2.Identification of Bletillae Rhizoma and its adulterants by SNPs in ITS2.
Dan ZHAO ; Tao ZHOU ; Wei-ke JIANG ; Cheng-hong XIAO ; Chuan-zhi KANG
China Journal of Chinese Materia Medica 2015;40(18):3573-3578
To establish a molecular identification method for Bletillae Rhizoma, this paper extracted genome DNA from Bletillae Rhizoma and its adulterants. The sequences of rDNA ITS2 were sequenced after amplifying. Then multiple alignments of ITS2 were constructed phylogenetic tree with Neighbor Joining by MEGA 5. 1 and found out SNPs loci. The result showed that rDNA ITS2 region could identify Bletillae Rhizoma and its adulterants. There existed the SNPs loci, which could identify Bletilla striata and B. ochracea. Furthermore, we designed specific primers against the SNPs loci of B. striata and B. ochracea, then screened primers and optimized the PCR amplification conditions. Finally, the DNA of B. striata and B. ochracea were specifically amplified by BJ59-412F, BJ59-412R and HHBJ-225R. The length of amplification products were respectively about 350 bp and 520 bp that were effectively identified of B. striata and B. ochracea. While, the adulterants of Bletillae Rhizoma were no-reaction occurring. To sum up, the amplification conditions of the primers can identify B. striata, B. ochracea and their adulterants successfully at the same time. This method was easy, time-saving, and reliable, which can be used as a rapid method for molecular identification of Bletillae Rhizoma.
Base Sequence
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DNA Primers
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genetics
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DNA, Intergenic
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genetics
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DNA, Plant
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genetics
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Drug Contamination
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prevention & control
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Molecular Sequence Data
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Orchidaceae
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classification
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genetics
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Phylogeny
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Polymorphism, Single Nucleotide
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Rhizome
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classification
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genetics
3.Rapid molecular identification of pseudostellariae radix.
Dan ZHAO ; Tao ZHOU ; Wei-Ke JIANG ; Yuan YUAN ; Cheng-Hong XIAO ; Wei ZHENG
China Journal of Chinese Materia Medica 2014;39(19):3689-3694
To establish a convenient and rapid method for identification of Pseudostellariae Radix by molecular identification, the rDNA-ITS sequences of Pseudostella riaheterophylla and its adulterants had been aligned to find out specific fragment. The specific primers against the fragment were designed and the PCR amplification conditions were optimized. The fluorescence reaction of the PCR products colored by 100 x SYBR Green I was observed under UV. The concentration of reaction buffer included 5.5 μL DNA Taq polymerase premix, 10 pmol Tzs-2F and 10 pmol Tzs-2R, 20-80 ng template DNA, and plus double sterile distilled water to 25 μL. The PCR thermal profile was as follows: predenaturation at 95 degrees C for 1 min, followed by 30 cycles of denaturation at 95 degrees C for 5 seconds, primer annealing and extension at 56 degrees C for 15 seconds, then it was extension at 72 degrees C for 30 seconds. The fluorescence reaction of Pseudostellariae Radix showed green fluorescence, while adulterants had not. Extraction, amplification DNA and all steps of molecular identification could be completed successfully in 40 minutes. The approach could amplify DNA template of Pseudostellariae Radix specificity, and its product with 1 μL 100 x SYBR Green I could engender green fluorescence under UV. The method was simple and accurate, so it could be used for identification of Chinese traditional medicine.
Caryophyllaceae
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classification
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genetics
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DNA Primers
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genetics
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Drug Contamination
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prevention & control
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Plant Roots
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classification
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genetics
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Polymerase Chain Reaction
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methods
4.Study on seed quality test and quality standard of Pesudostellaria heterophylla.
Cheng-Hong XIAO ; Tao ZHOU ; Wei-Ke JIANG ; Min CHEN ; Hou-Xi XIONG ; Ming-Wu LIAO
China Journal of Chinese Materia Medica 2014;39(16):3042-3047
Referring to the rules for agricultural seed testing (GB /T 3543-1995) issued by China, the test of sampling, seed purity, weight per 1 000 seeds, seed moisture, seed viability and germination rate had been studied for screening seed quality test methods of Pesudostellaria heterophylla. The seed quality from different collection areas was measured. The results showed that at least 6.5 g seeds should be sampled and passed through 10-mesh sieve for purity analysis. The weight of 1 000 seeds was determined by using the 500-seed method. The phenotypic observation and size measurement were used for authenticity testing. The seed moisture was determined under the higher temperature (130 ± 2) degrees C for 5 hours. The seeds were dipped into 0.2% TTC sustaining 1 hour at 40 degrees C, then the viability could be determined. The break dormancy seeds were cultured on sand at 10 degrees C. K cluster analysis was applied for the data analysis, the seed quality from different collection areas grading of P. Heterophylla was described as three grades. The seed quality of each grade should reach following requirements: for first grade seeds, germination rate ≥ 86%, 1 000-grain weight ≥ 2.59 g, purity ≥ 87%, moisture ≤ 13.1%; for second grade seeds, germination rate ≥ 70%, 1 000-grain weight ≥ 2.40 g, purity ≥ 77%, moisture ≤ 14.3%; for third grade seeds, germination rate ≥ 41%, 1 000-grain weight ≥ 2.29 g, purity ≥ 76%, moisture ≤ 15.8%. The seed testing methods for quality items of P. heterophylla had been initially established, as well as the primary P. heterophylla seed quality classification standard.
