Regulation and influence of mitochondria on macrophages during hepatitis B virus infection / 中华肝脏病杂志

Hepatitis B virus (HBV) infection is an important public health concern, as approximately 3.5% of the world's population is currently chronically infected. Chronic HBV infection is the primary cause of cirrhosis, hepatocellular carcinoma, and deaths related to liver disease globally. Studies have found that in HBV infection, viruses can directly or indirectly regulate mitochondrial energy metabolism, oxidative stress, respiratory chain metabolites, and autophagy, thereby altering macrophage activation status, differentiation types, and related cytokine secretion type and quantity regulations. Therefore, mitochondria have become an important signal source for macrophages to participate in the body's immune system during HBV infection, providing a basis for mitochondria to be considered as a potential therapeutic target for chronic hepatitis B.

Protective effect of Luteolin on oxidative damage of retinal pigment epithelium cells by regulating the Nrf2/HO-1 pathway / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the protective effect and mechanism of luteolin on H2O2-induced oxidative damage of retinal pigment epithelium(RPE)cells. <p>METHODS:ARPE-19 cells were divided into the control group, H2O2 group, different doses of luteolin groups and Nrf2 inhibitor group, and the oxidative damage model of RPE was prepared by 100μmol/L H2O2, except for the control group. Cell activity was detected by Methyl thiazolyl tetrazolium(MTT)assay and proper experimental concentration of luteolin was determined. The cell morphology and activity was observed in each group. Cell apoptosis rate and reactive oxygen species(ROS)were detected by flow cytometry, malondialdehyde(MDA)and superoxide dismutase(SOD)were detected by kit method, and the expression of caspase-3, poly adeno-sine diphosphate ribose polymerase(PARP), B cell lymphoma-2(Bcl-2), nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxygenase-1(HO-1)proteins were detected by Western blot. <p>RESULTS: 100μmol/L luteolin has toxic effects on ARPE-19 cells, so 25μmol/L and 50μmol/L luteolin were selected for subsequent experiments. The cell activity, SOD activity and the protein expression levels of Bcl-2, Nrf2, HO-1 in 25μmol/L and 50μmol/L luteolin groups were significantly higher than the H2O2 group(<i>P</i><0.05). The apoptosis rate, ROS, MDA content and the protein expression levels of Caspase-3 and PARP in 25μmol/L and 50μmol/L luteolin groups were significantly lower than the H2O2 group(<i>P</i><0.05). The cell activity, SOD activity \〖(13.83±1.49)U/mL <i>vs</i>(22.69±1.83)U/mL\〗 and the protein expression levels of Bcl-2, Nrf2 and HO-1 protein expression in the Nrf2 inhibitor group were significantly lower than the 50μmol/L luteolin group(<i>P</i><0.05). The apoptosis rate, ROS, MDA content \〖(654.96±26.99)<i>vs</i>(446.52±29.42),(3.89±0.29)nmol/mL <i>vs</i>(2.06±0.19)nmol/mL\〗 and the protein expression levels of Caspase-3 and PARP in the Nrf2 inhibitor group were significantly higher than the 50μmol/L luteolin group(<i>P</i><0.05).<p>CONCLUSION: Luteolin can improve the oxidative damage of RPE cells induced by H2O2, and its mechanism may be related to the activation of the Nrf2/HO-1 pathway.

Analysis of gut microbiota diversity in the elderly based on high-throughput sequencing / 中华微生物学和免疫学杂志

Objective:To investigate the changes in gut microbiota diversity with age in elderly people using high-throughput sequencing.Methods:Ninety healthy volunteers were recruited. People who were <60 years old (middle-aged group) were set up as a baseline control group (Age A group), while those aged ≥60 years old were further divided into four groups (60-<70: Age B group, 70-<80: Age C group, 80-<90: Age D group, ≥90: Age E group). Fecal samples were collected to extract DNA. The second-generation sequencing technology was used to amplify and sequence the V3-V4 hypervariable region of 16S rDNA. Bioinformatics analysis was performed to compare the differences in gut microbiota and functional genes among groups.Results:At the phylum level, gut microbiota were composed mainly of Firmicute, Bacteroidetes, Proteobacteria and Actinobacteria in different groups. The proportion of Firmicute was the highest, accounting for over 60%, followed by that of Bacteroidetes. At the genus level, the abundance of Faecalibacterium genus decreased with age. The α diversity analysis showed that the gut microbiota in the elderly of different ages had higher abundance and uniformity, and there was no significant difference among groups. However, the β diversity analysis showed that in community structure there was difference between Age A and Age B groups, and similarity between Age B and Age C groups. Conclusions:The community structure of gut microbiota changed significantly between young and middle-aged people and the elderly over 60 years old. It tended to be relatively stable in people of 60-80 years old, but changed again when they were over 80 years old. Chronic inflammatory diseases, metabolic diseases and tumors in the elderly might be associated with the decrease in Faecalibacterium.

