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ObjectiveTo analyze the sequence variation and genetic diversity of 47 Isatis indigotica germplasm materials， and carry out the study on the genetic differentiation and structure. MethodGenomic DNA of 47 I. indigotica germplasm materials were extracted by kit extraction method. Two chloroplast DNA （cp DNA） sequences and five inter-simple sequence repeat （ISSR） primers were used for amplification and sequencing. Chromas， Mega 7.0， DanSP5， and GenALEx were used to calibrate， splice， and analyze the sequence characteristics. PERMUT and PopGen 1.31 were used to analyze the genetic diversity parameters and genetic structure， and NTSYS was used to obtain the unweighted pair-group method with arithmetic means（UPGMA） clustering tree plot of 47 I. indigotica germplasm materials. ResultA total of 129 samples from 47 I. indigotica germplasm materials were successfully amplified and sequenced. The length of 2 cp DNA sequences after spliced was 1 412 bp， and there were 377 polymorphic variation loci， and 36 haplotypes. Fu and Li's D* test was significant （P<0.01）. The values of Pi， HS， and HT based on cp DNA were 0.119 89， 0.787， and 0.891， respectively. The genetic differentiation coefficients of gene differentiation coefficient（Gst）， nucleotide differentiation coefficient（Nst）， and fixation index（Fst） were 0.117， 0.468， and 0.488， respectively， and the gene flow （Nm） was 0.615. The mean values of PPB， Shannon information diversity index（I）， Nei's genetic diversity index（H）， and Gst based on ISSR were 78.85%， 0.334 8， 0.218 6， and 0.754 4， respectively， and the Nm value was 0.162 8. ConclusionI. indigotica has high genetic diversity and abundant haplotypes at the species level， with abundant haplotypes. Genetic differentiation among different germplasm materials is obvious， and gene exchange is not frequent. Genetic variation mainly exists among populations. The population has accumulated various low-frequency gene mutations recently， suggesting that it has experienced significant regional expansion in the history.
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ObjectiveTo analyze the sequence variation and genetic diversity of 47 Isatis indigotica germplasm materials， and carry out the study on the genetic differentiation and structure. MethodGenomic DNA of 47 I. indigotica germplasm materials were extracted by kit extraction method. Two chloroplast DNA （cp DNA） sequences and five inter-simple sequence repeat （ISSR） primers were used for amplification and sequencing. Chromas， Mega 7.0， DanSP5， and GenALEx were used to calibrate， splice， and analyze the sequence characteristics. PERMUT and PopGen 1.31 were used to analyze the genetic diversity parameters and genetic structure， and NTSYS was used to obtain the unweighted pair-group method with arithmetic means（UPGMA） clustering tree plot of 47 I. indigotica germplasm materials. ResultA total of 129 samples from 47 I. indigotica germplasm materials were successfully amplified and sequenced. The length of 2 cp DNA sequences after spliced was 1 412 bp， and there were 377 polymorphic variation loci， and 36 haplotypes. Fu and Li's D* test was significant （P<0.01）. The values of Pi， HS， and HT based on cp DNA were 0.119 89， 0.787， and 0.891， respectively. The genetic differentiation coefficients of gene differentiation coefficient（Gst）， nucleotide differentiation coefficient（Nst）， and fixation index（Fst） were 0.117， 0.468， and 0.488， respectively， and the gene flow （Nm） was 0.615. The mean values of PPB， Shannon information diversity index（I）， Nei's genetic diversity index（H）， and Gst based on ISSR were 78.85%， 0.334 8， 0.218 6， and 0.754 4， respectively， and the Nm value was 0.162 8. ConclusionI. indigotica has high genetic diversity and abundant haplotypes at the species level， with abundant haplotypes. Genetic differentiation among different germplasm materials is obvious， and gene exchange is not frequent. Genetic variation mainly exists among populations. The population has accumulated various low-frequency gene mutations recently， suggesting that it has experienced significant regional expansion in the history.
