In this paper, Fourier transform infrared spectroscopy fingerprint analysis of Marsdenia tenacissima samples was used to develop a reliable method of tracing the geographical origins. Forty-eight samples from four provinces of China were analyzed by FTIR. We analyzed and characterized the fingerprints in both the full spectrum peaks and characteristic peaks, then the principal component analysis and the cluster analysis were carried out. The results of fingerprint analysis, correlation analysis, principal component analysis and cluster analysis can identify the geographic origins correctly, which verified and supplemented each other; the identification results and the actual location showed a high degree of consistency, namely the lower the space distance, the greater the similarity of different samples. These results revealed the obvious superiority and practical value in comparison to the more tedious and time-consuming wet chemistry method normally used. Using appropriate metrology methods can trace the geographical source correctly. The M. tenacissima materials from the region of Maguan should be considered as genuine medicinal materials taking into account the good quality.
China ; Cluster Analysis ; Drugs, Chinese Herbal ; analysis ; classification ; standards ; Geography ; Marsdenia ; chemistry ; classification ; Medicine, Chinese Traditional ; Principal Component Analysis ; Quality Control ; Reproducibility of Results ; Spectroscopy, Fourier Transform Infrared ; methods

Abstract: Objective　To understand the kdr (knockdown resistance, kdr) gene mutation of the voltage-gated sodium channel (VGSC) of Anopheles sinensis in Yunnan Province. Methods　From 2018 to 2019, mosquitoes were collected in Luoping County, Suijiang County, Tengchong City, Yingjiang County, Yuanjiang County and Mengla County in Yunnan Province. The collected mosquitoes were morphologically identified as Anopheles sinensis and genomic DNA was extracted by kits. The DNA templates were sequenced after PCR amplification and the sequencing results were identify as Anopheles sinensis by homology alignment in NCBI. After the ⅡS5 and ⅡS6 fragments of the sodium channels in Anopheles sinensis were amplified and sequenced, the sequencing results were multiple aligned by DNAMAN software, and the mutations were analyzed one by one with BioEdit software to determine the kdr allele types and genotypes, and the frequencies were calculated. Results This survey amplified 287 sequences, and the sequence maps showed that 1014 loci had three alleles, including wild type TTG/L (89.20%), mutant type TTT/F (9.76%) and TCG/S (1.04%). Five genotypes: homozygous wildtype L/L (85.02%), homozygous mutant F/F (6.27%) and S/S (0.35%), heterozygous mutant L/F (6.97%) and L/S (1.39%). The wild type allele TTG/L was the main allele in six sampling sites except Suijiang County. The frequency of wild type allele in Tengchong City was the highest (100.00%). That is, no mutation was detected, while the rest of counties occurred different degrees of mutation at 1014 loci. The frequency of mutant allele in Suijiang County was the highest, reaching 55.68%. Luoping County, Mengla County and Suijiang County had two mutant types. Yingjiang County and Yuanjiang County had one heterozygous mutant L/F. Conclusion　Wild type L1014 (TTG/L) is still dominant in most areas of Yunnan Province. The kdr mutation type is mainly L1014F, followed by L1014S, and the mutation frequency is lower than that in central provinces of China.

