Medicinal plant germplasm resources are the foundation of the modern development of traditional Chinese medicine. In-depth study of medicinal plant germplasm resources is a prerequisite for cultivating fine varieties and ensuring the output and standard quality of traditional Chinese medicine（TCM）. Traditional identification methods start with appearance and are greatly affected by natural environment and human factors，with a low efficiency and accuracy of identification are generally low molecularin general. Due to such advantages as easy operation，high sensitivity，accurate results， molecular biology technology has been widely used in the related research of relevant studies for medicinal plant germplasm resources due to its advantages of easy operation，high sensitivity，accurate results，etc. It mainly involving the distinction between wild and cultivated products，researchstudy on substitutes of TCM，identification of Chinese patent medicine，good variety marker breeding，genetic diversity researchstudy，genetic map establishment and omics research，etcstudy. Among them，omics researchstudy is divided into genomics，transcriptomics，metabolomics，and proteomics due toby different analysis purposes. Genomics is divided into three sub-fields namely structural genomics，functional genomics， and comparative genomics. Eukaryotes Because eukaryotes have nuclei and organelles，so omics researchstudy also includes chloroplast genomics，mitochondrial genomics，nuclear genomics，and plastid genomics. Among them，the chloroplast genome has a simple structure，small molecular weight，and good conservation，while the mitochondrial genome has a strong variability and complex structure，the nuclear genome data isfeatures complex， data and the nucleus contains no ribosomes in nucleus，resulting in spatiotemporal differences in the translation process，even if repeated repeatedly test， the result of and the test is alsoresults remained uncertain， even after repeated tests. The molecular biology technology and omics researchstudy involved in theby current medicinal plant researchstudy still hashave shortcomings，and there iswith a large room for development，which needs and need further improvement and supplementation. This articlepaper successively introduces the characteristics and applications of cytology，molecular markers，and omics researchstudy techniques in the identification of medicinal germplasm resources，providingin order to provide a reference for subsequent identification，development and utilization of medicinal plant germplasm resources.

Objective:To develop the specific molecular markers of Codonopsis plants and better identify their germplasm resources considering the significant difference in active ingredients of Codonopsis Radix from various origins and producing areas. Method:Such bioinformatics software as Primer 5.0， NTSYS-pc 2.10e， and PopGene 32 were used for searching the simple sequence repeat （SSR） markers of C. minima chloroplast genome， C. tsinlingensis chloroplast， and C. lanceolata mitochondrial sequences， and 120 pairs of SSR primers were designed by Primer 5.0. Then 16 pairs of cpSSR primers and 10 pairs of mtSSR primers with good screening effect and high polymorphism were selected for analyzing the interspecific versatility of 20 samples. Result:The results showed that 66 cpSSR primer sites and 26 mtSSR sites were identified from the genome sequences， with 86.20% of single nucleotide， 6.9% of dinucleotide， and 6.9% of trinucleotide for C. minima chloroplast， 83.78% of single nucleotide，13.51% of dinucleotide， and 2.71% of trinucleotide for C. tsinlingensis chloroplast， and 46.15% of single nucleotide and 53.85% of dinucleotide for C. lanceolata mitochondria. As demonstrated by polymerase chain reaction （PCR） identification results， the developed 26 pairs of SSR primers had good applicability in the genus Codonopsis. The analysis by NTSYS-pc 2.10e revealed that the genetic similarity coefficients of 20 samples were within the range of 0.38-1.00， and they were divided into two subgroups at a threshold of 0.69. Four pairs of polymorphic primers were screened out in the diversity analysis of 20 samples using PopGene 32. The number of observed alleles （Na） was 12， and the effective number of alleles （Ne） ranged from 1.362 9 to 2.605 9. The percentage of polymorphic loci （PPL） at each site was 100%， and the average values of genetic parameters Ho， He， and I at each site were 0.555 8， 0.444 2， and 0.753 2， respectively， indicating high polymorphism at each site. The screened four pairs of primers were utilized for DNA fingerprinting of the 20 samples， and it was found that the DNA fingerprints enabled the identification of these 20 samples. Conclusion:This study has provided a molecular basis for the study of the genetic relationship between plants in species Codonopsis and the intraspecific genetic differentiation.

Objective:To observe the effect of extracorporeal shock wave therapy on tendon adhesion in late period after hand tendon repair. Methods:From July, 2017 to December, 2018, 40 patients with tendon adhesion after hand tendon repair more than three months were collected. They were randomly divided into control group (n = 20) and experimental group (n = 20). Two groups received routine therapy, and the experimental group added extracorporeal shock wave therapy. Before and two months after treatment, the total active movement (TAM) of the fingers and the grip strength were messured. Results:There was no significant difference in TAM of the fingers and the grip strength before treatment (P > 0.05). After treatment, TAM of the fingers and the grip strength significantly increased (|t| > 10.284,P < 0.001), and were higher in the experimental group than in the control group (t > 0.386,P < 0.001). Conclusion:Extracorporeal shock wave therapy could facilitate to improve the tendon slippage and hand function in patients with tendon adhesion after hand tendon repair.

