Analysis on Chemical Characteristics of Lycii Fructus in Qinghai Province Based on 1H-NMR Plant Metabolomics / 中国实验方剂学杂志

Objective:The chemical differences of Lycii Fructus samples from Qinghai, Ningxia, Gansu, Xinjiang and Inner Mongolia provinces were compared based on proton nuclear magnetic resonance (1H-NMR) plant metabolomics. Method:A total of 97 Lycii Fructus samples from five provinces were collected, including 61 samples in Qinghai, and extracted by 50% methanol for detecting. 1H-NMR spectra were obtained and compared by multivariate statistical analysis for investigating the chemical differences of samples from Qinghai and other production areas. And the content of Lycii Fructus polysaccharides in all samples was determined with the wavelength of 490 nm (calculated by anhydrous glucose). Result:A total of 32 chemical components were detected in the Lycii Fructus extract by 1H-NMR. The multivariate statistical analysis revealed that there was no significant difference among the samples from five provinces. The difference between Lycii Fructus from Qinghai and Ningxia, as well as the samples among the six regions of Qinghai province were relatively small. The similarity values of the majority of samples were >0.85. Univariate analysis showed that no significant difference was observed for the most metabolites in Lycii Fructus collected from five provinces, except for sucrose, glucose, proline and so on. There was no significant difference in the content of Lycii Fructus polysaccharides between Qinghai and other provinces. And the correlation coefficient between the content of Lycii Fructus polysaccharides and the small molecular compounds identified by 1H-NMR was -0.2-0.4. Conclusion:In this study, chemical characteristics of Lycii Fructus in Qinghai province are analyzed from the holistic view by 1H-NMR plant metabolomics, in combination of polysaccharide determination, and the results show that there is no significant difference between samples from Qinghai and other four provinces. The quality evaluation method based on 1H-NMR established in this study can provide scientific basis for improving quality control level and selecting planting areas of Lycii Fructus.

Clinical Phenotype and Gene Mutation Analysis of a Family with Early-Onset Familial Alzheimer′s Disease / 中山大学学报(医学科学版)

@#【Objective】A full exome sequencing of an early-onset family Alzheimer′s disease (EOFAD) was conduct? ed to identify the mutational sites which may cause diseases. The result of the current study may provide suggestion to genetic counseling and prenatal diagnosis.【Methods】Whole exome sequencing was performed on the family members and software PolyPhen-2 as well as SIFT was employed for hazard prediction (Prediction on functional effects of the missense mutation).【Results】The heterozygous mutation c.758A>G (p.Tyr253Cys) in exon 9 of TTC3 gene had been identified in proband whose mother had been proved with heterozygous mutation c.758A>G. According to the family separation and related bioinformatics analysis, the mutant gene was a possible pathogenic mutation. 【Conclusion】 A new mutation was found of c.758A>G in TTC3 gene within a Chinese EOFAD family and a new mutation to the spectrum of genetic mutation in EOFAD was expanded. The finding provides a significant groundwork for future exploration on the mechanisms underlying EOFAD.

Impacts of the regional cooperative chest pain center on the therapeutic time and the short term outcome of patients with ST segment elevated myocardial infarction / 中国介入心脏病学杂志

Objective To investigate the impact of the regional cooperative chest pain center (CPC) on therapeutic time and short term outcome after primary percutaneous coronary intervention (PCI) of patients with ST segment elevated myocardial infarction.Methods 372 patients with ST segment elevated myocardial infarction were enrolled in the study who had received were operated primary PCI 18 months before and after the regional cooperative CPC was set up.There were 156 patients in the green channel group before the setup of CPC and 216 patients in the CPC group.Total ischemia time,first medical contact (FMC) time,FML-to-balloon (FMC2B) time,door-to-balloon (D2B) time,hospital mortality rates,cardiac failure rates on the next day after PCI,length of CCU stays and hospital stays were compared between the two groups.Results Compared to the green channel group,total ischemia time[(281.0±102.7)min vs.(365.2±115.6)min,P＜0.05],FMC time [(174.3±97.5) min vs.(225.4±104.6) min,P＜0.05],FMC2B time [(106.7±61.2) min vs.(139.8±75.7) min,P＜0.05] and D2B time [(75.2±45.4) min vs.(102.4±53.7) min,P＜0.05] of the CPC group were significant shorter.The rates of reaching the standard of FMC2B time (70.83％ vs.34.62％,P＜0.001) and D2B time (75.93％ vs.40.38％,P＜0.001)were significantly higher in the CPC group.Cardiac failure rates on the next day after PCI was lower in the CPC group (14.35％ vs.23.72％,P=0.021),and CCU stays was shorter [(64.3±13.72)h vs.(92.6±15.65)h,P=0.043].Conclusions Establishment of a standardized regional cooperative CPC requires combination and consideration of the characteristics of local resources FMC2B time and D2B time of STEMI patients can be shorten by a standardized CPC lending to further shortening of total ischemia time and improvement in cardiac function.