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Objective:The biological mechanism of <italic>Codonopsis pilosula</italic> adaptation to drought was explored by determining the root metabolome of <italic>C. pilosula</italic> during harvesting. Method:Non-targeted metabonomics LC-MS was used to screen differential metabolites by multivariate statistical analysis，univariate statistical analysis and metabolic pathway enrichment analysis. Result:①There were 274 metabolites in LD vs CK group，142 of which were up-regulated and 132 of which were down-regulated. There were 284 metabolites with significant difference in MD vs CK group，of which 157 were up-regulated and 127 were down-regulated. There were 317 metabolites with significant difference in SD vs CK group，of which 133 were up-regulated and 184 were down-regulated. ②Differential metabolites were annotated into kyoto Encyclopedia of Gene and Genomes （KEGG） database and 82 differential metabolic pathways were obtained，among which sphingolipids metabolism was significantly enriched （<italic>P</italic><0.01）.Metabolism of arginine and proline，tryptophan，alanine，galactose，nicotinic acid and nicotinamide，cysteine and methionine，arachidonic acid，linolenic acid and glycerides were significantly enriched in different metabolite pathways （<italic>P</italic><0.05）. ③The metabolites of the three comparison groups before and after enrichment were classified and analyzed. It was found that they were mainly concentrated in fatty acyls group，carboxylic acid and derivatives，and organ oxygen compounds，followed by sphingolipids，indoles and derivatives，organonitrogen compounds，glycerophospholipids，pyridines and derivatives，peptidomimetics，glycerolipids and so on.In the drought stress of <italic>C. codonopsis</italic>，carbohydrate related metabolites were mainly up-regulated，lipid related metabolites were mainly down-regulated，and all other metabolites were up-regulated. Conclusion:The changes of metabolites in the roots of <italic>C. pilosula</italic> under drought stress were elucidated. carbohydrate and lipid-related metabolites were the main products of <italic>C. pilosula</italic> under drought stress，and these metabolites may be the main reason to improve the ability of <italic>C. pilosula</italic> to resist drought，which laid a foundation for further study on the mechanism of <italic>C. pilosula</italic> to resist drought.
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Objective: To observe the clinical efficacy of tuina at head and abdomen for headache due to phlegm turbidity. Methods:A total of 56 patients with headache due to phlegm turbidity were randomized into a tuina group and a Chinese medicine group by the random number table, with 28 cases in each group. The tuina group was treated mainly with tuina at the head and abdomen, while the Chinese medicine group was treated with oral administration of Ban Xia Bai Zhu Tian Ma Tang(Pinellia, Atractylodes Macrocephala and GastrodiaDecoction). The course of treatment was 30 d. The scores of headache index, traditional Chinese medicine syndrome scale, and the therapeutic efficacy were observed. Results:There were 2 dropouts in each group during treatment. The total effective rate was 92.3％ in the tuina group, significantly higher than 76.9％ in the Chinese medicine group (P<0.05). The scores of headache index and traditional Chinese medicine syndrome scale in both groups decreased after treatment (bothP<0.05), and scores in the tuina group were lower than those in the Chinese medicine group (bothP<0.05). Conclusion:Tuina mainly at head and abdomen is effective in treating headache due to phlegm turbidity, and has a better effect than Ban Xia Bai Zhu Tian Ma Tang (Pinellia, Atractylodes Macrocephala and GastrodiaDecoction).
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OBJECTIVE@#To investigate anti-inflammatory and immunomodulation effects of different ecotype from Isatidis Radix growing in Gansu province.@*METHODS@#Mice were randomly divided into 6 groups (=11)and used the auricular swelling and paw edema to observe the anti-inflammatory effects of Isatidis Radix; Mice were randomly divided into 7 groups (=11) and through the gasbag synovitis model to observe the anti-inflammatory effects of Isatidis Radix; Mice were randomly divided into 6 groups (=11), the immunosuppressed model were established by injection of cyclophosphamide (CTX) to study the effects of Isatidis Radix on index of thymus, blood routine and cytokines.@*RESULTS@#Gansu different ecotype from Isatidis Radix could reduce the swelling of the mice auricle, paw edema and total protein, leukotriene B(LTB)and malonaldehyde(MDA) in airbag synovitis exudates, and upgrade serum levels of superoxide dismutase (SOD); Degrade the tumor necrosis factor-α (TNF-α) and upgrade the index of thymus, the number of red and white corpuscles, the level of interferon-γ (IFN-γ), interleukin-4 (IL-4) (<0.05, 0.01) of mice immunosuppressed model; Above the research of anti-inflammatory and immunomodulation, there were no significant differences between Isatidis Radix of Gansu different ecotype and tetraploid.@*CONCLUSIONS@#Different ecotype of Isatidis Radix has obvious functions in anti-inflammatory and immunomodulation, but there are no significant differences between Gansu different ecotype and tetraploid.