Objective To observe protective effects on rat serum cardiac enzymes and the antioxidant capacity of selenium and vitamin E.Methods According to body weight and 2 × 2 factorial design,eighty male Wistas rats were randomly divided into four groups:low selenium and low vitamin E group(feed containing 23.42％ of the low selenium yeast,excluding vitamin E),low selenium and adequate vitamin E group (feed containing 23.42％ of the low selenium yeast and vitamin E 160 mg/kg),adequate selenium and low vitamin E group(feed containing 46.84％ of the low selenium yeast and sodium seleni 0.25 mg/L in water,excluding vitamin E),adequate selenium and adequate vitamin E group(feed containing 46.84％ of the low selenium yeast,vitamin E 160 mg/kg and sodium selenite 0.25 mg/L in water),20 rats every group.Rats were feed with synthetic feed,and given intraperitoneal anesthesia after 26 weeks of feeding.Blood was collected to observe the impact of selenium and vitamin E on rat cardiac enzymes and myocardial antioxidant capacity and their interactions.Serum creatine kinase (CK) was measured using the continuous monitoring method,creatine kinase isozymes (CK-MB) and lactate dehydrogenase(LDH ) using the immune suppression method,the whole blood GSH-Px assay using the dithiobis nitrohenzoic acid(DTNB) method,serum superoxide dismutase(SOD) using the xanthine oxidase method,total antioxidant capacity (T-AOC) using the complex colorimetry method,the content of propylene glycol (MDA) using the thiobarbituric acid colorimetric method,and reactive oxygen species(ROS) using the colorimetric method.Results Group differences of serum CK,CK-MB,LDH,whole blood GSH-Px activity,serum T-AOC vitality,MDA and ROS content were statistically significant(F=9.797,17.041,48.399,3.744,224.900,49.384,5.045,all P＜ 0.05).Compared with the two low selenium groups and one adequate selenium group,the vitalities of CK,CK-MB,LDH and the contents of MDA[(1577.75 ± 451.87),(1239.15 ± 344.99),(884.25 ± 133.84)U/L,(5.688 ±1.169) × 103 nmol/L; (1474.21 ± 398.38),(1014.84 ± 215.40),(523.00 ± 98.05)U/L,(4.035 ± 0.487 ) × 103 nmol/L and (1180.10 ± 245.51),(948.75 ± 173.68),(676.70 ± 193.63)U/L,(3.406 ± 0.146) × 103 nmol/L]increased significantly in adequate selenium and adequate vitamin E group[( 1056.80 ± 250.98),(721.70 ±129.98),(404.65 ± 72.49)U/L,(3.010 ± 1.270) × 103 nmol/L,all P ＜ 0.05) ].The activity of GSH-Px was obviously increased in the two adequate selenium groups[ (96.611 ± 8.238) × 103,(103.024 ± 8.217) × 103 U/L,all P ＜ 0.05],compared with the two low selenium groups[ (60.356 ± 8.179) × 103,(63.117 ± 8.281) × 103 U/L].Selenium affected the activities of CK,CK-MB and LDH(F =27.09,31.58,29.66,all P＜ 0.01 ),and vitamin E affected the activities of CK-MB and LDH(F=18.9,11.2.all P＜ 0.01 ),but both selenium and vitamin E had no interactions on the activities of CK,CK-MB and LDH (F=0.02,0.001,2.22,all P＞0.05).Selenium affected the activity of GSH-Px and the content of MDA(F=6.74,95.68,all P＜ 0.05),vitamin E affected the activity of T-AOC,the contents of MDA and ROS(F=6.42,36.73,8.43,all P＜0.05),but selenium and vitamin E had interactions only on the content of MDA(F =13.82,P＜ 0.05).Conclusions Long-term selenium or vitamin E deficiency,can reduce the body's antioxidant capacity,leading to the occurrence of myocardial injury.Selenium and vitamin E can improve the body's oxidation capacity,playing a role in myocardial protection.

OBJECTIVETo establish a method for real-time recording the oxygen consumption of mice under normobaric hypoxia.

METHODSThe experimental apparatus was made up of animal container, filling water control system, electronic balance, hose, a computer with weight recording software, etc. The working principle was that the oxygen consumed by animal was replaced by water filling which was controlled by the pneumatic and hydraulic actuator. The water was weighted by an electronic balance and the weight signal was recorded into excel file at the same time. The accuracy and precision of the apparatus were detected by a 10 ml syringe. The oxygen consumption characteristics of 6 acute repetitive hypoxia mice and 6 normal mice were observed.

RESULTSThe P value for the paired t test was 1 and the CV value was 4%. The survival time and total oxygen consumption of acute repetitive hypoxia mice were both significantly increased compared to normal mice (P < 0.05), which were (58.8 +/- 6.8) min and (46.0 +/- 8.7) min respectively for the survival time and (85.1 +/- 8.5) ml and (73.6 +/- 5.4) ml respectively for total oxygen consumption.

CONCLUSIONThe hypoxia tolerance of the acute repetitive hypoxia mice can significantly improved by taking more oxygen in the animal cabin. The accuracy and precision of the apparatus are high and it can be used for the determination of oxygen consumption in hypoxia research.

Animals ; Hypoxia ; physiopathology ; Mice ; Monitoring, Physiologic ; instrumentation ; Oxygen Consumption ; physiology

OBJECTIVETo clone SABATH methyltransferase (rLjSABATHMT) gene in Lonicera japonica var. chinensis, and compare the gene expression and intron sequence of SABATH methyltransferase orthologous in L. japonica with L. japonica var. chinensis. It provide a basis for gene regulate the formation of L. japonica floral scents.

