Bushen Huoxue Fang improves recurrent miscarriage in mice by down-regulating the JAK2/STAT3 pathway / 南方医科大学学报

OBJECTIVE@#To investigate the efficacy of Bushen Huoxue Fang (BSHXF, a traditional Chinese medicine formula) for improving recurrent spontaneous abortion (RSA) in mice and the role of tyrosine kinase (JAK2) and transcriptional activator (STAT3) signaling pathway in its therapeutic mechanism.@*METHODS@#Female CBA/J mice were caged with male DBA/2 mice to establish RSA mouse models, which were randomly divided into model group, dydrogesterone group and BSHXF group, with the female mice caged with male BALB/c mice as the control group (n=6). From the first day of pregnancy, the mice were subjected to daily intragastric administration of BSHXF, dydrogesterone, or distilled water (in control and model groups) for 12 days. After the treatments, serum levels of antithrombin III (AT-III), activated protein C (APC), tissue plasminogen activator (t-PA), progesterone, human chorionic gonadotropin (HCG), and estradiol (E2) were detected in each group using ELISA. HE staining was used to observe the morphological changes of the endometrium of the mice. Western blotting was performed to determine the expressions of p-JAK2, p-Stat3 and Bcl-2 in the placenta of the mice.@*RESULTS@#Compared with the control mice, the mouse models of RSA showed a significantly increased embryo loss rate with decreased serum levels of AT-III, T-PA, progesterone, APC and HCG, increased placental expressions of p-JAK2, p-STAT3 and Bax, and decreased expression of Bcl-2 (P < 0.05). Treatments with BSHXF and dydrogesterone both increased serum levels of AT-III, t-PA and HCG in the mouse models; Serum APC level was significantly reduced in BSHXF group and serum progesterone level was significantly increased in dydrogesterone group (P < 0.05).@*CONCLUSION@#BSHXF can improve the prethrombotic state and inhibit cell apoptosis by downregulating the JAK2/STAT3 pathway to increase the pregnancy rate in mouse models of RSA.

Effect of heavy metal lead on anticoagulant activity of leech based on label-free quantitative proteomics / 中国中药杂志

Whitmania pigra is the most widely distributed species of leeches in the market. In this study, the effect of heavy metal lead pollution on the anticoagulant activity of Wh. pigra was studied and the potential mechanism was explored. Pb(NO_3)_2 was used to contaminate the breeding soil which was then used to rear Wh. pigra for 50 days(lead-contaminated group, LC group), and meanwhile the blank control group(CG group) was set. Proteins were extracted from the obtained leech samples, and the differentially expressed proteins between LC and CG groups were analyzed by label-free proteomics technology. In this study, a total of 152 differentially expressed proteins were screened out, of which 93 proteins were up-regulated and 59 proteins were down-regulated in LC group. Bioinformatics analysis showed that the biological processes enriched with the differentially expressed proteins were mainly vesicle-mediated transport and transport positive regulation; the enriched cell components were mainly endocytosis vesicles and apical plasma membrane; the enriched molecular functions mainly included carbohydrate binding. The differentially expressed proteins were enriched in 76 KEGG pathways, which mainly involved metabolic pathways, biosynthesis of secondary metabolites, and bacterial invasion of epithelial cells. In this study, two differentially expressed proteins with Antistasin domain were presumed, which provides reference for further exploring the regulatory mechanism and signal transduction underlying the effect of lead pollution on the anticoagulant activity of leech.

Determination of caffeine content in Ginkgo Folium by ultra high performance liquid chromatography-triple quadrupole mass spectrometry / 中国中药杂志

In this experiment, an ultra-high performance liquid chromatographytandem triple quadrupole mass spectrometry was established for the determination of caffeine in commercially available Ginkgo Folium. The samples were extracted by ultrasonic method with methanol, and separated on Waters CORTECS T3 column(2.1 mm×100 mm, 2.7 μm), with mobile phase of 0.1% formic acid solution-0.1% formic acid acetonitrile solution for gradient elution, at flow rate of 0.3 mL·min~(-1); column temperature of 30 ℃, and injection volume of 2 μL. Mass spectrometry was conducted at ESI~+ multiple reaction monitoring(MRM) mode; quantitative analysis was conducted with external standard method. The results showed that in the range of 0.099 6-9.96 ng·mL~(-1), there was a good linear relationship between the mass concentration of caffeine and the peak area, R~2=0.999; the average recovery was 84.51%, with RSD of 6.2%. The results of precision, repeatability and stability showed that the RSD was 5.1%, 5.9%, 7.2%, respectively. The content range of caffeine in 10 batches of Ginkgo Folium was 1.52-60.86 μg·kg~(-1). In conclusion, this method is accurate, reliable and reproducible, which provides a reference for the safety study of Ginkgo Folium.