Impact of Different Hospitals on Reperfusion Time in Acute ST-segment Elevation Myocardial Infarction Patients from Regional Cooperative Chest Pain Center / 中国循环杂志

Objective:To investigate the impact of different hospitals on reperfusion time in acute ST-segment elevation myocardial infarction (STEMI) patients from regional cooperative chest pain center (CPC).Methods:A total of 364 STEMI patients received percutaneous coronary intervention (PCI) at 18 months before and after CPC establishment were enrolled.Based on hospital levels,the patients were divided into 2 groups:Initial PCI hospital group,n=197 and Initial non-PCI hospital group,n=167.According to hospital visiting time,Initial PCI hospital group was further divided into 2 subgroups as Green channel subgroup,n=91 and CPC subgroup,n=106;Initial non-PCI hospital group was further divided into 2 subgroups as Routine referral subgroup,n=71 and CPC referral subgroup,n=96.Total ischemia time,from onset to first medical contact (S-FMC) time,from S-FMC to balloon dilatation (FMC2B) time,from hospital visit to balloon dilatation (D2B) time were compared among relevant patients;the impact factors for total ischemia time were studied by multivariate regression analysis.Results:Compared with Routine referral subgroup,the following parameters were shortened in CPC referral subgroup:total ischemia time 325 (236,1185) min vs 367 (214,1340) min,P＜0.05;FMC2B time 115 (82,227) min vs 149 (94,483) min,P＜0.05;D2B time 69 (35,195) min vs 105 (55,260) min,P＜0.05.Compared with CPC referral subgroup,the following parameters were further shortened in Initial PCI hospital group:total ischemia time 283 (168,873) min vs 325 (236,1185) min,P＜0.05;FMC2B time 78 (45,265) min vs 115 (82,227) min,P＜0.05.Multivariate linear regression analysis presented that high school or above education (β=-0.117,P=0.047),arrived PCI hospital within 60 min of onset (β=-0.243,P=0.000)and using initial PCI hospital (β=-0.175,P=0.000) were the independent impact factors for total ischemia time in STEMI patients.Conclusion:Regional cooperative CPC may shorten FMC2B time by patients' referral;visiting PCI hospital within 60 min of onset was the best way to reduce total isehemia time in STEMI patients.

Impact of Different Hospitals on Reperfusion Time in Acute ST-segment Elevation Myocardial Infarction Patients from Regional Cooperative Chest Pain Center / 中国循环杂志