Caryophyllaceae
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chemistry
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growth & development
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Germination
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Quality Control
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Seeds
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chemistry
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growth & development
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Water
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analysis
5.Arterial spin labeling analysis of the cerebral blood flow in systemic lupus erythematosus patients
Xiaolu JIANG ; Zhen CHENG ; Longjiang ZHANG ; Yan ZHOU ; Jun KE ; Song LUO ; Gang ZHENG ; Zongjun ZHANG
Journal of Medical Postgraduates 2016;29(5):495-499
Objective Neuropsychiatric systemic lupus erythematosus ( SLE) is a common complication of SLE, whose path-ogenesis is not yet clear but associated with the alteration of cerebral blood flow ( CBF) in some studies.This study was to investigate the CBF alteration in SLE patients without overt neuropsychiatric symptoms by arterial spin labeling ( ASL) MRI. Methods Twenty-eight SLE patients without overt neuropsychiatric symptoms and 30 age-and sex-matched healthy controls underwent conventional MRI and ASL examinations, and all received such neuropsychologic tests as number connecting test-A ( NCT-A ) , digit symbol test ( DST ) , self-rating anxiety scale ( SAS ) , and self-rating depression scale ( SDS) .Independent sample-t test was used to detect the mean CBF in the whole brain, gray matter, and white matter of the SLE patients and healthy controls.The voxel-wise CBF maps of the two groups of subjects were further analyzed with the SPM8 software to compare the regional CBF between the two groups, followed by evaluation of the correlation between the regional CBF values and clinical markers. Results In comparison with the healthy controls, the SLE pa-tients showed significantly reduced CBF in the gray matter (40.5 ±3.7 vs 37.3 ±6.5, P=0.028) and the whole brain (38.0 ±3.5 vs 35.1 ±6.1, P=0.032), especially in the supplementary motor area and the adjacent middle cingulate, anterior cingulate, left medial frontal gyrus, left inferior frontal gyrus, and left insula (P<0.05, FWE corrected).The NCT-A score was negatively correlated with the CBF values of the left medial frontal gyrus (r=-0.402, P=0.032) and left inferior frontal gyrus (r=-0.382, P=0.045) of the SLE patients. Conclusion ASL and MRI showed significantly reduced cerebral blood flow in the SLE patient without overt neu-ropsychiatric manifestations, which was correlated with the change of the patient's cognitive function.
6.Predictive value of neutrophil to lymphocyte ratio for inhospital mortality in type 2 diabetic patients with acute myocardial infarction
Li KE ; Bei CHENG ; Benling QI ; Tangmeng GUO ; Min ZHOU ; Bin YANG ; Lulu CHEN
Chinese Journal of Endocrinology and Metabolism 2017;33(6):479-484
Objective To explore the prognostic value of biomarkers in type 2 diabetic patients with acute myocardial infarction (AMI), this study was to investigate the associations between the neutrophil to lymphocyte ratio (NLR), the Global Registry of Acute Coronary Events (GRACE) score and in-hospital mortality. MethodsSeven hundred and seven consecutive AMI patients were divided into diabetic group (DM-AMI group), impaired glucose tolerance group (IGT-AMI group), and normal glycemic group (NGT-AMI group). The laboratory and clinical characteristics were assessed retrospectively from the medical records. The NLR and GRACE score were calculated. Results In AMI patients, the DM-AMI group had significantly higher NLR and GRACE scores compared with those from the IGT-AMI group and NGT-AMI group (P<0.01 or P<0.05). In DM-AMI group, the NLR and GRACE score were considerably elevated in the elderly DM-AMI group compared with their younger counterparts (both P<0.01). Furthermore, the NLR was considerably higher in the high-risk group than those in both the low- and medium-risk groups based on the GRACE score (both P<0.01). The NLR was positively correlated with the GRACE score in DM-AMI group(r=0.425, P<0.01). The NLR level and GRACE score were higher in the death group than those in surviving patients (both P<0.01). The optimal cut-off levels of 9.36 for NLR and 166 for GRACE score seem to predicte death in-hospital. Based on the receiver operating characteristic curve, when to predict death in-hospital, the best cutoff value of NLR was 9.36 (sensitivity 80.8%, specificity 69.6%; area under curve 0.787), and the best cutoff value of GRACE score was 166 (sensitivity 76.9%, specificity 76.4%; area under curve 0.778). Conclusion An elevated NLR is a potential predictor of in-hospital mortality in type 2 diabetic patients with AMI, which could help clinicians indentify high-risk patients and determine appropriate treatment strategies. <英文关鍵词>>=Neutrophil to lymphocyte ratio; In-hospital mortality; Acute myocardial infarction; Diabetes mellitus, type 2