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Animals , Mice , Anti-Inflammatory Agents , Pharmacology , China , Cytokines , Allergy and Immunology , Drugs, Chinese Herbal , Pharmacology , Ecotype , Immunomodulation , Isatis , Chemistry , Plant Extracts , Pharmacology , Random AllocationABSTRACT
<p><b>OBJECTIVE</b>To investigate the safety and feasibility of 3D laparoscopic surgery via transrectal extraction of specimens without abdominal incision in the treatment of slow transit constipation (STC).</p><p><b>METHODS</b>From May 2015 to January 2017, 8 STC patients (6 females and 2 males) with informed consent were selected to receive subtotal colectomy with 3D laparoscopy as the no-incision incision group, in which the initial part of ascending colon and rectum were end-to-end anastomosed directly after extraction of the specimen through the rectum. Twelve STC patients (9 females and 3 males) undergoing traditional subtotal colectomy with 3D laparoscopy were selected as the traditional group by case matching method (gender, age, BMI, the difference of receiving operation time less than 12 months, same surgeon team). Perioperative parameters (operation duration, intraoperative blood loss, exhausting time, postoperative hospital stay, complications, postoperative pain score and additional pain management), inflammation index at postoperative day 1 and day 3 (leukocyte, procalcitonin, interleukin 6, C-reactive protein), postoperative peritoneal infection, wound healing, short-term and long-term efficacy, patient satisfaction evaluation (subjective hundred-mark system) at postoperative one year were compared between two groups.</p><p><b>RESULTS</b>There were no significant differences between two groups in operation duration, intraoperative blood loss, exhausting time, postoperative hospital stay and morbidity of complication (all P>0.05). Significantly lower pain scores at postoperative 6-hour (median 3.0 vs. 4.5, U=23.0, P=0.042), lower ratio of additional analgesic at postoperative day 1(1/8 vs. 7/12, P=0.040) were found in the no-incision group. Leukocyte level at postoperative day 1 was significantly lower in the no-incision group [(11.0±3.5)×10/L vs. (14.7±3.6)×10/L, t=-2.281, P=0.035]. C-reactive protein concentration at postoperative day 3 was not significantly different between two groups but with different trend [median 78.1(0.1 to 154.0) mg/L vs. 22.0 (7.0 to 55.9) mg/L,U=33.0, P=0.047]. There were no significant differences of interleukin-6 and procalcitonin between two groups(all P>0.05). All the patients had follow-up for 14-31 months. Subjective effectiveness score was 90±9 in the no-incision group and 94±6 in the traditional group without significant difference(t=-1.099, P=0.286). No long-term complications associated with abdominal infection was observed in the no-incision group.</p><p><b>CONCLUSION</b>3D laparoscopic subtotal colectomy via transrectal extraction of specimens without abdominal incision in the treatment of STC has similar short-term and long-term efficacies compared with traditional laparoscopic assisted surgery, and does not increase the probability of abdominal contamination.</p>
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Female , Humans , Male , Colectomy , Methods , Constipation , General Surgery , Laparoscopy , Length of Stay , Operative Time , Rectum , Treatment OutcomeABSTRACT
miR-214 plays a major role in the self-renewal of skin tissue. However, whether miR-214 regulates the proliferation and differentiation of human hair follicle stem cells (HFSCs) is unknown. Primary HFSCs were isolated from human scalp skin tissue, cultured, and identified using flow cytometry. An miR-214 mimic and inhibitor were constructed for transfection into HFSCs. The MTS and colony formation assays examined cell proliferation. Immunofluorescence detected the localization and expression levels of TCF4, β-catenin, and differentiation markers. Luciferase reporter and TOP/FOP Flash assays investigated whether miR-214 targeted EZH2 and regulated the Wnt/β-catenin signaling pathway. Western blot determined the expression levels of enhancer of zeste homolog 2 (EZH2), Wnt/β-catenin signaling-related proteins, and HFSC differentiation markers in cells subjected to miR-214 transfection. miR-214 expression was remarkably decreased during the proliferation and differentiation of HFSCs into transit-amplifying (TA) cells. Downregulation of miR-214 promotes the proliferation and differentiation of HFSCs. Overexpression of miR-214 led to decreased expression of EZH2, β-catenin, and TCF-4, whereas downregulation of miR-214 resulted in increased expression of EZH2, β-catenin, and TCF-4 as well as TA differentiation markers. Immunofluorescence assay revealed that inhibiting miR-214 triggered the entry of β-catenin and TCF-4 into the nucleus. The luciferase reporter and TOP/FOP Flash assays demonstrated that miR-214 directly targets EZH2 and affects Wnt/β-catenin signaling. The miR-214/EZH2/β-catenin axis could be considered a candidate target in tissue engineering and regenerative medicine for HFSCs.