BackgroundLipoxin A4 (LXA4) can alleviate lipopolysaccharide (LPS)-induced acute lung injury (ALI) and acute respiratory distress syndrome through promoting epithelial sodium channel (ENaC) expression in lung epithelial cells. However, how LXA4 promote ENaC expression is still largely elusive. The present study aimed to explore genes and signaling pathway involved in regulating ENaC expression induced by LXA4.

MethodsA549 cells were incubated with LPS and LXA4, or in combination, and analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) of ENaC-α/γ. Candidate genes affected by LXA4 were explored by transcriptome sequencing of A549 cells. The critical candidate gene was validated by qRT-PCR and Western blot analysis of A549 cells treated with LPS and LXA4 at different concentrations and time intervals. LXA4 receptor (ALX) inhibitor BOC-2 was used to test induction of candidate gene by LXA4. Candidate gene siRNA was adopted to analyze its influence on A549 viability and ENaC-α expression. Phosphoinositide 3-kinase (PI3K) inhibitor LY294002 was utilized to probe whether the PI3K signaling pathway was involved in LXA4 induction of candidate gene expression.

ResultsThe A549 cell models of ALI were constructed and subjected to transcriptome sequencing. Among candidate genes, N-myc downstream-regulated gene-1 (NDRG1) was validated by real-time-PCR and Western blot. NDRG1 mRNA was elevated in a dose-dependent manner of LXA4, whereas BOC-2 antagonized NDRG1 expression induced by LXA4. NDRG1 siRNA suppressed viability of LPS-treated A549 cells (treatment vs. control, 0.605 ± 0.063 vs. 0.878 ± 0.083, P = 0.040) and ENaC-α expression (treatment vs. control, 0.458 ± 0.038 vs. 0.711 ± 0.035, P = 0.008). LY294002 inhibited NDRG1 (treatment vs. control, 0.459 ± 0.023 vs. 0.726 ± 0.020, P = 0.001) and ENaC-α (treatment vs. control, 0.236 ± 0.021 vs. 0.814 ± 0.025, P < 0.001) expressions and serum- and glucocorticoid-inducible kinase 1 phosphorylation (treatment vs. control, 0.442 ± 0.024 vs. 1.046 ± 0.082, P = 0.002), indicating the PI3K signaling pathway was involved in regulating NDRG1 expression induced by LXA4.

ConclusionOur research uncovered a critical role of NDRG1 in LXA4 alleviation of LPS-induced A549 cell injury through mediating PI3K signaling to restore ENaC expression.

A549 Cells ; Acute Lung Injury ; metabolism ; Cell Cycle Proteins ; metabolism ; Cell Line ; Epithelial Sodium Channels ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Lipopolysaccharides ; pharmacology ; Lipoxins ; pharmacology ; Signal Transduction ; drug effects

Objective To identify the epidemic characteristics and risk factors of an emerging infectious disease-severe fever with thrombocytopenia syndrome (SFTS) in Hubei province.Methods Active surveillance program on SFTS was set up in monitoring sites-hospitals,at the township level or above,in Suizhou,Huanggang and Wuhan from January to December,2010.Specific surveillance program on SFTS was launched across the province in hospitals above the county level.Cases that matched the definition of surveillance case were identified and reported to Centers for Disease Control and Prevention (CDCs).Cases were interviewed and their blood samples collected and detected using PCR and virus isolation.We also conducted serum antibody surveys among healthy population and livestock and surveillance on vector ticks in those high-epidemic areas.Results 188 cases that matched the definition of surveillance case and 21 deaths were reported in 11 cities,32 countries and 100 towns in 2010,with an incidence rate of 0.33/106.The fatality rate was 11.2％.Data showed that the patients were from hilly areas at the altitude elevated between 28-940 meters.The epidemic period was between April and December with the peak from May to September.The youngest case was an 11-year old,while the eldest was 81 with median age as 56-year old.95.3 ％ of the patients were farmers.All Patients did not have the history of traveling,two weeks before the onset of SFTS.93.6％ of the patients engaged in different kind of work which was associated with agriculture.52.8％ of the patients had been exposed to ticks.22.0％ of the patients had been bitten by ticks.Skin injury was found in 64.2％ of the patients.Samples from 129 cases (68.6％) were collected and detected,with 67.4％ of them (87 cases) showed positive by Real time-PCR for SFTS virus.An elevation in antibody titer by a factor of four or evidence of sero-conversion was observed in 11 patients; SFTS virus was isolated from 2 patients.The total antibody positive rates were 3.8％,55.0％ (6/11 ),36.7％ (2/3) and 80.0％ (4/5) respectively in healthy population,dogs,sheep and cows.Ticks from grass,cattle and sheep were detected positive by Real time-PCR.Conclusion Most cases of SFTS in Hubei were infected by SFTS virus,and cases of livestock were infected by SFTS virus.Ticks might serve as an important vector.Skin injury,exposure to tick bites seemed to be the risk factors.