7.Analysis on present situation of dental instruments cleaning and disinfection in Hainan province
Shuping ZHOU ; Chenyun XU ; Yajuan KE ; Shuming XIANYU ; Linqiu FU ; Wei FU ; Xiuju WANG ; Cheng SUN
Chongqing Medicine 2014;(10):1231-1233
Objective To analyze the present situation of the dental instrument cleaning and disinfection in Hainan province and to explore the management mode of dental instrument cleaning and disinfection which is suitable for the provincial situation .Meth-ods By adopting the stratified random sampling method according to the hospital grades ,24 hospitals in Hainan province were per-formed the questionnaire survey on the cleaning mode of oral instruments ,layout of cleaning and disinfection room ,cleaning method and facilities ,protection measures and training of cleaning staff .Results 14 hospitals (58 .33% ) had the sterilization and supply center for conducting the centralized processing on the dental instruments .The tertiary hospitals and the second-grade hospitals had the independent cleaning and disinfection rooms with the rational layout and professional cleaning staff ;the safeguard facilities had the application in place ,the training of the related cleaning and disinfection work and the cleaning process conformed the require-ment of the standards .Among 10 first-grade and below hospitals ,only 1 hospital(10 .00% ) had the rational layout of cleaning and disinfection rooms ;3 hospitals(30 .00% ) had the professional cleaning staff ;the related training of the cleaning staff was not basi-cally carried out and the safeguard was not in place ,most of the cleaning and disinfection instruments and the cleaning process were not in accordance with the requirements .The qualification rates of instruments cleaning and disinfection in different grades of hospi-tals by the ATP bioluminescence assay were 100 .00% ,90 .00% and 80 .00% .Conclusion The existing problems are general and prominent in the hospitals of the first-grade and below .It is suggested that the regionalized disinfection and supply management mode is implemented for maximally realizing the optimized resource configuration in the disinfection and supply center .
8.Comparative study on effect of osthole and genistein on peak bone mass in rats.
Kui CHENG ; Bao-Feng GE ; Ping ZHEN ; Ke-Ming CHEN ; Xiao-Ni MA ; Jian ZHOU ; Peng SONG ; Hui-Ping MA
China Journal of Orthopaedics and Traumatology 2014;27(7):587-591
OBJECTIVETo compare the ability of osthole (OST) and genistein (GEN) in enhancing bone peak bone mass of rats to prevent osteoporosis.
METHODSThirty-six female one-month-old SD rats of (125 +/- 3) g body weight were randomly divided into three groups, 12 rats in each group, one group was orally administered osthole at 9 mg x kg(-1) d(-1), one group was given genistein at 10 mg x kg(-1) d(-1) and another was given equal quantity of distilled water as the control. The body weight was monitored weekly and the bone mineral density (BMD) of total body was measured every month. All rats were sacrificed after three months, the femoral bone mineral density, the serum levels of osteocalcin (OC) and anti-tartaric acid phosphatase 5b (TRACP 5b) were measured by Elisa. The bone microarchitectures were analyzed with micro-CT and the bone biomechanics properties were tested with universal material machine.
RESULTSNo significant differences were observed between O-treated or GEN group and the control for the food-intake and body weight during three months. However, the rats treated with OST had significant higher BMD for both total body and femur than the control and GEN group. The O-treated rats also had higher level of serum OC and lower level of TRACP 5b. Besides, they owned bigger bone volume/tissue volume, trabecular thickness, trabecular number but smaller trabecular spacing. In the three point bending tests of femurs,they were found to have larger maximum load, the young's modulus and structural model index (SMI).
CONCLUSIONOrally administered osthole could efficiently increase the peak bone mass of rats,which provide new ideas for preventing osteoporosis.