Objective:To investigate the impact of different hospitals on reperfusion time in acute ST-segment elevation myocardial infarction (STEMI) patients from regional cooperative chest pain center (CPC).Methods:A total of 364 STEMI patients received percutaneous coronary intervention (PCI) at 18 months before and after CPC establishment were enrolled.Based on hospital levels,the patients were divided into 2 groups:Initial PCI hospital group,n=197 and Initial non-PCI hospital group,n=167.According to hospital visiting time,Initial PCI hospital group was further divided into 2 subgroups as Green channel subgroup,n=91 and CPC subgroup,n=106;Initial non-PCI hospital group was further divided into 2 subgroups as Routine referral subgroup,n=71 and CPC referral subgroup,n=96.Total ischemia time,from onset to first medical contact (S-FMC) time,from S-FMC to balloon dilatation (FMC2B) time,from hospital visit to balloon dilatation (D2B) time were compared among relevant patients;the impact factors for total ischemia time were studied by multivariate regression analysis.Results:Compared with Routine referral subgroup,the following parameters were shortened in CPC referral subgroup:total ischemia time 325 (236,1185) min vs 367 (214,1340) min,P＜0.05;FMC2B time 115 (82,227) min vs 149 (94,483) min,P＜0.05;D2B time 69 (35,195) min vs 105 (55,260) min,P＜0.05.Compared with CPC referral subgroup,the following parameters were further shortened in Initial PCI hospital group:total ischemia time 283 (168,873) min vs 325 (236,1185) min,P＜0.05;FMC2B time 78 (45,265) min vs 115 (82,227) min,P＜0.05.Multivariate linear regression analysis presented that high school or above education (β=-0.117,P=0.047),arrived PCI hospital within 60 min of onset (β=-0.243,P=0.000)and using initial PCI hospital (β=-0.175,P=0.000) were the independent impact factors for total ischemia time in STEMI patients.Conclusion:Regional cooperative CPC may shorten FMC2B time by patients' referral;visiting PCI hospital within 60 min of onset was the best way to reduce total isehemia time in STEMI patients.

Berberine enhances antidiabetic effects and attenuates untoward effects of canagliflozin in streptozotocin-induced diabetic mice / 中国天然药物

The present study aimed at determining whether berberine can enhance the antidiabetic effects and alleviate the adverse effects of canagliflozin in diabetes mellitus. Streptozotocin-induced diabetic mice were introduced, and the combined effects of berberine and canagliflozin on glucose metabolism and kidney functions were investigated. Our results showed that berberine combined with canagliflozin (BC) increased reduction of fasting and postprandial blood glucose, diet, and water intake compared with berberine or canagliflozin alone. Interestingly, BC showed greater decrease in blood urea nitrogen and creatinine levels and lower total urine glucose excretion than canagliflozin alone. In addition, BC showed increased phosphorylated 5' AMP-activated protein kinase (pAMPK) expression and decreased tumor necrosis factor alpha (TNFα) levels in kidneys, compared with berberine or canagliflozin alone. These results indicated that BC was a stronger antidiabetic than berberine or canagliflozin alone with less negative side effects on the kidneys in the diabetic mice. The antidiabetic effect was likely to be mediated by synergically promoting the expression of pAMPK and reducing the expression of TNFα in kidneys. The present study represented the first report that canagliflozin combined with berberine was a promising treatment for diabetes mellitus. The exact underlying mechanisms of action should be investigated in future studies.

Molecular cloning and characterization of Pseudorabies virus EP0 gene.

The pseudorabies virus (PRV) early protein EP0 is a homologue of the herpes simplex virus 1 (HSV-1) immediate-early protein ICP0, which is a multifunctional protein and important for HSV-1 infection. However, the exact function of EP0 is not clear. In this study, using polymerase chain reaction, a 1,104 base-pair sequence of the EP0 gene was amplified from the PRV Becker strain genome and identification of the EP0 gene was confirmed by further cloning and sequencing. Bioinformatics analysis indicated that the PRV EP0 gene encoded a putative polypeptide with 367 amino acids. The encoded protein, designated as EP0 contained a conserved RING-finger superfamily domain and was found to be closely related with the herpes virus RING-finger superfamily and was highly conserved among the counterparts encoded by RING-finger genes. Multiple nucleic acid sequence and amino-acid sequence alignments suggested that PRV EP0 showed a relatively higher similarity with EP0-like proteins of genus Varicellovirus than with those of other genera of Alphaherpesvirinae. In addition, phylogenetic analysis showed that PRV EP0 had a close evolutionary relationship with members of genus Varicellovirus, especially bovine herpesvirus 1 (BoHV-1) and BoHV-5. Antigen prediction indicated that several potential B-cell epitopes were located in EP0. Also, subcellular localization analysis demonstrated that EP0 was predominantly localized in the nucleus, suggesting that it might function as a nuclear-targeted protein.