Early use of dexamethasone increases Nr4a1 in Kupffer cells ameliorating acute liver failure in mice in a glucocorticoid receptor-dependent manner / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

BACKGROUND AND OBJECTIVE@#Acute liver failure (ALF) is a type of disease with high mortality and rapid progression with no specific treatment methods currently available. Glucocorticoids exert beneficial clinical effects on therapy for ALF. However, the mechanism of this effect remains unclear and when to use glucocorticoids in patients with ALF is difficult to determine. The purpose of this study was to investigate the specific immunological mechanism of dexamethasone (Dex) on treatment of ALF induced by lipopolysaccharide (LPS)/D-galactosamine (D-GaIN) in mice.@*METHODS@#Male C57BL/6 mice were given LPS and D-GaIN by intraperitoneal injection to establish an animal model of ALF. Dex was administrated to these mice and its therapeutic effect was observed. Hematoxylin and eosin (H&E) staining was used to determine liver pathology. Multicolor flow cytometry, cytometric bead array (CBA) method, and next-generation sequencing were performed to detect changes of messenger RNA (mRNA) in immune cells, cytokines, and Kupffer cells, respectively.@*RESULTS@#A mouse model of ALF can be constructed successfully using LPS/D-GaIN, which causes a cytokine storm in early disease progression. Innate immune cells change markedly with progression of liver failure. Earlier use of Dex, at 0 h rather than 1 h, could significantly improve the progression of ALF induced by LPS/D-GaIN in mice. Numbers of innate immune cells, especially Kupffer cells and neutrophils, increased significantly in the Dex-treated group. In vivo experiments indicated that the therapeutic effect of Dex is exerted mainly via the glucocorticoid receptor (Gr). Sequencing of Kupffer cells revealed that Dex could increase mRNA transcription level of nuclear receptor subfamily 4 group A member 1 (Nr4a1), and that this effect disappeared after Gr inhibition.@*CONCLUSIONS@#In LPS/D-GaIN-induced ALF mice, early administration of Dex improved ALF by increasing the numbers of innate immune cells, especially Kupffer cells and neutrophils. Gr-dependent Nr4a1 upregulation in Kupffer cells may be an important ALF effect regulated by Dex in this process.

Gut microbiota composition and diversity in the elderly supported by institution-based care and home-based care / 中华微生物学和免疫学杂志

Objective To investigate the gut microbiota diversity between the elderly supported by institution-based care and home-based care. Methods Fresh stool samples were collected from 18 aged per-sons supported by institution-based care (G1 group), 20 aged persons with home-based care (G2 group) and 20 middle-aged and young adults (G3 group). The V3-V4 hypervariable region of 16S rDNA was ampli-fied and sequenced by next generation sequencing technology. Operational taxonomic units ( OTUs) were an-alyzed by QIIME analysis platform for species annotation, diversity analysis, and inter-group difference anal-ysis. Statistical analysis was performed using RStudio software. Results The top 6 microbiological taxa in the three groups were Firmicute, Bacteroidetes, Proteobacteria, Actinobacteria, Fusobacteria and Verru-comicrobia. The abundance of the Firmicute in the G1 and G2 groups showed significant differences [(61. 47±5. 58)％ vs (76. 55±3. 64)％, P<0. 05]. The G1 and G3 groups had a statistically significant difference in the abundance of the Proteobacteria [(9. 59±12. 68)％ vs (2. 15±2. 47)％, P<0. 05]. The abundance of both Bacteroidetes and Proteobacteria was higher in the G1 group than in the G2 group without significant difference between the two groups. No significant differences in diversity indices ( Shannon, Simpson and Chao1) were found between G1 and G2 groups (P>0. 05). Results of the NMDS analysis showed that the intra-group differences were greater than inter-group differences in G1 and G2 groups. Con-clusions No significant difference in the diversity of gut microbiota was detected between the elderly sup-ported by institution-based care and home-based care, but there were differences in the composition of the predominant gut microbiota.

