth disease.</p>

Effect of mutations in the 5'untranslated region pyrimidine-rich tract on the virulence variation of coxsackievirus B1 / 中国地方病学杂志

Objective To evaluate the infectivity and virulence variation caused by mutations in the 5' untranslated region(5'UTR)pyrimidine-rich tract of coxsackievirus B1(CVB1)genome.Methods Five pyrimidines in the 5'UTR pyrimidine-rich tract(nt563-nt573)of CVB1 genome were substituted with purines by site-directed mutagenesis.The mutant,CVB1/m563-573,was purified by plaque assay,and subjected to infectivity and virulence assessments by means of cytopathic effect(CPE),plaque forming,one-step growth curve,and 50% lethal dose(LD50)assays.Results Sequencing data revealed that the sequence of pyrimidine-rich tract in the 5'UTR of CVB1/m563-573 mutant was exactly identical to our design(C565A,U567C,U568A,U570A,and U572G).CPE assay showed that the infectivity of CVB1/m563-573 was weaker than that of its prototype CVB1/wt(A490=0.710±0.074,0.812±0.092)though no significant difference could be observed(t=-2.204,P>0.05).Plaque forming assay showed that the plaque quantities of CVB1/m563-573 were(6.40±1.52)×103,(11.60±2.19)×103 pfu/L and the plaque diameters of CVB1/m563-573 were(2.00±0.35),(2.47±0.41)mm at 46 and 58 hours pestinfection,respectively.The plaque quantities of CVB1/wt were(8.40±2.51)×103,(11.80±1.92)×103 pfu/L and the plaque diameters of CVB1/wt were(1.80±0.27),(2.85±0.44)mm,respectively.There was no significant difference between the plaque quantities and sizes of CVB1/m563-573 and CVB1/wt(t=8.000,0.985,10.000,9.000,all P>0.05).One-step growth curve demonstrated that the numbers(lg)of CVB1/m563-573 progenies at time-points of 3,5,7 h postinfection were 2.10±0.09,4.28±0.03,7.44±0 and that of CVB1/wt progenies were 2.80±0.02,4.77±0.02,8.55±0.01,respectively.The replication of CVB1/m563-573 was significantly slower than that of CVB1/wt at all three time-points(t=-13.151,-24.319,-47.714,all P<0.01).The LD50 of CVB1/m563-573(3.10×109 pfu/L)and CVB1/wt(1.26×107 pfu/L)indicated that the virulence of CVB1/m563-573 was significantly weakened compared to that of CVB1/wt.Conclusions The infectivity and virulence of CVB1 are weakened by substitution of pyrimidines with purines in the pyrimidine-rich tract of CVB1 5'UTR.Site-directed mutagenesis in the pyrimidine-rich tract may be a strategy for developing attenuated CVB vaccine.

Cardiac mast cells accumulation and degranulation contribute to collagen deposition after coronary microembolization / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate potential pathophysiological role of cardiac mast cells accumulation and degranulation on the collagen deposition after coronary microembolization (CME).</p><p><b>METHODS</b>CME was induced in miniswine by selective infusion of 15X10(4) microspheres (diameter, 45 mum) into the left anterior descending artery groups (CME group, n=8). Some CME-induced animals were pretreated with the MC stabilizer tranilast (50 mg/kg, twice daily), beginning 2 weeks before CME and thereafter throughout the experimental period (CME +tranilast group, n=8), while some animals received tranilast without CME (tranilast group, n=8). Eight sham-operated animals without CME served as controls. After 30 days, the total number of MC and degranulating MCs and collagen deposition was assessed by histological and electronic microscopy studies.</p><p><b>RESULTS</b>The numbers of total and degranulating MCs and collagen volume fraction (CVF) at day 30 in CME group were significantly higher than those in controls (P <0.01). Treatment with tranilast significantly reduced the numbers of total and degranulating MCs and CVF at day 30 (all P <0.01). There was a significant positive correlation of the CVF with the number of total MCs (r=0.91, P <0.001) and degranulating MCs (r=0.92, P <0.001) over the CME myocardium.</p><p><b>CONCLUSION</b>MCs accumulation and degranulating contribute to myocardial fibrosis collagen deposition.</p>