Objective According to results from the two-month consecutive surveillance program in Maanshan,six suspected cases of non-O1 non-O139 Vibrio (V.) cholerae infection,were found that called for identification of pathogens as well as molecular-epidemiological analysis to determine the aggregation of the epidemic situation.Methods Biochemical and serotype identification,hemolysis test,and drug sensitive test were used to detect the drug resistance spectrum.Real-time PCR and conventional PCR were used to detect the presence of V.cholerae specific genes,virulent genes and its related genes,including ompW,ctx,tcpA,toxR,hlyA,zot,ace,rstR and g ⅢCTX.Pulsed-field gel electrophoresis (PFGE) was used to analyze the molecular type of strains.Results All the six isolates of non-O 1 non-O 139 V.cholerae were identified by biochemical and serologic tests,and appeared to be β hemolytic.Twelve out of the 14 kinds of drugs showed 100％ sensitive.All isolates were positive of ompW gene by real-time PCR,but negative for ctx,tcpA,zot,ace,rstR and gⅢ CTK.Five of the six isolates were positive for toxR and hlyA,except for strain 1001434446.All strains had different PFGE types,but two strains had similar types.All strains had a low similarity compared to the toxigenic V.cholerae.Conclusion Six cases ofnon-O1 and non-O139 nontoxigenic V.cholerae infection appeared in the same period.Along with epide(m)iological information,we noticed that these cases had a sporadic nature,but frequently appeared in the same area.We got the impression that public health measurements should be strengthened,with special attention paid to those diarrhea outbreaks caused by non-O 1 /non-O 139 strains since V.cholerae had appeared in low incidence.

Objective To investigate the characteristic of subtypes and genetic diversity of HIV-1 circulating in Hubei province and its molecular epidemiological linkages with regard to risk factors of viral transmission.Methods plasma samples of 80 diagnosed individuals was characterized.The gene fragments of gag were amplified by reverse transcriptase polymerase chain reaction (RT-PCR) and HIV-1 genotypes were determined based on the nucleotide sequences of gag region.Results Seven HIV-1 group M subtypes or CRF including B,B',G,CRF01-AE,CRF07-BC,CRF08-BC and CRF15-01B were identified.CRF01-AE was found to be the most dominant subtype (48.4％) followed by CRF7-BC (22.6％) and B' (12.9％).Conclusion The data from this study indicate the existence of multiple HIV-1 subtypes or CRFs in Hubei province and the surveillance of HIV-1 gene variation should be paid more attention to.

BACKGROUNDTumor necrosis factor-alpha (TNF-α) is a pleiotropic proinflammatory cytokine and contributes to many kinds of cardiovascular diseases via its receptors (TNFR1/TNFR2). We hypothesize that TNF-α plays a role in the pathogenesis of chronic atrial fibrillation (AF).

METHODSSixty-seven consecutive patients who were scheduled to have cardiac surgery were enrolled into the study. Thirty-one patients with rheumatic heart disease (RHD) and AF were enrolled as study group (AF group). The sinus rhythm (SR) control groups consisted of 20 patients with RHD and 16 patients with coronary artery disease (CAD). Peripheral blood sample was collected before the operation. About 5 mm(3) left atrial tissue was disserted during the operation and was separated into three parts for Western blotting, real time polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) analysis.

RESULTSCompared with the controls (RHD SR and CAD SR), the levels of TNF-α ((14.40 ± 5.45) pg/ml vs. (4.20 ± 3.19) pg/ml vs. (2.68 ± 2.20) pg/ml, P = 0.000) and its soluble receptor 1 (sTNFR1) ((1623.9 ± 558.6) pg/ml vs. (1222.3 ± 175.6) pg/ml vs. (1387.5 ± 362.2) pg/ml, P = 0.001) in plasma were higher in patients with AF. TNF-α level had positive correlation with the left atrial diameter (LAD) (r = 0.642, P = 0.000). Western blotting analysis showed that the protein levels of TNF-α (0.618 ± 0.236 vs. 0.234 ± 0.178 vs. 0.180 ± 0.103, P = 0.000) were higher in patients with AF. The RT-PCR analysis results demonstrated that the mRNA expression of TNF-α (0.103 ± 0.047 vs. 0.031 ± 0.027 vs. 0.023 ± 0.018, P = 0.000) increased in patients with AF. IHC analysis displayed that, comparing to the SR, the expression of TNF-α (0.125 ± 0.025 vs. 0.080 ± 0.027 vs. 0.070 ± 0.023, P = 0.000) increased in the AF group. The protein level and mRNA expression of TNF-α also had positive correlation with left atrium diameter (LAD) (r = 0.415, P = 0.000 and r = 0.499, P = 0.000).