Gut microbiota composition and diversity in the elderly supported by institution-based care and home-based care / 中华微生物学和免疫学杂志

Objective@#To investigate the gut microbiota diversity between the elderly supported by institution-based care and home-based care.@*Methods@#Fresh stool samples were collected from 18 aged persons supported by institution-based care (G1 group), 20 aged persons with home-based care (G2 group) and 20 middle-aged and young adults (G3 group). The V3-V4 hypervariable region of 16S rDNA was amplified and sequenced by next generation sequencing technology. Operational taxonomic units (OTUs) were analyzed by QIIME analysis platform for species annotation, diversity analysis, and inter-group difference analysis. Statistical analysis was performed using RStudio software.@*Results@#The top 6 microbiological taxa in the three groups were Firmicute, Bacteroidetes, Proteobacteria, Actinobacteria, Fusobacteria and Verrucomicrobia. The abundance of the Firmicute in the G1 and G2 groups showed significant differences [(61.47±5.58)% vs (76.55±3.64)%, P<0.05]. The G1 and G3 groups had a statistically significant difference in the abundance of the Proteobacteria [(9.59±12.68)% vs (2.15±2.47)%, P<0.05]. The abundance of both Bacteroidetes and Proteobacteria was higher in the G1 group than in the G2 group without significant difference between the two groups. No significant differences in diversity indices (Shannon, Simpson and Chao1) were found between G1 and G2 groups (P>0.05). Results of the NMDS analysis showed that the intra-group differences were greater than inter-group differences in G1 and G2 groups.@*Conclusions@#No significant difference in the diversity of gut microbiota was detected between the elderly supported by institution-based care and home-based care, but there were differences in the composition of the predominant gut microbiota.

Ethical considerations of cellular immunotherapy for cancer / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

With the rapid development of immunology, molecular biology, and associated technologies such as next-generation sequencing, cellular immunotherapy has recently become the fourth major cancer treatment. Immunotherapies based on T cells, natural killer cells, and dendritic cells play key roles in cancer immunotherapy. However, their application in clinical practice raises several ethical issues. Thus, studies should focus on proper adherence to basic ethical principles that can effectively guide and solve related clinical problems in the course of treatment, improve treatment effects, and protect the rights and interests of patients. In this review, we discuss cellular immunotherapy-related ethical issues and highlight the ethical practices and current status of cellular immunotherapy in China. These considerations may supplement existing ethical standards in cancer immunotherapy.

Evalation of Jingzhi Xiaoban Tablet in Improving Heart Function of Coronary Heart Disease Pa- tients by Doppler Tissue Imaging and Speckle Tracking Imaging Technology / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To evaluate the role of Jiangzhi Xiaoban Tablet (JXT) in improving heartfunction of coronary heart disease (CHD) patients by tissue Doppler imaging (TDI) and speckle trackingimaging (STI) technology.</p><p><b>METHODS</b>Recruited were 60 inpatients with confirmed CHD by coronary angiography at First Affiliated Hospital, Hunan University of Traditional Chinese Medicine from October 2013to November 2014. They were assigned to the treatment group (group A) and the control group (groupB) according to random digit table, 30 cases in each group. Patients in group A took JXT, 0.45 g/tablet,4 tablets each time, 3 times per day, while those in group B took Simvastatin Tablet, 20 mg/tablet, 1 tablet each time, once per evening. The therapeutic course for all was 8 weeks. The long axis view of theheart of 18 segments STI Peak strain LS and TDI peak systolic Sa parameters were performed in all patients before and after treatment.</p><p><b>RESULTS</b>Before treatment segments of STI strain LS and TDI longitudinal peak systolic peak Sa were not statistically different between the two groups (P > 0.05). Each segment of STI peak longitudinal strain LS and TDI peak systolic Sa in the two groups were higher after treatment than before treatment (P < 0.05). After treatment each segment of STI parameters of LS and eachTDI segment parameters of Sa were significantly lower in group B than in group A (P < 0.01).</p><p><b>CONCLUSION</b>JXT could improve heart function of CHD patients to different degrees, and its curative effect was betterthan that of routine Western medicine (Simvastatin Tablets) treatment.</p>