Acid Phosphatase ; blood ; Animals ; Body Weight ; drug effects ; Bone Density ; drug effects ; Coumarins ; pharmacology ; Female ; Femur ; diagnostic imaging ; drug effects ; pathology ; Genistein ; pharmacology ; Isoenzymes ; blood ; Osteocalcin ; blood ; Radiography ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase
9.Full fascia closure with interrupted absorbable suture and layered closure with interrupted silk suture in abdominal incision:comparison of curative effects and biocompatibility
Dan LI ; Jing ZHUANG ; Yonggang LIU ; Hao ZHOU ; Kaixuan CHEN ; Ke CHENG ; Jinbang WANG ; Baodong LI ; Suxia LUO ; Guangsen HAN
Chinese Journal of Tissue Engineering Research 2014;(43):6996-7000
BACKGROUND:Abdominal incision healing is not only related with the patient’s own situation, but also closely related with the surgeon's suture technique, suture method, choice of stitches. <br> OBJECTIVE:To compare the absorbable sutures and silk sutures for abdominal incision. <br> METHODS:Total y 153 colorectal cancer patients, including 91 males and 62 females, aged 30-82 years, were randomly divided into observation group (n=78) and control group (n=75). An abdominal midline incision was made in al patients receiving radical surgery of colorectal cancer. The Vicryl suture and silk suture were respectively used in the observation and control groups for abdominal incision closure. Suturing time, length of hospital stay, incision infection, disruption of wound, fat liquefaction of wound and rejection were compared between two groups. <br> RESULTS AND CONCLUSION:The suturing time and length of hospital stay were less in the observation group than the control group (P<0.05). In the observation group, there were three cases of incision infection, but no incision dehiscence and rejection occurred;in the control group, there were 10 cases of incision infection, 4 cases of incision dehiscence, and 5 cases of rejection. A significant difference was found in the incision infection, dehiscence and rejection between the two groups (P<0.05). Hospitalization expenses and fat liquefaction of incision had no difference between the two groups. these findings indicate that the Vicryl plus as an absorbable suture is simple, effective and safe that can promote wound healing and reduce complications.
10.Role of a disintegrin and metalloprotease with thrombospondin type 1 motifs in aged rat myocardium after ischemic preconditioning
Yong WANG ; Congxin HUANG ; Jinsong CHENG ; Yifeng ZHOU ; Hui WANG ; Wenjing WU ; Wenqiang LIAO ; Jianyan WEN ; Yuannan KE ; Jingang ZHENG
Chinese Journal of Geriatrics 2011;30(1):54-58
Objective To investigate the effect of ischemic preconditioning (IPC) on the expression of a disintegrin and metalloprotease with thrombospondin type 1 motifs (ADAMTS-1), and to study whether the application of small interfering (si)RNA specifically targeting ADAMTS-1 would help to recover IPC protection in the aged heart. Methods The 32 young (4 months) and 32 aged(24 months) male Sprague-Dawley (SD) rats were assigned randomly to IPC group (n=20) and sham operated group (n= 12) respectively. Myocardial samples from the ischemic-reperfused region were harvested for detecting the ADAMTS-1 expression. In addition, the 110 aged SD rats were assignedrandomly to ADAMTS-1 siRNA group and control group (n=55, each). The effects of ADAMTS-1siRNA transfcction on the expression of ADAMTS-1 protein, myocardial infarction survival rate,heart function and myocardial infarction size after IPC were observed.Results Twenty-four hours after IPC, the ADAMTS-1 protein expression increased significantly in iscbemic-reperfused region both in young and aged rats (P<0. 05), and the protein expression was higher in aged rats than in young rats (P<0.05). In young-IPC group, the absorbency showed ADAMTS-1 protein expression at 0 hrs and 24 hrs after IPC were 0. 05±0.01 and 0.12±0.03 by immunohistochemical staining, and were 0.68±0. 16 and 1. 17±0.21 by Western blots respectively. In aged-IPC group, the absorbency showed ADAMTS-1 protein expression at 0 hrs and 24 hrs after IPC were 0.07±0. 03 and 0.21 ±0.04 by immunohistochemical staining, and were 0. 76±0. 21 and 1. 48±0. 17 by Western blots. In the aged rats, ADAMTS-1 siRNA transfection inhibited ADAMTS-1 protein expression (0. 66±0. 19and 0.78±0.21, by Western blots at 0 hrs and 24 hrs after IPC, P>0.05), but didn't improve myocardial infarction survival rates [ADAMTS-1 siRNA group and sham operated group: 14.3% (5/35) vs. 17.1 %(6/35), P>0.05], left ventricular fractional shortening [(14.0±3.2)% vs. (13.0±2.9)%, P>0.05] and myocardial infarction size[(39.0±4.1)% vs. (38.0±5.3)%, P>0.05].Conclusions ADAMTS-1 expression induced by IPC increases significantly in aged versus in young rats. ADAMTS-1 knockdown by siRNA inhibits ADAMTS-1 protein expression but cannot recover the age-associated loss of IPC protection.