Mesenchymal stem cells transplanted in mdx mice differentiate into myocytes and express dystrophin/utrophin / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the differentiation of rat bone marrow mesenchymal stem cells (MSCs) into myocytes and their expression of dystrophin/utrophin after transplantation in mdx mice.</p><p><b>METHODS</b>BrdU-labeled fifth-passage rat MSCs were transplanted in mdx mice with previous total body gamma irradiation (7 Gy). At 4, 8, 12 and 16 weeks after the transplantation, the mice were sacrificed to detect dystrophin/BrdU and utrophin expressions in the gastrocnemius muscle using immunofluorescence assay, RT-PCR and Western blotting. Five normal C57 BL/6 mice and 5 mdx mice served as the positive and negative controls, respectively.</p><p><b>RESULTS</b>Four weeks after MSC transplantation, less than 1% of the muscle fibers of the mdx mice expressed dystrophin, which increased to 15% at 16 weeks. Donor-derived nuclei were detected in both single and clusters of dystrophin-positive fibers. Some BrdU-positive nuclei were centrally located, and some peripherally within myofibers. Utrophin expression decreased over time after transplantation.</p><p><b>CONCLUSION</b>The myofibers of mdx mice with MSC transplantation express dystrophin, which is derived partially from the transplanted MSCs. Dystrophin expression from the transplanted MSCs partially inhibits the upregulation of utrophin in mdx mouse muscle, showing a complementary relation between them.</p>

Induction of Apoptosis of Human Colon Cancer Cells by siRNA Recombinant Expression Vector Targeting Survivin Gene / 华中科技大学学报（医学）（英德文版）

This study constructed siRNA recombinant expression vector targeting survivin gene and observe the apoptosis induction effect of it in human colon cancer cells,siRNA recombinant expres-sion vector targeting survivin gene was constructed and transfected into human colon cancer cells.The effect of siRNA recombinant expression vector was detected by RT-PCR,Western blot,MTT re-duction assay and flow cytometry.It was confirmed by restriction endonuclease and sequence analy-sis that siRNA recombinant expression vector targeting survivin gene was constructed successfully.Inhibition rate of survivin siRNA at mRNA and protein levels was 36.33% and 44.65% respectively.Growth of cancer cells was inhibited and the apoptosis rate was (17.24±2.13)%.The siRNA recom-binant expression vector targeting survivin gene has been constructed successfully.It not only can in-hibit the expression of survivin gene,but also can induce apoptosis in human colon cancer cells re-markably.

Dynamic distribution of implanted human bone marrow mesenchymal stem cells in mdx mice / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the dynamic distribution of human bone marrow mesenchymal stem cells (hBM-MSCs) in mdx mice.</p><p><b>METHODS</b>Twenty-four 8-10-week-old immunocompromised mdx mice were transplanted with 1 x 10(7) passage 5 hBM-MSCs labeled with bromodeoxyuridine (BrdU) by means of injection into the tail vein. The mice were euthanized 48 hours and 2, 4, 8, 12, 16, 20, and 24 weeks after transplantation. BrdU-positive cells in tissue and organs of the mice were detected by immunofluorescence analysis. Skeletal muscle was stained for anti-human nuclei mouse monoclonal antibody (anti-Hu) and analyzed for human dystrophin (Dys) expression by immunohistochemistry and reverse transcription-polymerase chain reaction.</p><p><b>RESULTS</b>After transplantation, BrdU-positive cells were found in most organs (especially in bone marrow, liver, and lung) within 4 weeks, and these cells in liver and lung decreased gradually after 4 weeks. At 48 hours after transplantation, BrdU-positive cells were found in bone marrow, which reached a peak level after 2 weeks and were still detectable after 16 weeks. BrdU-positive cells in skeletal muscle increased gradually over time of transplantation. A small number of anti-Hu positive cells were detected in skeletal muscle 2 weeks after transplantation. A small number of Dys positive cell were seldom found at 4 weeks and small Dys mRNA expression detected 4 weeks after transplantation. The proportion of anti-Hu in parallel with Dys positive cells and Dys mRNA in skeletal muscle of mdx mice increased gradually over time of transplantation.</p><p><b>CONCLUSION</b>After being transplanted into mdx mice, hBM-MSCs are mainly distributed in bone marrow, liver, and lung during the early time (2-4 weeks) , and then in bone marrow and skeletal muscle (after 4 weeks).</p>