MicroRNA-21 protects cardiomyocytes from tumor necrosis factor-α induced apoptosis in vitro via modulating PTEN/AKT/FOXO3a pathway / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the effect of microRNA-21 on tumor necrosis factor (TNF)-α induced cardiomyocytes apoptosis and the association with PTEN/AKT/FOXO3a signaling pathway.</p><p><b>METHODS</b>Neonatal cardiomyocytes were isolated and cultured in vitro. Cardiomyocytes apoptosis was induced by TNF-α (10 ng/ml for 24 h) and examined by the cardiomyocytes apoptotic index. Eukaryotic expression vector for lenti-microRNA-21 was established and then transferred into the cardiomyocytes. MicroRNA-21 and PTEN mRNA were examined by qRT-PCR. Intracellular signal molecules, such as the expression of PTEN, phosphorylated PTEN, AKT, phosphorylated AKT (pAKTser473, pAKTThr308), FOXO3a, phosphorylated FOXO3a and FasL were detected by Western blot.</p><p><b>RESULTS</b>MicroRNA-21 reduced TNF-α induced cardiomyocytes apoptosis [(23.42 ± 1.98)% vs. (78.37 ± 2.03)%, P < 0.05]. TNF-α downregulated the expression of microRNA-21 and upregulated the mRNA and protein expressions of PTEN. Phosphorylation of PTEN, AKT and FOXO3a was enhanced in cardiomyocytes transfected with lenti-microRNA-21 (P < 0.05). TNF-α also significantly activated the phosphorylation of PTEN, AKT and FOXO3a (P < 0.05). Compared with cardiomyocytes treated with TNF-α (10 ng/ml), the phosphorylation of PTEN, AKT and FOXO3a as well as expression of pPTEN, pAKTser473, pFOXO3a and FasL were significantly suppressed in cardiomyocytes treated with lenti-microRNA-21 and TNF-α (P < 0.05). Total AKT and FOXO3a were similar among all groups (P > 0.05).</p><p><b>CONCLUSIONS</b>MicroRNA-21 could protect cardiomyocytes from TNF-α- induced apoptosis through PTEN/AKT/FOXO3a pathway, which might serve as a new therapy option for various cardiovascular diseases in the future.</p>

Expression of Six1 and Six4 in esophageal squamous cell carcinoma and their correlation with clinical prognosis / 中华病理学杂志

<p><b>OBJECTIVE</b>Six1 and Six4 are expressed in several tumors, and associated with tumor progress and poor prognosis. The aim of this study was to investigate the expression of Six1 and Six4 in esophageal squamous cell carcinoma (ESCC), and to evaluate their correlation with the clinicopathological factors and prognosis.</p><p><b>METHODS</b>Tissue microarray technology and immunohistochemical method (EnVision) were used to detect the expression of Six1 and Six4 in the tumor tissues and corresponding adjacent normal epithelium of esophagus from 292 ESCC patients.</p><p><b>RESULTS</b>Among the 292 ESCC patients, the positive rates of Six1 and Six4 protein expression in tumor tissues were 72.9% (213/292) and 56.2% (164/292), respectively, significantly higher than the expression rate of 33.2% (97/292) and 32.5% (95/292) in adjacent normal epithelium of esophagus (P < 0.05). Chi square test showed that the expression of Six1 protein was related to tumor size, depth of tumor invasion and patient survival status; higher Six4 protein expression level was related to poor differentiation and increased depth of invasion. Single factor Log-rank analysis revealed that gender, TNM stage, Six1 protein expression level were related to the overall survival of ESCC patients (P < 0.05), while the five-year survival rate was significantly higher in the Six1-negative group than the Six1-positive group [51.9% (41/79) vs. 43.7% (93/213)]. Multi-factor Cox proportional risk model analysis showed that TNM stage and positive expression of Six1 were independent prognostic factors for ESCC patients (P < 0.05).</p><p><b>CONCLUSIONS</b>Six1 and Six4 are highly expressed in ESCC. Their expression levels are closely related to the progress and prognosis of ESCC. Over-expression of Six1 is related to poor prognosis in ESCC patients. Thus, Six1 could be used as an important prognostic indicator for ESCC patients.</p>