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Humans , Antigens, Differentiation , Blotting, Western , Cell Proliferation , Down-Regulation , Flow Cytometry , Fluorescent Antibody Technique , Hair Follicle , Hair , In Vitro Techniques , Luciferases , Regenerative Medicine , Scalp , Skin , Stem Cells , Tissue Engineering , TransfectionABSTRACT
Objective To study the physiological characteristics of Codonopsis Radix under drought stress; To reveal the physiological mechanism of Codonopsis Radix in response to drought stress. Methods Method of pot experiment was used to set up 4 water treatments: normal water supply, mild stress, moderate stress and severe stress. Through the determination of leaf relative water content, cell membrane permeability, osmotic adjustment contents and protective enzyme activity, the effects of drought stress on physiological characteristics of Codonopsis Radix were studied. Results With the drought stress increased, the chlorophyll a, total chlorophyll content and leaf relative water content of Codonopsis Radix decreased, but chlorophyll b had no obvious change; The conductivity increased first and then decreased; MDA contents increased first and then decreased and then increased; There was no significant change in the rate of superoxide anion production; POD and CAT activity increased; SOD had no significant change; The proline content increased first and then decreased; soluble sugar decreased; soluble protein had no significant change. Conclusion Under the condition of drought stress, by increasing the content of proline to regulate cell osmotic potential, Codonopsis Radixcan increase the antioxidant enzyme activity to reduce membrane lipid peroxidation damage to cells.
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Objective To observe the clinical efficacy and safety of high-frequency diathermic therapy and intra-thoracic chemotherapy with recombinant endostar combined with cisplatin in treatment of malignant pleural effusion. Methods A total of 48 patients with malignant pleural effusion diagnosed in the First People ' s Hospital of Guiyang from September 2014 to September 2016 were randomly divided into observation group and control group with envelope method. Twenty-four patients in observation group were received high-frequency diathermic therapy and cisplation 60 mg/m2 combined with endostar 25-30 mg/m2 intra-thoracic chemotherapy. Twenty-four patients in control group were treated with high-frequency diathermic therapy and single cisplation 60 mg/m2 intra-thoracic chemotherapy. The patients were karnofsky performance scores ≥70, cooperate with intravenous systemic chemotherapy, every 21 days for a total of two or three cycles. High frequency diathermic therapy was administered twice a week for 3 weeks 30 minutes after intra-thoracic chemotherapy,treatment time of 60 minutes every time. Chi-squared test were selected to evaluate the clinical efficacy and quality of life and adverse and toxic responses, respectively. The paired samplest test were selected to evaluate the variation of tumor markers in hydrothorax of the two groups pretherapy and post-treatment. Results In 24 patients of observation group, 2 patients were complete remission (CR), 17 patients were partial remission (PR), 4 patients were disease stable (SD), 1 patient was disease progression (PD), and the objective response rate (ORR) (CR+PR) was 79.2 ％. In 24 patients of control group, 0 patient was CR, 12 patients were PR, 7 patients were SD, 5 patients were PD, and the ORR was 50.0％, there was significant difference in ORR between the two groups (χ 2 = 4.463, P = 0.035). Karnofsky performance scores in observation group was higher than that in control group after treatment, patients with clinical benefit rate was 79.2 ％ vs. 54.2 ％, but the difference in life quality between the two groups was no statistically significant (χ2=3.375, P=0.066). There was no significant difference in adverse reactions between the two groups (P>0.05). The tumor markers carcinoembryonic antigen (CEA), carbohydrate antigen 125 (CA-125), carbohydrate antigen 199 (CA-199), cytokeratin 19 fragments (CYFRA21-1) and neuron-specific enolase (NSE) in hydrothorax of the two groups were reduced by the treatment, except for the difference of CYFRA21-1 between the two group was not statistically significant (P= 0.161), the changes of the other 3 indicators in the observation group before and after treatment were greater than those in the control group (all P< 0.05). Conclusions High-frequency diathermic therapy and intra-thoracic chemotherapy with single cisplatin are effective in treating patients with malignant pleural effusion, and is more superior when combined with endostar. Additionally, the combination of the above two drugs has synergistic action and better safety, deserves to be further promoted in clinic.