Association between angiotensin-converting enzyme and polymorphisms of N5, N10-methylenetetrahydrofolic acid reductase gene in patients with ischemic stroke / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the association between angiotensin-converting enzyme (ACE) and the polymorphisms of N5, N10-methylenetetrahydrofolic acid reductase (MTHFR) gene in patients with ischemic stroke (IS).</p><p><b>METHODS</b>Totally 454 patients with IS (IS group) and 334 controls (control group) were recruited in our study. Their I/D polymorphisms of ACE gene and C677T polymorphisms of MTHFR gene were detected by PCR and denaturing high performance liquid chromatography.</p><p><b>RESULTS</b>The frequencies of DD, ID, II and CC, CT, TT genotype in IS group were 22.5%, 43.4%, 34.1%, and 51.8%, 40.5%, 7.7%, respectively, and were 17.4%, 45.5%, 37.1% and 56.9%, 38.3%, 4.8% in the control group, respectively. DD genotype was associated with large-artery atherosclerosis (LAA), and TT genotype and T allele were associated with LAA and cardioembolism. Synergistic effects were found between TT and DD/ID DD genotypes in the pathogenesis of ischemic stroke.</p><p><b>CONCLUSION</b>DD, TT genotype and T allele are risk factors of IS, and ACE gene and MTHFR gene have synergistic effects in the pathogenesis of IS.</p>

Detecting the polymorphism of methylenetetrahydrofolate reductase gene by denaturing high performance liquid chromatography / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish a method for detecting the polymorphism of methylenetetrahydrofolate reductase gene (MTHFR).</p><p><b>METHODS</b>The MTHFR was amplified, and the amplified products were detected by denaturing high performance liquid chromatography (DHPLC), and the amplified MTHFR was confirmed by sequencing and restriction enzyme digesting.</p><p><b>RESULTS</b>A total of 334 individuals of Han people in southern China were recruited in our study, and their polymorphisms of MTHFR were detected. The accurate rate of the DHPLC method, that was very sensitive with 100% detection rate available, was over 99%. The frequencies of CC, CT and TT genotypes were 56.9%, 38.3% and 4.8% individually, and the frequencies of T and C alleles were 23.95% and 76.05% individually.</p><p><b>CONCLUSION</b>The DHPLC method can detect polymorphism of MTHFR rapidly, effectively and economically. And there is the existence of different MTHFR polymorphisms in area and race.</p>

Malignant hypertrophic cardiomyopathy caused by the Arg723Gly mutation in beta-myosin heavy chain gene in a Chinese pedigree / 中华心血管病杂志

<p><b>OBJECTIVE</b>Hypertrophic cardiomyopathy (HCM) is a genetically and phenotypically heterogeneous disease and an Arg723Gly mutation in beta-myosin heavy chain (beta-MHC) gene was found in 3 Spanish families with malignant HCM. We detected this gene mutation in 5 Chinese pedigrees with hypertensive cardiomyopathy.</p><p><b>METHODS</b>Five Chinese pedigrees with HCM and 80 age-matched normal control subjects were chosen for the study. The exons in the functional regions of the beta-MHC gene were amplified with PCR and the products were sequenced, genotype and phenotype analyzed.</p><p><b>RESULTS</b>Arg723Gly mutation was identified in exon 20 in one pedigree. In this pedigree, 13 out of 25 family members were diagnosed as HCM, 5 died of heart failure, all HCM patients in this pedigree had Arg723Gly mutation and 3 of them had NYHA III and 2 of them were diagnosed as HCM before the age of 20.</p><p><b>CONCLUSIONS</b>Arg723Gly mutation was also one of the main disease-causing genes in Chinese familial HCM. The mutation of Arg723Gly is a malignant phenotype as shown by early progressive heart failure development and poor prognosis in this pedigree with Arg723Gly mutation.</p>