Epidemiological survey of dentine hypersensitivity of 630 adults in rural of Sichuan province / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the prevalence and potential risk factors of dentine hypersensitivity of adults in rural of Sichuan province.</p><p><b>METHODS</b>All representative samples, including 630 adults living in rural of Sichuan Province, were selected by multi-stage, stratified and random sampling. The dentine hypersensitivity of all 630 cases was surveyed with questionnaire and oral clinical examination. SPSS 18.0 software was used for statistical analysis.</p><p><b>RESULTS</b>27.9% of all subjects were suffered from dentine hypersensitivity, sour was the most common stimulus of dentine hypersensitivity. The first premolar was the most common tooth with dentine hypersensitive, which occupied 27.4% of the total affected teeth. Female, acid regurgitation symptom, low frequency of toothbrush replacement (over 3 months), high tooth-brushing force and frequency of fresh fruits consumption (over 2 times per week) probably were high risk factors of dentine hypersensitivity.</p><p><b>CONCLUSION</b>The prevalence of dentine hypersensitivity occurs in rural of Sichuan province is high, thus for future the publicity and education on dentine hypersensitivity preventive should be strengthened.</p>

Detection of epidermal growth factor receptor mutations in non-small-cell lung carcinoma by direct sequencing and correlations with clinicopathological characteristics and sample types / 中华病理学杂志

<p><b>OBJECTIVE</b>To retrospectively analyze epidermal growth factor receptor (EGFR) gene mutation frequencies and distribution characteristics in Chinese patients with non-small-cell lung carcinoma (NSCLC) by direct gene sequencing.</p><p><b>METHODS</b>Clinical samples from 443 NSCLC patients were obtained for EGFR gene mutation analysis, including 299 surgical specimens, 59 core biopsies and 85 fine needle aspiration and pleural effusion cytology specimens. All samples were processed from paraffin embedded blocks and microdissection was performed to enrich tumor cells. PCR based direct gene sequencing was used to investigate tyrosine kinase domain coding region involving exon 18 through 21.</p><p><b>RESULTS</b>(1) Among 443 samples, 193 mutations were detected in 189 patients (42.7%) and 4 patients possessed two mutations involving two different exons in their tumor samples. The percentage of mutations involving exon 18 to 21 were 2.0% (4/193), 48.7% (94/193), 6.7% (13/193) and 42.5% (82/193) respectively. (2) There was no significant correlation of EGFR mutation with age, however, mutation rate (50.9%, 54/106) of exon 21 in patients over median age 57 was higher than that of the younger patients (32.2%, 28/87; P<0.01). (3) EGFR mutation rate was remarkably higher in female patients (53.5%, 107/200) than in male patients (33.7%, 82/243; P<0.01). (4) Mutation rate in adenocarcinomas (46.5%, 161/346) was much higher than in squamous cell carcinomas (13.3%, 4/30) and poorly differentiated carcinomas (24.1%, 7/29; P<0.01, P<0.05), while the adenosquamous carcinomas shared a mutation rate similar to that of adenocarcinoma (7/13, P>0.05). (5) In surgical samples, core biopsies and cytological samples, the EGFR mutation detection rates were 49.5% (148/299), 35.6% (21/59) and 23.5% (20/85) respectively. The fine needle aspiration and cytological samples showed much lower EGFR mutation detection rates (23.5%, 20/85) than that of surgical samples (49.5%, 148/299; P<0.01).</p><p><b>CONCLUSIONS</b>(1) Direct gene sequencing is a reliable and effective method for the detection of EGFR mutations in NSCLC, particularly for unknown EGFR mutations. (2) EGFR mutations are more frequent in female patients and patients with adenocarcinoma NSCLC, involving mainly exon 19 and 21. (3) The mutation distribution in exons of EGFR gene appears age-related. (4) Detection rate of EGFR mutation varies in different sample types. Small biopsy and fine needle aspiration specimens are valuable materials for analyzing EGFR mutation in NSCLC, although rare false negativity may occur using such samples.</p>

Role of liver X receptors on cardiomyocyte hypertrophy / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the expression of liver X receptors (LXR) in hypertrophic myocardium and the effect of LXR agonist T0901317 on angiotensin II (AngII) induced cardiomyocyte hypertrophy.</p><p><b>METHODS</b>Transverse aortic coarctation (TAC) or sham operation were performed in 2-month-old wide type mice (C57/B6). Two weeks later, the expression of LXR in myocardium was detected by quantitative real-time PCR analysis and Western blot analysis. The effect of LXR agonist T0901317 on AngII-induced hypertrophy in cultured neonatal rat cardiomyocytes was also assessed.</p><p><b>RESULTS</b>Quantitative real-time PCR analysis and Western blot analysis showed that LXRalpha but not LXRbeta expression was upregulated post TAC both at mRNA and protein levels (All P < 0.05). AngII induced increased [(3)H] leucine incorporation and cardiomyocyte hypertrophy were significantly reduced by T0901317 in a dose-dependent manner (P < 0.05). T0901317 also dose-dependently inhibited atrial natriuretic peptide (ANP) gene expression in cardiomyocytes (P < 0.05).</p><p><b>CONCLUSION</b>Our findings strongly suggest that LXR is a potent mediator of cardiomyocyte hypertrophy and LXR activation could attenuate AngII induced cardiomyocyte hypertrophy in vitro.</p>