Prognostic Value of TP53 Mutation for Transcatheter Arterial Chemoembolization Failure/Refractoriness in HBV-Related Advanced Hepatocellular Carcinoma / Journal of the Korean Cancer Association, 대한암학회지

Purpose@#This study aimed to investigate the clinicopathologic features and mutational landscape of patients with hepatitis B virus (HBV)–related advanced hepatocellular carcinomas (HCC) undergoing transcatheter arterial chemoembolization (TACE). @*Materials and Methods@#From January 2017 to December 2018, 38 patients newly diagnosed with HBV-related advanced HCC were enrolled in the final analysis. Their pathological tissues and corresponding blood samples before TACE treatment were collected for whole-exome sequencing. Response to TACE was evaluated at 1-3 months after two consecutive use of TACE. Predictive factors were analyzed by univariate and multivariate analyses in a bivariate Logistic regression model. Enrichment of related pathways of all driver genes were acquired using the gene set enrichment analysis (GSEA). @*Results@#Among 38 patients, 23 (60.5%) exhibited TACE failure/refractoriness. Patients with TACE failure/refractoriness showed higher frequency of TP53 mutation than their counterparts (p=0.020). Univariate and multivariate analyses showed that only vascular invasion and TP53 mutation were significantly correlated with TACE failure/refractoriness in HBV-related advanced HCC. Of the 16 patients without vascular invasion, eight (50.0%) had TP53 mutations, and TP53 mutation was associated with TACE failure/refractoriness (p=0.041). Moreover, GSEA showed that mitogen-activated protein kinase and apoptosis pathways induced by TP53 mutation were possibly associated with TACE failure/refractoriness. @*Conclusion@#Our study suggested that TP53 mutation was independently related with TACE efficacy, which may work via mitogen-activated protein kinase and apoptosis pathways. These findings may provide evidence to help distinguish patients who will particularly benefit from TACE from those who require more personalized therapeutic regimens and rigorous surveillance in HBV-related advanced HCC.

Identification of Decoction Pieces and Standard Decoction of Ophiopogonis Radix and Liriopes Radix by TLC / 中国实验方剂学杂志

Objective:There were 92 kinds of compound preparations containing Ophiopogonis Radix in the 2015 edition of Chinese Pharmacopoeia， but there was no effective method to identify these compound preparations. Because Ophiopogonis Radix and Liriopes Radix are similar in appearance， it is easy to be confused in application. The aim of this study was to set up a thin layer chromatography （TLC） to identify compound preparations containing Ophiopogonis Radix and distinguish Ophiopogonis Radix and Liriopes Radix in the forms of decoction pieces and standard decoction. Method:In this study， decoction pieces of Ophiopogonis Radix and Liriopes Radix were collected and separately prepared as standard decoction. TLC was used to qualitatively identify decoction pieces and standard decoction of Ophiopogonis Radix and Liriopes Radix， and compound preparations containing Ophiopogonis Radix. In the TLC， the lower solution of chloroform-methanol-water （65∶35∶10） was selected as the developing agent and 10% sulfuric acid ethanol solution as the chromogenic agent. Result:The resolution of this TLC was good. Decoction pieces， standard decoction and preparations of Ophiopogonis Radix had the same characteristic strips， which were two bright white fluorescent strips under ultraviolet lamp （365 nm）. But these two characteristic strips were not existed in the TLC of decoction pieces and standard decoction of Liriopes Radix. The corresponding components of both of these two strips were identified as mixture containing saponins by LC-MSn， including ophiopogonin Ra， Tb， ophiopogonin D'， borneol glycoside， ophiopogonin C and Liriope muscari baily saponins C. Conclusion:The established TLC method， which has significant advantages such as high specificity and sensitivity， can be applied to the characteristic identification of decoction pieces and standard decoction of Ophiopogonis Radix， the identification of compound preparations containing Ophiopogonis Radix， and the distinction of Ophiopogonis Radix and Liriopes Radix， thus serving as an effective method to qualitatively identify Ophiopogonis Radix and its compound preparations.