METHODThe cDNA and genome sequences of LjSABATHMT from L. japonica var. chinensis were cloned according to the gene fragments in cDNA library. The LjSABATHMT protein was characterized by bioinformatics analysis. SABATH family phylogenetic tree were built by MEGA 5.0. The transcripted level of SABATHMT orthologous were analyzed in different organs and different flower periods of L. japonica and L. japonica var. chinensis using RT-PCR analysis. Intron sequences of SABATHMT orthologous were also analyzied.

RESULTThe cDNA of LjSABATHMT was 1 251 bp, had a complete coding frame with 365 amino acids. The protein had the conservative SABATHMT domain, and phylogenetic tree showed that it may be a salicylic acid/benzoic acid methyltransferase. Higher expression of SABATH methyltransferase orthologous was found in flower. The intron sequence of L. japonica and L. japonica var. chinensis had rich polymorphism, and two SNP are unique genotype of L. japonica var. chinensis. The motif elements in two orthologous genes were significant differences.

CONCLUSIONThe intron difference of SABATH methyltransferase orthologous could be inducing to difference of gene expression between L. japonica and L. japonica var. chinensis. These results will provide important base on regulating active compounds of L. japonica.

Base Sequence ; Cloning, Molecular ; Computational Biology ; Evolution, Molecular ; Gene Expression Regulation, Plant ; Lonicera ; enzymology ; genetics ; Methyltransferases ; genetics ; Nucleotide Motifs ; Phylogeny ; Sequence Alignment ; Sequence Analysis

Humans ; Enhanced Recovery After Surgery ; Stomach Neoplasms/surgery* ; Length of Stay ; Laparoscopy ; Postoperative Complications ; Gastrectomy

OBJECTIVETo investigate the influence of chronic prostatitis on premature ejaculation (PE) and erectile dysfunction (ED) and the correlation of chronic prostatitis with PE and ED.

METHODSWe adopted NIH-CPSI, CISFPE and IIEF-5, interviewed 623 patients with chronic prostatitis, and recorded their scores on the above systems.

RESULTSThe incidence of PE was 39.0% and the rates of the mild, moderate and severe types were 26.2%, 12.0% and 0.8%, respectively. It developed mostly among the younger patients, accounting for 42.9% (103/240), 37.0% (95/257) and 35.7% (45/126) in the 18-30, 30-40 and 40-57 yr age groups, respectively. The incidence of ED was 16.9% and the rates of the mild, moderate and severe types were 14.9%, 1.0% and 1.0%, respectively. It occurred mostly among the older patients, accounting for 13.8% (33/240), 11.3% (29/257) and 34.1% (43/126) in the 18-30, 30-40 and 40-57 yr age groups, respectively. Analyses revealed no significant correlation among the scores on NIH-CPSI, CISFPE and IIEF-5.

CONCLUSIONThere is a high incidence of PE and ED among patients with chronic prostatitis, but without significant correlation between the severity of chronic prostatitis and that of PE and ED.

Adolescent ; Adult ; Chronic Disease ; Erectile Dysfunction ; epidemiology ; Humans ; Incidence ; Male ; Middle Aged ; Prostatitis ; epidemiology ; physiopathology ; Sexual Dysfunction, Physiological ; epidemiology ; Surveys and Questionnaires

OBJECTIVETo investigate the effects of 11, 12-epoxyeicosatrienoic acid (11, 12-EET) preconditioning and postconditioning on Ca(2+)-handling proteins in myocardial ischemia/reperfusion (IR) injury in rats and reveal the effects and mechanism of 11, 12-EET on cardioprotection. METHODS The IR injury model was built by stopping perfusion for 40 minutes followed by reperfusion for 30 minutes. The isolated Langendorff-perfused rat hearts were divided into 4 groups: control group, IR group, EET preconditioning (Pre-EET) group and EET postconditioning (Post-EET) group. The computer-based electrophysiological recorder system was used to measure the changes of the maximal rate of pressure increased in the contraction phase (+dp/dt(max)), the maximal rate of pressure decreased in the diastole phase (-dp/dt(max)), the left ventricular end diastolic pressure (LVEDP) and the difference of left ventricular pressure (delta LVP). The activity of Ca(2+)-ATPase in sarcoplasmic reticulum was measured with colorimetric method. Reverse transcription-polymerase chain reaction was used to assess the gene expression of C(a2+)-handling protein [sarcoplasic reticulum Ca(2+)-ATPase (SERCA), phospholamban (PLB), ryanodine receptor type 2 (RyR,), and 1, 4, 5-trisphosphate inositol receptor type 2 (IP3 R2) ] mRNAs level.