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<p><b>OBJECTIVE</b>To introduce the use of a self-made specimen protective sleeve in laparoscopic resection for upper or mid rectal cancer and sigmoid colon cancer with transrectal specimen extraction surgery and the improvement of implantation method, so as to avoid and reduce bacterial contamination and tumor cell dissemination in abdominal cavity.</p><p><b>METHODS</b>During June 2015 and May 2017, 48 cases of high located rectal or sigmoid colon cancer were operated laparoscopically with natural orifices specimen extraction surgery (NOSES) using a self-made specimen protecting sleeve. Operation indication: (1) Rectum and sigmoid colon cancer with the distance of more than 6 cm from tumor inferior margin to dentate line. (2) The maximum diameter of intestine together with mesangial and tumor <7 cm by intraoperative judgment. (3) No anal and distal rectal surgery, no anorectal stenosis or lack of expansion capacity caused by trauma. (4) No ulcerative colitis, Crohn's disease or radiation proctitis. After transecting the rectum, the specimen protective sleeve was inserted through the right lower 12 mm main Trocar (This sleeve was tailored from the laparoscopic protective sleeve produced by China 3L Corporation, which was intercepted with 25-35 cm from one end of the sleeve according to the length of distal rectal retention. One end was ligated and the other was open with a ligature band. About 5 ml paraffin oil was used to rinse and lubricate during the operation). The rectal stump retained 7-8 cm in abdominal cavity. The transanal ligation part of the protective sleeve was cut off, then the stapler nail seat was inserted and specimen was pull out through the sleeve and rectum.</p><p><b>RESULTS</b>There were 30 males and 18 females. The average age was (64.5±14.1) years, the BMI was (25.4±3.9) kg/m, the tumor diameter was (3.3±1.1) cm, the maximum diameter of specimen was (5.4±1.5) cm and the length of specimen was (18.6±4.3) cm. Among these 48 cases, specimens of 36 patients were pulled out through inside of the sleeve easily, while specimens of 12 patients were quite difficult with resistance. Of 12 cases, 7 needed the help of transverse forceps, 4 needed to make 1 cm incision in pull-through bowel and insert a suction to decrease the volume of large specimens with gathering of gas and fluid, and 1 received small abdominal incision to remove specimen and perform intestinal reconstruction due to big specimen (the diameter of tumor and mesentery was 7.5 cm). Specimen tears of 6 patients didn't result in dissemination thanks to the specimen protecting sleeve. The operation time was (113.2±76.1) min, the bleeding amount was (38.5±17.3) ml, the time to first oral intake was (47.9±4.4) h, and the postoperative hospitalization length was (8.5±1.7) d. Anastomotic leakage occurred in 1 case (2.1%). No intra-abdominal and trocar infection, and obstruction were found.</p><p><b>CONCLUSION</b>The use of protective sleeve and the improvement of the method of intraperitoneal implantation can effectively reduce the abdominal contamination during the specimen extraction. It can be applied to big specimens as well.</p>
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<p><b>OBJECTIVE</b>To identify a novel hemoglobinopathy applied by direct sequencing and clone sequencing.</p><p><b>METHODS</b>EDTA anticoagulated blood of proband and his parents were analyzed by hematology analyzers and Capillarys hemoglobin electrophoresis (CE). Then thalassemia genetypes were screened by gap-PCR and reverse dot blot (RDB). Proband was suspected with abnormal hemoglobin combine alpha beta compound thalassemia. The mutation of beta-globin was identified by direct sequencing and clone sequencing.</p><p><b>RESULTS</b>Hb analysis showed that probands Hb A2 variant was eluted in Z (C) zone and his father's in Z (A2) zone on CE,and proband's mother elevated HbA2 of 4.6%. Screened by RDB, the proband was CD71-72(+A) homozygote and showed the mismatch with his parents. Through direct sequencing and clone sequencing, we deduced that our proband inherited the mutations of HBB c.[219T>A;220G>T] from his father and inherited the Southeast-Asian deletion and HBB c.216-217insA from his mother.</p><p><b>CONCLUSION</b>A novel double heterozygote of HBB c.[219T>A; 220G>T] was identified in south China. This mutation enriches the beta-thalassemia gene mutation spectrum in Chinese population.</p>
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Child, Preschool , Humans , Male , Asian People , Genetics , Hemoglobins , Genetics , Hemoglobins, Abnormal , Genetics , Heterozygote , Mutation , Genetics , Pedigree , Thalassemia , Genetics , beta-Globins , GeneticsABSTRACT
In this research, we collected information of eighty nine sampling points of Isatidis Radix nationwide through data query and field survey, and the medicinal component contents of samples were determined by HPLC. By using Maxent Model and ArcGIS, along with ecological factor data, the national habitat suitability distribution of Isatidis Radix was predicted. R-language was adopted to establish a model of the relationship between the medicinal component contents and ecological factors. The medicinal quality was divided by ArcGIS grid computing. The results indicated that the three main ecological factors affecting the distribution of Isatidis Radix were precipitation in the driest season, mean annual temperature and mean temperature in the wet season. The suitable cultivation region of Isatidis Radix is mainly distributed in the north of China, but the medicinal quality is quite different, Isatidis Radix in Xinjiang province has higher medicinal quality. This study provides a reference for rational selection of planting areas of Isatidis Radix.