CONCLUSIONSThe results revealed that TNF-α elevated in the plasma and left atrial tissue and had positive correlation with LAD in patients of chronic AF. TNF-α might involve in the pathogenesis of chronic AF.

Aged ; Atrial Fibrillation ; blood ; metabolism ; Blotting, Western ; Female ; Heart Atria ; metabolism ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Real-Time Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; blood ; genetics ; metabolism

Objective The purpose of this study is to evaluate the in-hospital clinical outcome of patients with coronary artery disease who underwent transradial intervention (TRI) and analyze the predictors of chinical outcome. Methods From May 2004 to May 2009, there were 16 281 patients who underwent transradial intervention, as well as 5388 patients who underwent transfemoral intervention (TFI) at our institution. The clinical characteristics, procedural characteristics, and in-hospital clinical adverse events were compared between TRI and TFI groups. Multivariable logistic regression analysis was performed to determine predictors of in-hospital major adverse cardiac events ( composite of death, myocardial infarction,or target lesion revascularization) of TRI. Results The annulations time was significantly longer for TRIthan TFI (P ＜0. 01 ), fluoroscopy time, amount of contrast agent and procedural success rate (95.5% for TRI and 96. 2% for TFI) were similar between the two groups. However, the rates of vascular complications (0. 1% for TRI group and 1.3% for TFI group, P ＜0. 01 ), incidence of in-hospital major adverse cardiac events (1.6% vs. 3. 8%, P＜ 0.01) and in-hospital death (0.2% vs. 0.4%, P＜0.01) were all significantly lower in TRI group compared with TFI group. The following characteristics were identified as independent multivariate predictors of in-hospital major adverse cardiac events of TRI: age ≥65 ( OR: 1.98,95% CI: 1. 50 - 2. 61, P ＜ 0. 01 ), prior myocardial infarction ( OR:2. 14, 95% CI: 1.63 - 2. 82, P ＜0. 01 ), use of drug-eluting stent (DES) ( OR:0. 68, 95% CI:0. 47 - 0. 98, P = 0. 04 ), dissection during procedure (OR:4.08, 95%CI:2.28-7.33, P＜0.01), left main lesion (OR:2. 12, 95% CI:1.09-4. 13, P=0.03), number of implanted stents (OR:1.25, 95% CI:1.09 - 1.43, P ＜0.01), and total stented length (OR:1.01, 95% CI:1. 00 -1. 02 , P=0.03). Conclusions In this large single-centre patient cohort, the transradial intervention is superior to transfemoral intervention in terms of in-hospital safety and efficacy. Age ≥ 65, prior myocardial infarction, use of DES, dissection during procedure, left main lesion, number of implanted stents and total stented length were identified as independent multivariate predictors of in-hospital major adverse cardiac events of TRI.

Objective To explore multiple slices computed tomography (MSCT) and magnetic resonance imaging ( MRI) features of duplication of the internal auditory canal ( DIAC) in order to improve the accuracy of diagnosis. Methods Four cases(5 ears) were analyzed and the related documents were reviewed retrospectively. MSCT was performed on all cases, and two cases had MRI scanning at the same time. Results MSCT has shown that the internal auditory canal were divided into two canals by a bony septum in 5 ears. The superior canal ended in a very narrow connection to the facial canal, the inferior portion ended in connection to the cochlea and vestibule. The bony septums from the 2 ears were found no longer intact. The sum of diameter of the two canals was greater than 2 mm. In addition, S ears were found to have an enlarged vestibules and the hypoplasia lateral semicircular canals, and meanwhile, 2 ears of them were combined with ipsilateral microtia. Also 1 case of them was combined with microtia, outer acoustic atresia as well as abnormal middle ear. Multiplanar reconstruction and volume rendering images can entirely show the bony septum and two canals. In this study, the vestibular nerve, cochlear nerve and facial nerve were total hypoplastic in one ear, in the other ear, the vestibular and cochlear nerve were hypoplastic, and however, the facial nerve was intact. Conclusions MSCT can clearly depict duplaication of the internal auditory canals and concomitant anomalies. MRI can clearly show the neural components and their associated malformation.