Microsatellite instability of sporadic colorectal carcinomas and its clinicopathological significance / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the incidence of microsatellite instability (MSI) in sporadic colorectal carcinoma (CRC) using BAT-25 and BAT-26 loci, and its association with clinicopathological features.</p><p><b>METHODS</b>Microsatellite analysis was performed on tissue samples from 73 primary and 53 metastatic tumors collected at the Department of Pathology, Fudan University Cancer Hospital in 2002. Genomic DNA was extracted from paraffin-embedded tissues. Microsatellite alterations of BAT-25 and BAT-26 were detected using fluorescent PCR followed by fragment analysis on automatic DNA sequencer with GeneScan 3.1 software. A case of hereditary nonpolyposis colorectal cancer syndrome (HNPCC) with known high-frequency MSI (MSI-H) was included as positive control.</p><p><b>RESULTS</b>Eleven out of 73 samples from primary tumors (15.1%) were MSI-positive and significantly associated with patient age, tumor site, differentiation and prognosis (P < 0.05). There was no significant difference between the positive rate of MSI in tissue samples from primary and metastatic sites among the 53 metastatic tumors, being 17.0% and 13.2%, respectively, P > 0.05. Two cases with negative MSI at the primary site but positive at the metastatic sites were observed.</p><p><b>CONCLUSION</b>MSI is a frequent molecular event that may serve as a useful parameter for studying tumor biological behavior. MSI plays a partial role in the metastasis of sporadic CRC, but the role of mismatch repair genes and its exact mechanism remains to be determined. The classification of sporadic CRC according to MSI may be of importance both theoretically and practically.</p>

Two novel germline mutations of MLH1 in hereditary nonpolyposis colorectal cancer family / 中华病理学杂志

<p><b>OBJECTIVE</b>To explore germline mutations of MLH1 in hereditary nonpolyposis colorectal cancer (HNPCC), and to investigate the pathobiology of novel detectable mutations of MLH1.</p><p><b>METHOD</b>RNA was extracted from the peripheral blood of 12 patients from 12 different families fulfilling the Amsterdam II Criteria of HNPCC. Germline mutations of MLH1 were determined by RT-PCR with gene specific primers, heat-resistance reverse transcriptase and long-template PCR polymerase, followed by cDNA sequencing analysis. PCR-Genescan analysis was used to further investigate microsatellite instability with a panel of 5 microsatellite markers (BAT26, BAT25, D5S346, D2S123 and Mfd15), along with immunohistochemistry staining to detect the expression of MLH1 protein in the tumor tissues.</p><p><b>RESULTS</b>Four germline mutations were found in 4 patients, 2 of which were previously reported GTT-->GAT mutation at codon 384 of exon 12, and the other two were novel mutations: CGC-->TGC at codon 217 of exon 8 and CCG-->CTG at codon 581 of exon 16. Two tumors with the novel mutations had high frequency microsatellite instability showing more than 2 instable loci (RER + phenotype), and both tumors lost their MLH1 protein expression.</p><p><b>CONCLUSION</b>The two novel germline mutations of MLH1 identified in this study, i.e. CGC-->TGC at codon 217 of exon 8 and CCG-->CTG at codon 581 of exon 16, are very likely to have pathological significance.</p>

Effects of Xuezhikang on serum levels of high sensitive-C reactive protein, matrix metalloproteinase-9 and lipoprotein in patients with acute coronary syndrome / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effects of Xuezhikang (XZK) on serum levels of high sensitivity-C reactive protein (Hs-CRP), matrix metalloproteinase-9 (MMP-9) and lipoprotein in patients with acute coronary syndrome (ACS).</p><p><b>METHODS</b>Sixty-nine ACS patients were divided into the XZK group (40 cases) treated with conventional therapy and XZK and the control group (29 cases) with conventional therapy alone, another 30 healthy persons were assigned as the normal group. Before and after the treatment, the levels of Hs-CRP, MMP-9 and lipids were detected.</p><p><b>RESULTS</b>Compared with those in the normal subjects, Hs-CRP and MMP-9 levels in the ACS patients increased significantly (P <0.05), parallel with the extent of myocardial injury. After 2 weeks of XZK treatment, levels of Hs-CRP and MMP-9 of the XZK group decreased significantly (P <0.05), while lipids levels had no remarkable changes.</p><p><b>CONCLUSION</b>Hs-CRP and MMP-9 levels were closely correlated to the genesis and severity of ACS. Anti-inflammatory action of XZK plays an important role in early stage treatment of ACS patients.</p>