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[Objective]To evaluate the terminal hair growth of different body positions among in pregnant women ,analyze the contribution of each area to the diagnosis to hirsutism ,to improve the mFG scoring system.[Method]Pregnant women aged 20~41 years,with normal range of total testosterone levels and non-hirsute were recruited at their prenatal care in Sun Yat-sen Memorial Hospital,Sun Yat-sen University. They were followed up before pregnancy,at the 5th and the 9th week,the 10th and the 14th week, the 15th and the 20th week,the 21th and the 24th week of gestation. Then 72 more cases of pregnant women were recruited and followed up at 15~24th week. At each time of followed up,their total testosterone(TT)levels was examined by liquid chromatography tandem mass spectrometry(LC/MS-MS)and terminal hair growth were assessed by mFG score. Significant difference procedure least(LSD) analysis of variance was used to compare the levels of testosterone and mFG score in different gestational weeks. Receiver operating characteristics(ROC)analysis and logistics analysis were conducted to evaluate the contributory strength of hair growth in each body position for the diagnosis of hirsutism. The scores in the body area which made a significant contribution to the total were summed up and termed the simplified mFG score(sFG score). Following,the sFG scores were subjected to ROC analysis to determine the thresh-old that would maximize both the sensitivity and specificity of the measure to accurately distinguish hirsute from non-hirsute patients.[Results]Among the forty three pregnant women who were followed-up from before pregnancy to 24th week,the mean±SD for TT was (1.09 ± 0.59)nmol/L before pregnancy,and(1.13 ± 0.40),(1.28 ± 0.38),(1.83 ± 0.63),(1.82 ± 0.52)nmol/L for 5~ 9th,10~14th,15~20th week,and the mFG score was 1.65 ± 0.60,2.30 ± 0.45,3.60 ± 0.68,4.20 ± 0.41 and 4.40 ± 0.77,respectively. The order of the facial and body sites presented with new terminal hair growth was upper abdomen,lower abdomen,lower back,up-per lip,thighs,upper back,chest,upper arm,and chin,in sequence. After analyzing 115 cases(including the 72 cases recruited later),ROC analysis showed that the diagnostic value of different sites for hirsutism(mFG≥5):upper lip>lower back>thighs>lower abdomen>upper arm>upper back>chest/upper abdomen. Four sites among them ,namely upper lip ,lower back ,thighs and lower abdomen had the best diagnostic value,and the AUC for ROC were 0.779,0.728,0.675,and 0.626,respectively. Both ROC and logistic analysis indicated that he most significant body areas in defining hirsutism(defined as an mFG score≥5)were the upper lip, lower back,thighs,and lower abdomen. Using a cut-off value of 3,the combination of the four areas has the best sensitivity and specificity in distinguishing hirsute from non-hirsute women. [Conclusion]The study suggested that the mFG score increased as pregnancy progressed before the 24 weeks of gestation. The subset of upper lip,lower back,thighs and lower abdomen may be a reli-able simplification of mFG system for the evaluation of excess hair growth. The cut-off value was of≥3.
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OBJECTIVE:To establish the technical system that is suitable for protein extracting in Angelicae sinensis seed and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE),and provide technical support for detecting the protein quality and variety purity. METHODS:Using protein content and number of electrophoretic bands as indexes,8 methods,includ-ing voncentrated gel method,salt-soluble protein method,electrode buffer method,dimercaptosyl alcohol (DTT) method,urea method,mercaptoethanol method,trimethylolaminomethane (Tris) method,and acetone precipitation method,were conducted to extract the protein in A. sinensis seed and screen the optimal extraction method. Then based on optimal extraction method,effects of different materials-lipid ratios,sample dilution times(sample volume)and separate gel concentration on SDS-PAGE were investi-gated. RESULTS:Mercaptoethanol method extracted the highest protein contents(29.931 mg/g),with many electrophoretic bands and clear background. When mercaptoethanol method was used as optimal extraction method,electrophoresis effects were the best in the conditions of materials-lipid ratio of 1:10,sample volume of 5 times,separate gel concentration of 15%,which obtained 18 bands totally. CONCLUSIONS:Established protein extraction method and SDS-PAGE technical system are suitable for detecting the purity of A. sinensis seed.