Relationship Between Cardiac Troponin Ⅰ Autoantibody and Left Ventricular Remodeling in Patients With Acute Myocardial Infarction / 中国循环杂志

Objectives: To analyze the relationship between cardiac troponin I autoantibody (cTnIAAb) and left ventricular remodeling in patients with acute myocardial infarction (AMI). Methods: A total of 131 AMI patients were enrolled. Serum levels of cTnIAAb were measured by ELISA. Echocardiography was examined at the onset of AMI and 1 year follow-up evaluation. Taking left ventricular end systolic volume (LVESV) increasing>15% as left ventricular remodeling, 2-classified logistic stepwise regression analysis was conducted to screen 12 risk factors related to left ventricular remodeling. Results: 23/131(17.6%) patients were with positive cTnIAAb and 82.4% with negative cTnIAAb. 49 patients lost contact and in the rest 91 patients, 21.1% were with positive cTnIAAb. Clinical information was similar between cTnIAAb positive and negative patients upon admission, P>0.05; echocardiography showed that 28 (42.2%) patients had LVESV increasing>15% by 1 year follow-up study whom including 10 (52.6%) patients with cTnIAAb positive and 18 (25.4%) negative. 2-classified logistic stepwise regression analysis indicated that BNP peak and positive cTnIAAb were the risk factors for left ventricular remodeling (OR=1.001, 95% CI 1.001-1.002) and (OR=3.552, 95% CI 1.148-10.989), both P=0.028. Conclusions: Serum cTnIAAb was positive in part of AMI patients which was related to increased risk of left ventricular remodeling; cTnIAAb might be involved in pathophysiological process of left ventricular remodeling in AMI patients.

Expert consensus on prescription comment of Chinese traditional patent medicine for promoting the rational use of drugs in Beijing / 中国中药杂志

With the growth of number of Chinese patent medicines and clinical use, the rational use of Chinese medicine is becoming more and more serious. Due to the complexity of Chinese medicine theory and the uncertainty of clinical application, the prescription review of Chinese patent medicine always relied on experience in their respective, leading to the uncontrolled of clinical rational use. According to the traditional Chinese medicine (TCM) theory and characteristics of the unique clinical therapeutics, based on the practice experience and expertise comments, our paper formed the expert consensus on the prescription review of Chinese traditional patent medicine for promoting the rational use of drugs in Beijing. The objective, methods and key points of prescription review of Chinese patent medicine, were included in this expert consensus, in order to regulate the behavior of prescription and promote rational drug use.

Standardization of names in prescriptions of traditional Chinese medicines / 中国中药杂志

Chinese medicine prescriptions are a type of medical documents written by doctors after they understand the patients' conditions for syndrome differentiation. Chinese medicine prescriptions are also the basis for pharmacy personnel to dispense medicines and guide patients to use drugs. It has the legal, technical and economic significances. Chinese medicine prescriptions contain such information of names, quantity and usage. Whether the names of drugs in Chinese medicine prescriptions are standardized or not is directly related to the safety and efficacy of the drugs. At present, nonstandard clinical prescriptions are frequently seen. With "Chinese medicine prescription", "names of drug in Chinese medicine prescription" and "standards of Chinese medicine prescription" as key words, the author searched CNKI, Wanfang and other databases, and consulted nearly 100 literatures, so as to summarize current names of drugs in traditional Chinese medicine prescription, analyze the reasons, and give suggestions, in the expectation of standardizing the names of drugs used in traditional Chinese medicine prescriptions.

AFT024-SCF construction and functional identification / 中国病理生理杂志

AIM:To construct AFT024-SCF cell line and HPC-Lhx2 cell line for confirming the biological function of AFT024-SCF.METHODS:The HPC-Lhx2 cell line, AFT024-SCF cell line and AFT024-GFP cell line were constructed by retro-viral infection.The expression of stem cell factor(SCF) in AFT024-SCF cells was detected by real-time PCR and Western blot.SCF in the supernatant of AFT024-SCF was detected with ELISA.The supernatant of AFT024-SCF and AFT024-GFP were collected and then diluted (1:10) with basic IMDM medium.So we made 4 culture medium:AFT024-SCF medium was used for experiment group, AFT024-GFP medium was used for endogenous negative control, IM-DM basic medium was used for exogenous negative control, and IMDM basic medium with SCF was used for positive con-trol.SCF-dependent HPC-Lhx2 cell line was cultured in these 4 different medium for 72 h.According to MTT method and colony forming experiment, the biological function of AFT024-SCF was confirmed by the proliferation ability of SCF-depend-ent HPC-Lhx2 cell line.RESULTS:SCF was highly expressed in AFT024-SCF cells.After cultured for 72 h, neither IM-DM basic medium nor GFP-AFT024 medium support HPC-Lhx2 cell line proliferation.However, AFT024-SCF medium supported HPC-Lhx2 cell line expansion as well as the positive control medium.CONCLUSION:AFT024-SCF cells ex-press SCF successfully and recombinant SCF can be replaced by the supernatant of AFT024-SCF culture medium for expan-ding HPC-Lhx2 cell line in vitro.