The analysis for mRNA mutation of MLH1, MSH2 genes and the gene diagnosis for hereditary nonpolyposis colorectal cancer / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify hereditary nonpolyposis colorectal cancer (HNPCC) families based on the germline mutations of MLH1 and MSH2 mRNA.</p><p><b>METHODS</b>RNA was extracted from the peripheral blood of the 14 members from 12 different families fulfilling Amsterdam Criteria II. The germline mutations of MLH1 and MSH2 mRNA were detected by cDNA sequencing analysis following reverse transcription-PCR(RT-PCR) with special primers, heat-resistance reverse transcriptase, and expand long template PCR. DNA was extracted from the peripheral blood of the 14 members, the corresponding exons, in which mutations were found using the above method, were amplified with Taq enzyme, sequencing analysis was followed.</p><p><b>RESULTS</b>Six germline mutations were detected and identified from the 6 different families based on mRNA, 4 of them to be in MLH1, the other 2 in MSH2. The MLH1 mutations distribute in the exon 8, 12, 16, and 19. The MSH2 mutations distribute in exons 1 and 2. The 6 mutations were identified from the corresponding exons respectively in genomic DNA sequencing analysis. The mutation types involve in 4 missense, 1 silent, and 1 non-coding area mutations. Five out of the 6 mutations have not been reported previously. Five out of the 6 mutations were pathological, involving in 5 different families. The five families were identified to HNPCC families.</p><p><b>CONCLUSION</b>HNPCC family can be identified with RNA-based sequencing of MLH1 and MSH2 from peripheral blood, which has the advantages of both cost, time saving and high sensitivity.</p>

Analysis of bcl-6 protein expression and gene rearrangement in diffuse large B-cell lymphoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate bcl-6 protein expression and gene rearrangement patterns in diffuse large B-cell lymphoma (DLBCL) and their clinicopathologic significance.</p><p><b>METHODS</b>Immunohistochemical studies for bcl-6 and CD10 proteins were performed on 51 cases of DLBCL paraffin-embedded tissues (including 22 nodal samples and 29 extranodal samples) and 10 cases of reactive lymphoid hyperplasia (RLH) paraffin-embedded tissues. Interphase fluorescence in-situ hybridization (FISH) with dual color breakapart probe was also used to identify rearrangement of bcl-6 gene in 32 cases of nodal DLBCL tissues (including 22 paraffin-embedded samples and 10 fresh samples) and 5 cases of RLH paraffin-embedded tissues.</p><p><b>RESULTS</b>(1) The rates of bcl-6 protein expression in nodal DLBCL, extranodal DLBCL and RLH were 72.7% (16/22), 75.9% (22/29) and 100.0% (10/10) respectively. The rates of CD10 expression were 40.9% (9/22), 41.4% (12/29) and 100.0% (10/10) respectively. All lymphoma samples which expressed CD10 also showed co-expression of bcl-6 protein. (2) The co-expression of bcl-6 and CD10 was observed in 40.9% (9/22) nodal DLBCL and 41.4% (12/29) extranodal DLBCL. Low clinical stage (stage I and II) was more frequently observed in cases with co-expression of bcl-6 and CD10 (P < 0.05). (3) The rates of bcl-6 gene rearrangement in nodal DLBCL was 28.1% (9/32), with 27.3% (6/22) in paraffin-embedded tissues and 30.0% (3/10) in fresh tissues. There was no statistically significant difference found between the two groups (P > 0.05). Bcl-6 gene rearrangement was not found in all the 5 cases of RLH, and there was a significant difference between RLH and DLBCL (P < 0.05).</p><p><b>CONCLUSIONS</b>The rate of bcl-6 protein expression is high in DLBCL cases, and the detection of bcl-6 and CD10 protein co-expression may help in the diagnosis and differential diagnosis of DLBCL. Those DLBCL cases with co-expression of bcl-6 and CD10 may also have a better prognostic implication. On the other hand, bcl-6 gene rearrangement can be identified by interphase FISH with dual color breakapart probe in both paraffin-embedded and fresh lymphoma tissues.</p>