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Objective To evaluate the quality of Isatidis Radix from different ecotypes by using projection pursuit model. Methods Totally 11 batches of Isatidis Radix from different ecotypes in Gansu Province were used as evaluation samples. With the contents of epigoitrin, uridine, guanosine, adenosine, benzoic acid, salicylic acid, indigo, indirubin, and alcohol extract as evaluating indexes, combined with projection pursuit model established by DPS V 9.50 statistics software, the quality of Isatidis Radix was evaluated. Results Based on the evalution of nine main indexes, quality order of the 11 batches of Isatidis Radix was acquired: S2>S1>S9>S7>S8>S3>S5>S10>S11>S4>S6. Conclusion The projection pursuit model was available for the quality evaluation of different ecotype Isatidis Radix. The quality of Isatidis Radix from different ecotypes is different significantly, and the quality of tetraploid Isatidis Radix of Shijiazhuang is the best.
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There is increasing evidence that microcirculation hypoxia plays an important role in pathogenesis of inflammatory bowel disease (IBD).Hypoxia-inducible factors (HIFs)are transcriptional factors that serve as master regulators in ischemic and hypoxia injuries.Aims:To investigate the effect of HIF-2αon dextran sulfate sodium (DSS)-induced colitis in mice and its possible mechanism.Methods:Mx-Cre/LoxP recombination system was utilized to establish a conditional HIF-2αgene knockout (HIF-2α-/-)mouse model.C57BL/6,HIF-2α+/+and HIF-2α-/-mice were randomly allocated into DSS colitis group and water drinking group,respectively.Experimental colitis was induced by treatment with 4% DSS in drinking water for 7 days,and the disease activity index (DAI)was assessed daily.Mice in each group were sacrificed on day 1,3,5 and 7 in batch;the histopathological changes of colonic tissue were observed, and mRNA expressions of HIF-2αand tumor necrosis factor-α(TNF-α)were measured by real-time PCR.Results:During model establishment,expression of HIF-2αmRNA in colonic tissue was elevated in C57BL/6 and HIF-2α+/+DSS colitis groups,and the DAI and colonic inflammatory score were significantly higher than those in C57BL/6 water drinking group (P<0.05 on day 5 and day 7).Compared with HIF-2α+/+DSS colitis group,HIF-2α-/-DSS colitis group had more severe colonic inflammatory injury and the DAI and inflammatory score were further increased (P all<0.05,except the inflammatory score on day 7);expression of TNF-αmRNA in colonic tissue was also increased significantly in HIF-2α-/-DSS colitis group (P<0.05 on day 5 and day 7).Conclusions:HIF-2αmay ameliorate colonic inflammatory injury in mice with DSS colitis via inhibition of TNF-αexpression.
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The study is aimed to research the relationship between the seedling grade of Codonopsis pilosula and yield and quality of medicinal materials, so as to provide basis for establishing seedling standard. Thirty seedlings of C. pilosula were collected from the main production areas in Gansu province, such as Weiyuan, Minxian, Zhangxian, Dangchang and Longxi, root length and diameter and weight of all the samples were measured. According to the clustering results, seedlings were divided into 3 levels, and field experiments were conducted with three levels seedling, yield and quality were tested in laboratory. Results have showed that emergence of grades 1 was faster than that of grades 2 and 3. Yield of grades 1 was significantly higher than that of grades 2 and 3 (P<0.05). Propargyl glycoside content of grades 1 was the highest, and significantly higher than that of grades 3. Polysaccharide content of grades 3 was the highest and significantly higher than that of grades 1 and 2 (P<0.05). So considering yield, quality and investment cost of C. pilosula, planting seedlings of C. pilosula should select that root length>15.6 cm, root diameter>2.7 mm, root weight>0.56 g.