Value of flowcytometry fluorescence immunoassay of CA242 in the diagnosis of pancreatic cancer / 中华胰腺病杂志

Objective To conduct a methodological evaluation on flowcytomotry fluorescence detection of serum CA242, and to evaluate its value in the diagnosis of pancreatic cancer.Methods The blood samples of 40 cases of pancreatic cancer, 49 cases of other tumors, 48 cases of benign digestive diseases and 159 healthy volunteers were collected.Fowcytometry fluorescence immunoassay was used to detect serum level of CA242, and it was compared with routine ELISA method to measure its sensitivity and specificity.Results The detection limit of CA242 by flowcytometry fluorescence immunoassay was 0.89 U/ml, the linear range was 1～ 500 U/ml after confirmation.The within-batch CV was 3.37％～ 5.30％, between batch CV was 7.43％ ～ 9.60％.When compared with routine ELISA, flowcytometry fluorescence immunoassay showed the equation of linear regression is Y =1.0398X-0.947, r =0.9687.Area under ROC curve was 0.811 ± 0.025 (95％ CI 0.763 ～0.859), with 18.625 U/ml as the best cutoff value, the specificity was 92.0％ and the sensitivity was 62.1％.Conclusions Flowcytometry fluorescence immunoassay for CA242 testing has the advantage of shorter detection time, miltiple sample and testing project detection, which is worth of clinical application.

Preparation process of apigenin particles by supercritical CO2 anti-solvent method / 中草药

Objective: To prepare the apigenin particles by supercritical CO2 anti-solvent technology, to optimize the preparation process of apigenin particles on the basis of single factor experiments using the average particle size as evaluation index through orthogonal test design, and to investigate the in vitro dissolution rate. Methods: The particle size distribution, scanning electro- microscope (SEM) analysis, infrared spectrum (IR), and differential scanning calorimetry (DSC) were used to validate the selected process. The external dissolution rates of apigenin ingredient and apigenin particles of the optimal process were tested to speculate the bioavailability of apigenin. Results: The optimal process conditions by orthogonal test were set as follows: The proportion of acetone and dimethyl sulfoxide (DMSO) was 24:1, temperature was 40°C, pressure was 10 MPa, solution volumetric flow rate was 2.0 mL/min, and mass concentration of apigenin was 8 mg/mL. Under the optimal conditions, the volume average particle size was obviously smaller than that of ingredient. And the shapes of apigenin were irregular. FTIR and DSC analyses showed that the chemical structure did not change. The results showed that the in vitro dissolution rate of apigenin particles of the optimal process was significantly larger than that of apigenin ingredient. Conclusion: The supercritical CO2 anti-solvent technology is feasible to prepare apigenin particles and promote its bioavailability, and it provides a reference basis for preparing ultrafine particles.

An improved association-mining research for exploring Chinese herbal property theory: based on data of the Shennong's Classic of Materia Medica / 中西医结合学报

Knowledge Discovery in Databases is gaining attention and raising new hopes for traditional Chinese medicine (TCM) researchers. It is a useful tool in understanding and deciphering TCM theories. Aiming for a better understanding of Chinese herbal property theory (CHPT), this paper performed an improved association rule learning to analyze semistructured text in the book entitled Shennong's Classic of Materia Medica. The text was firstly annotated and transformed to well-structured multidimensional data. Subsequently, an Apriori algorithm was employed for producing association rules after the sensitivity analysis of parameters. From the confirmed 120 resulting rules that described the intrinsic relationships between herbal property (qi, flavor and their combinations) and herbal efficacy, two novel fundamental principles underlying CHPT were acquired and further elucidated: (1) the many-to-one mapping of herbal efficacy to herbal property; (2) the nonrandom overlap between the related efficacy of qi and flavor. This work provided an innovative knowledge about CHPT, which would be helpful for its modern research.