RESULTSCompared with IR group, the myocardial functions, the value of Ca(2+)-ATPase, and the expressions of IP3 R2 mRNA were significantly increased and the expression of PLB mRNA was significantly decreased in both Pre-EET group and Post-EET group (P < 0.05, P < 0.01). And the expression of SERCA mRNA was significantly increased in Pre-EET group (P < 0. 05). However, no significant differences were detected between Pre-EET and Post-EET groups. Moreover, the expression of RyR2 mRNA was not significantly different among all groups.

CONCLUSIONS11, 12-EET preconditioning and post-conditioning can protect myocardium from IR injury by elevating the activity of Ca(2+)-ATPase in sarcoplasmic reticulum, up-regulating the expression of IP3 R2 mRNA, and down-regulating the expression of PLB mRNA. Moreover, up-regulating the expression of SERCA mRNA maybe one of mechanisms of 11, 12-EET preconditioning on cardio protection against IR injury.

8,11,14-Eicosatrienoic Acid ; analogs & derivatives ; pharmacology ; Animals ; Calcium-Binding Proteins ; drug effects ; metabolism ; Inositol 1,4,5-Trisphosphate Receptors ; drug effects ; metabolism ; Ischemic Preconditioning, Myocardial ; methods ; Myocardial Reperfusion Injury ; metabolism ; prevention & control ; Rats ; Ryanodine Receptor Calcium Release Channel ; metabolism ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; drug effects ; metabolism

This study aimed to establish human IFN-gamma (hIFN-gamma) in vitro release assay and to apply it in diagnosis of human tuberculosis. Human IFN-gamma gene was cloned and expressed in Escherichia coli. The recombinant hIFN-gamma was purified and used as immunogen to immunize mice and rabbits respectively. Monoclonal and polyclonal antibodies were respectively developed and a sandwich ELISA was established. The heparized whole blood from 111 active tuberculosis patients and 292 clinical healthy controls were collected. The blood was stimulated with tuberculosis specific fused antigen ESAT-6/CFP-10 and the plasma was collected for IFN-gamma detection. The sensitivity for tuberculosis diagnosis was 95.5%, whereas the positive detection rate for the healthy controls was 16.7%. There was a significant difference between the patients and healthy controls (P<0.01) indicating that this assay had a high sensitivity and specificity, and thus could be promising in tuberculosis diagnosis.
Animals ; Antibodies, Monoclonal ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Interferon-gamma ; immunology ; secretion ; Mice ; Mice, Inbred BALB C ; Mycobacterium tuberculosis ; immunology ; Rabbits ; T-Lymphocytes ; immunology ; Tuberculosis ; diagnosis ; immunology

OBJECTIVETo determine the value of concurrent chemoradiotherapy in patients with locoregionally advanced nasopharyngeal carcinoma from the mainland of China.

METHODSData were extracted from randomized trials comparing chemotherapy plus radiotherapy with radiotherapy alone in locally advanced nasopharyngeal carcinoma. Actuarial rates of survival and distant metastases were calculated. The followed electronic databases were searched the Chinese Biomedicine database, Pubmed, Medline, Embase and Cochrane library; Data were extracted by tow reviewers and Review manager 4.1 software was applied for statistical analysis.

RESULTSEighteen trials with 1993 patients were include according to the including criterion. The 3-year overall survival rate of the chemoradiotherapy group and the radiotherapy group were 68.47% and 56.38% respectively, and the 5-year overall survival rate of the two groups above were 51.91% and 41.09% respectively, while the distant metastases rate of the chemoradiotherapy group and the radiotherapy group were 26.19% and 38.71% respectively. The result demonstrated that chemoradiotherapy increased overall survival by 12% at 3 years, and 11% at 5 years after treatment. After chemoradiotherapy, the rate of distant metastasis was reduce by 12%.

CONCLUSIONSIn patients with locoregionally advanced nasopharyngeal carcinoma, chemoradiotherapy significantly improves overall survival at 3 years, and 5 years compared with radiotherapy alone.

Antineoplastic Combined Chemotherapy Protocols ; China ; Combined Modality Therapy ; Humans ; Nasopharyngeal Neoplasms ; drug therapy ; radiotherapy ; Radiotherapy ; Randomized Controlled Trials as Topic ; Survival Rate