Detection of FUS-CHOP fusion gene in paraffin-embedded tissues and its clinicopathologic significance for myxoid/round cell liposarcomas / 中华病理学杂志

<p><b>OBJECTIVE</b>To explore the feasibility of detecting FUS-CHOP fusion gene in formalin-fixed, paraffin-embedded tissue and its application in the diagnosis and differential diagnosis of myxoid/round cell liposarcomas (MRCLs).</p><p><b>METHODS</b>Forty-four formalin-fixed, paraffin-embedded MRCL samples and 60 control cases (atypical/well-differentiated liposarcoma, pleomorphic liposarcoma, low-grade myofibrosarcoma, etc.) retrieved from the archival files were studied. Nested reverse transcription-polymerase chain reaction (RT-PCR) technique was employed to detect the FUS-CHOP mRNA expression, followed by DNA sequencing confirmation of the PCR product. Housekeeping gene PGK was used to assess the quality of the mRNA templates.</p><p><b>RESULTS</b>PGK mRNA was detected in 93 of 104 tumor cases (89.4%), including 39 MRCLs cases (39/44, 88.6%) and 90% of the negative control cases. Type II FUS-CHOP fusion transcript was successfully detected in 20 out of 39 (51.3%) MRCL cases. Type I FUS-CHOP fusion transcript was not detected in any MRCLs in this study. All 60 negative control cases were negative for the FUS-CHOP fusion gene transcripts.</p><p><b>CONCLUSIONS</b>(1) Nested RT-PCR can be used to detect FUS-CHOP mRNA in formalin-fixed, paraffin-embedded tissues. (2) FUS-CHOP is considered a specific molecular and genetic hallmark for MRCLs. Nested RT-PCR is a sensitive and specific technique in detecting FUS-CHOP gene, and can be used in the diagnosis and differential diagnosis of MRCLs.</p>

Detection of cyclin D1 mRNA by reverse transcription-polymerase chain reaction in paraffin-embedded tissues and its diagnostic significance for mantle cell lymphoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of detecting cyclin D1 mRNA in paraffin-embedded tissues by reverse transcriptase polymerase chain reaction (RT-PCR) and competitive RT-PCR and its diagnostic and differential diagnostic significance for mantle cell lymphoma (MCL).</p><p><b>METHODS</b>Paraffin-embedded samples of 36 cases of MCL, 71 cases of other small B-cell lymphomas and 20 cases of lymphoid reactive hyperplasia as control group were retrieved from archival materials. Cyclin D1 protein and its mRNA was detected by EnVision and RT-PCR and competitive RT-PCR in all samples. House-keeping gene PGK was choosen as internal control.</p><p><b>RESULTS</b>(1) Cyclin D1 protein was expressed in 27 of the 38 MCL (71.1%). No cyclin D1 expression was found in the control group. (2) PGK was detected in 103 of the 116 cases (88.8%) and also detected in 34 of 36 MCL cases (94.7%). (3) cyclin D1 mRNA was detected in 34 nodal mantle cell lymphoma cases by RT-PCR in paraffin-embedded tissues. The positive rate of cyclin D1 mRNA was 94.4% in mantle cell lymphomas after exclusion of the 2 cases which were negative for both cyclin D1 mRNA and PGK. cyclin D1 mRNA was not detected in other nodal small B-cell lymphomas or lymphoid reactive hyperplasia, except 1 case of B-SLL. Sequencing analysis showed that sequences were identical to cyclin D1. (4) Cyclin D1 mRNA overexpression was detected in 27 cases of nodal mantle cell lymphoma by competitive RT-PCR in paraffin-embedded tissues. The positive rate of cyclin D1 mRNA overexpression was 75.0% in mantle cell lymphomas after exclusion of 2 cases which were negative for both cyclin D1 mRNA and PGK. cyclin D1 mRNA overexpression was not detected in other nodal small B-cell lymphomas or lymphoid reactive hyperplasia.</p><p><b>CONCLUSION</b>RT-PCR and competitive RT-PCR detection of cyclin D1 mRNA overexpression could be used for the diagnosis and differential diagnosis of mantle cell lymphoma in paraffin-embedded blocks.</p>