Mutation of Arg723Gly in beta-myosin heavy chain gene in five Chinese families with hypertrophic cardiomyopathy / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Hypertrophic cardiomyopathy (HCM) is a form of cardiomyopathy with an autosomal dominant inherited disease, which is caused by mutations in at least one of the sarcomeric protein genes. Mutations in the beta-myosin heavy chain (beta-MHC) are the most common cause of HCM. This study was to reveal the disease-causing gene mutations in Chinese population with HCM, and to analyze the correlation between the genotype and phenotype.</p><p><b>METHODS</b>The exons 3 to 26 of MYH7 were amplified by PCR, and the PCR products were sequenced in five non-kin HCM patients. A 17-year-old patient was detected to be an Arg723Gly mutation carrier. Then his family was gene-screened, and the correlation between genotype and phenotype was analyzed.</p><p><b>RESULTS</b>The mutation of Arg723Gly in a Chinese family with HCM was detected for the first time. With a C-G transversion in nucleotide 13,619 of the MYH7 gene, located at the essential light chain interacting region in S1, the replacement of arginine by glycine took place at amino acid residue 723. A two-dimensional echocardiogram showed moderate asymmetrical septal hypertrophy with left atria enlargement. There was no obstruction in the left ventricular outflow tract. In his family, a total of 13 individuals were diagnosed HCM and 5 of them were dead of congestive heart failure at a mean age of 66-year-old. Eight living members were all detected to carry the mutation, in which 3 developed progressive heart failure. Moreover, the heart function of the people evidently deteriorates when their age are older than 50. The mutation and the disease show co-separated.</p><p><b>CONCLUSION</b>The Arg723Gly mutation is a malignant type. In Chinese the mutation has the similar characters to the former report but has low degree malignant.</p>

Expression of ITF Gene in Lacococcus lactis / 中国生物工程杂志

To develop an oral drug,ITF gene encoding ITF proein,was expressed in a live delivery vehicle lactococcus lactis.First,the ITF gene was cloned into the prokaryotic expressive vector pNICE:sec.Second,the recombinant vector pNICE:sec-ITF was transformated into Lactococcus lactis strain NZ9000 to express ITF protein.Then the recombinant ITF was induced to express and was identified by SDS-PAGE and Western blot.Rabbits are divided into blank control group,preparation group and therapeutic group which are respectively administrated wih PBS and pNICE:sec-ITF Lactococcus lactis.By grades of ulcer test whether administrated pNICE:sec-ITF Lactococcus lactis protects against HCl-induced gastric injury in rabbits.The results were described as follows.The ITF was amplified and cloned in the vector pNICE:sec successfully.The fusion protein(5.9kDa) was expressed in L.lactis by the induction of the nisin.The quantity of expression accounted for 5% of the total bacterial protein.Western bolt analysis confirmed that fusion protein could be recognized specially by Monoclonal Anti-human TFF3 Antibody.Preparation groups and therapeutic groups do good than control group.Prove that administrated pNICE:sec-ITF Lactococcus lactis is biologically active in an HCl-induced rabbit gastric mucosal injury model.

Experimental and clinical research of "Keyouling" on treatment of condyloma acuminata / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the effects of traditional Chinese medicine "Keyouling" in treatment of condyloma acuminata (CA).</p><p><b>METHODS</b>The effects of "Keyouling" at different concentrations on growth and proliferation of rabbits' prepuce epithelial cells and CA warts tissue were assessed by means of cellular primary culture and subculture, and the effects of "Keyouling" at different concentrations on the expression level of HPV6.11 DNA and expression level of HPV6.11 DNA of different area of CA patient's penis were quantitatively detected by means of FQ-PCR.</p><p><b>RESULTS</b>"Keyouling" had no effect on growth and proliferation of rabbits' prepuce cuticle cells, but it had significant inhibition on CA warts tissue cells and expression of HPV6.11 DNA. Concentrations of "Keyouling" assumed negative correlated with cell proliferation rate of CA warts tissue and expression level of HPV6.11 DNA, and also had significant inhibition on expression of HPV6.11 DNA of negative cutaneous lesion area and cutaneous lesion area of CA patients.</p><p><b>CONCLUSION</b>"Keyouling" had significant anti HPV6.11 effect, could significantly restrain growth and proliferation of CA warts tissue cells, while it had no damaging effect on normal organic cuticle cells. It was suggested that "Keyouling" was safe in clinical use and had good therapeutic effect on CA and subclinical infection.</p>