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OBJECTIVE:To select the optimum conditions for germination test of Sophora alopecuroides seed,and to provide reference for formulating seed test rule and standardization of S. alopecuroides. METHODS:S. alopecuroides seeds were soaked in 98% H2SO4 for 30 min and 35 ℃ warm water for 29 h,and then treated under different temperature conditions (15 ℃,20 ℃, 25℃and 30℃),different germination beds(on paper,between paper,on sand,in sand)and different light conditions(2 000 lx lighting 16 h,dark). Optimal germination condition was screened by using germination rate,germinative potential and germinative index as indicators. S. alopecuroides seeds were cultured under this condition for 7 days,and then germination rate was determined. RESULTS:Different temperatures had no significant effect on germination rate,but influenced germinative potential and germina-tive index;those indicators reached maximal value at 20 ℃. Different germination beds affected each indicator,and under condi-tion of on sand,those indicators were the highest. Light treatment had no significant effect on germination indicators. Under suit-able condition,the seed sprouted since first day of germination bed treatment;germination rate was more than 90% on second day,and reached maximal value on fifth day and didn’t increase any longer. CONCLUSIONS:The suitable condition of seed ger-mination was soaking in 98% H2SO4 30 min+35 ℃ warm water for 29 h,on sand under light at 20 ℃. Initial count on second day of germination bed treatment and final count on forth day were analyzed statistically as well as germination rate. This method can be used as standard quality test of the seed of S. alopecuroides.
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OBJECTIVE:To select the optimum conditions for germination test of Astragalus membranaceus seed and provide reference for the formulation of testing rules of A. membranaceus seed. METHODS:First,the dormancy of A. membranaceus seeds were broken by soaking seed with 98% H2SO4 for 30 min and 35 ℃ warm water for 9 h,then treated with different temperatures of germination,different germinating beds and light conditions. Different treatment methods were evaluated by germination rate, germination potential and germination index. RESULTS:With the increase of temperature,germination rate,germination potential and germination index decreased. At 30 ℃,germination rate,germination index,and germination potential were significantly low-er than those of seed with other treatments,in which the germination rate was the highest at 15℃;but there wasn’t significant dif-ference in germination rate of 15 ℃ and 20 ℃. There were no significant differences in germination rate of different germination beds,but sand bed could restrain bacterial growth well. Under both light and dark conditions,seed could normally germinate. The appropriate condition of A. membranacus seed germination was at 15 ℃,sand bed and dark. The germination rate,germination po-tential and germination index were(98.5±0.65)%,(85.5±0.87)%and 175.8±2.31,respectively. Meanwhile,the germination pe-riod was only 4 days. CONCLUSIONS:Suggested quality control method of A. membranaceus seed is that at 15 ℃,sand bed and dark. The second day of germination as initial counting time and forth day as the last counting time are used to calculate germina-tion index. This method is easy and controllable. It also has short germination period and high germination rate.
ABSTRACT
Objective To explore the therapeutic effect and safety of high-frequency diathermic therapy combined with oral S-1 and intraperitoneal perfusion chemotherapy on malignant seroperitoneum of advanced gastric carcinoma.Methods Fifty-two advanced gastric carcinoma patients with malignant seroperitoneum were divided into observation group and control group.Twenty-five patients in observation group were received DDP 60 mg/m2 combined with 5-Fu 600 mg/m2 and IL-2 3 000 000 U intraperitoneal perfusion chemotherapy,as well as oral S-1 of 40mg bid on day 1-14 of every 21 days for a total of two or three cycles.High-frequency diathermic therapy was administered thirty minutes after intraperitoneal perfusion chemotherapy,twice a week for 3 weeks.Twenty-seven patients in control group were treated with S-1 and intraperitoneal perfusion chemotherapy only.Results In observation group,4 patients were CR,15 patients were PR,5 patients were SD,1 patient was PD,and the response rate (CR+PR) was 76 ％.In control group,3 patients were CR,14 patients were PR,7 patients were SD,3 patients were PD,and the response rate (CR+ PR) was 63 ％ (P ＞ 0.05).Median survial time for observation group was 10.0 months,8.0 months for control group (P ＞ 0.05).Karnofsky scores after treatment in observation group was higher than that in control group,patients' clinical benefit rate were 80.0 ％ (20/25) and 51.9 ％ (14/27).The difference in life quality between the two groups was statistically significant (P ＜ 0.05).There was no significant difference in adverse reactions between the two groups (x2 =4.544,P =0.033).Conclusion High-frequency diathermic therapy combined with oral S-1 and intraperitoneal perfusion chemotherapy for advanced gastric carcinoma with malignant seroperitoneum has good short-term curative effects and security,and this method should be further studied.