Study on effect of Curculiginis Rhizoma and its active ingredient on PXR-CYP3A of L02 cells in different states / 中国中药杂志

<p><b>OBJECTIVE</b>To define the effect of Curculiginis Rhizoma and its active ingredient orcinol glucoside on PXR-CYP3A of L02 cells in normal and deficiency-cold states, in order to lay a foundation for studies on the mechanism of efficacy expression differentiation of Curculiginis Rhizoma in different states.</p><p><b>METHOD</b>Serums of normal and deficiency-cold rats were adopted to culture L02 cells and induce cells in normal and deficiency-cold states. After aqueous extracts from Curculiginis Rhizoma and its active ingredient orcinol glucoside were used in cells in different states, PXR protain expression and CYP3A activity of L02 cells in normal and deficiency-cold states were observed.</p><p><b>RESULT</b>MTT results showed that aqueous extracts from Curculiginis Rhizoma and orcinol glucoside could significantly enhance viability of L02 cells. Aqueous extracts from Curculiginis Rhizoma could significantly reduce PXR protein expression of L02 cells in normal state, while orcinol glucoside could significantly reduce CYP3A activity and PXR protein expression of L02 cells in normal state. Meanwhile, aqueous extracts from Curculiginis Rhizoma could significantly increase CYP3A activity and PXR protein expression of L02 cells in deficiency-cold state, while orcinol glucoside could significantly reduce CYP3A activity and increase PXR protein expression of L02 cells in deficiency-cold state.</p><p><b>CONCLUSION</b>Curculiginis Rhizoma can activate PXR and induce CYP3A activity of L02 cells in deficiency-cold state, but with no effect or even counteraction on PXR and its induced CYP3A of L02 cells in normal state.</p>

Inhibition effect of small interfering RNA targeting connective tissue growth factor on liver fibrosis in rats / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To design and synthesize small interfering RNA (siRNA) targeting connective tissue growth factor (CTGF) and to investigate its effect on liver fibrosis.</p><p><b>METHODS</b>The interference sequence of CTGF was designed and synthesized. Rat hepatic fibrosis model was induced by intraperitoneal injection of 40 % CCl4(3 ml/kg). Thirty male rats were randomly divided into 5 groups: in normal control and model groups rats received tail vein injection of normal saline every 3 days for 8 consecutive weeks; in preventive group rats received tail vein injection of CTGF siRNA (0.1 mg/kg) every 3 days for 8 weeks; in 2-w treatment group CTGF siRNA was given for 6 weeks starting from two weeks after CCl4 injection; in 4-w treatment group CTGF siRNA was given for 4 weeks starting 4 weeks after CCl4 injection. The serum and hepatic tissue samples were harvested 3 days after the last CCl4 injection. Hepatic fibrosis indices were measured. Expression of CTGF mRNA and protein in the liver was evaluated by RT-PCR and Western blot, respectively. Fibrosis in rat liver was analyzed by Masson staining.</p><p><b>RESULTS</b>Compared with model group (0.544 0.019), the expression of CTGF mRNA and protein in liver of both preventive(0.105 ± 0.003) and 2-w treatment groups (0.190 ± 0.006) were markedly down-regulated (P<0.05). Inflammation, necrosis and fibrosis in hepatic tissue were significantly attenuated. In addition, the serum ration of liver fibrosis indices was greatly reduced(P<0.05). Compared with preventive and 2-w treatment groups, the expression of CTGF mRNA and protein in liver in 4 weeks of treatment group were up-regulated (P<0.05); inflammation, necrosis and fibrosis in hepatic tissue were relative increased; and the serum concentrations of liver fibrosis indices were relatively higher (P<0.05).</p><p><b>CONCLUSION</b>The highly effective CTGF siRNA has been successfully synthesized, which can inhibit CTGF expression in liver, prevent hepatic fibrosis and its progress in rats.</p>

Extractible properties of PDLLA under different extraction conditions / 生物医学工程学杂志

This assay was made to investigate the extractible properties of PDLLA under different extraction conditions. The ratio of test sample to extraction medium was 0.2 g to 1 ml. The distilled water, artificial saliva, Eagle's MEM and hexane were selected respectively as the extraction medium. The samples were extracted under 37 degrees C 24 h, 72 h, 1 w, 2 w and 3 w; 50 degrees C 72 h; 70 degrees C 24 h and 72 h; 121 degrees C 1 h. By use of gas chromatography, the ethyl alcohol, xylene and ethyl acetate content were measured. The results showed the ethyl alcohol content < 1.998 ug/ml, xylene contents was < 53.39 ug/ml and ethyl acetate content < 3.647 ug/m of PDLLA in distilled water, artificial saliva and Eagle's MEM under the condition of 37 degrees C from 24 h to 3 w. The ethyl alcohol content and xylene content in hexane were higher than those in the other three aqueous solutions. When the extracted temperature was increased, the contents of above three components were kept at the original level. There was almost no difference in the extractible properties of PDLLA among distilled water, artificial saliva and Eagle's MEM. The results did not change even if the extraction time and temperature were increased. It is a new concept to evaluate the safety of biomaterials by combining chemical and biological extraction tests, which will be significance in narrowing the gap between physical-chemical tests and biological tests for medical devices.