To investigate the effect of recombinant human growth hormone (rhGH) on JAK2-STAT3 pathway and the growth of gastric cancer cell lines at different GHR expression status, the eukaryotic expression vector targeting human GHR (pGPU6/GFP/Neo-shGHR and pGPU6/GFP/Neo-scramble) was constructed and transfected into MGC803 cells by Lipofectamine 2000. Stable expressive cell lines were obtained by G418 screening. The expression of GHR was analyzed by Western blotting. After being stimulated with rhGH, cell growth was detected by MTT assay. Cell cycle and apoptosis were examined by flow cytometry. The components of JAK2/STAT3 signaling pathway were detected by Western blotting. There is no significant difference of GHR expression between MGC803 and pGPU6/GFP/Neo-scramble-transfected cells (named as MGC803-NC) (P > 0.05). Compared with MGC803, the GHR expression in pGPU6/GFP/Neo-shGHR-transfected cells (named as MGC803-shGHR) decreased significantly (protein decreased 50%). The cells were treated with rhGH at 0, 150 and 300 ng x mL(-1), the growth rate of MGC803 and MGC803-NC increased significantly, PI and the number of G2/M phase cells all increased significantly, and apoptosis decreased significantly. Western blotting revealed that the expression of pJAK2 and pSTAT3 was up-regulated after being treated with rhGH in MGC803 and MGC803-NC cells. In contrast, similar change was not observed in MGC803-shGHR cells. Knockdown of GHR gene may decrease the sensitivity of gastric cancer cells to rhGH, and down-regulating of components of the expression of JAK2/STAT3 signaling pathway may be the potential mechanisms.
Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Human Growth Hormone ; genetics ; pharmacology ; Humans ; Janus Kinase 2 ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, Somatotropin ; genetics ; metabolism ; Recombinant Proteins ; genetics ; pharmacology ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Stomach Neoplasms ; metabolism ; pathology ; Transfection

OBJECTIVETo investigate the chemical constituents of effective part of Epimedium brevicornum.

METHODThe sample was extracted with ethanol and purified by macroporous resin. The structures were identified by HPLC-MS3 experiments.

RESULTNine compounds were identified from the effective part of E. brevicornum.

CONCLUSIONThe method is simple and rapid for the identification of the flavonoids from E. brevicornum.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Epimedium ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Reproducibility of Results

OBJECTIVETo explore the prediction of the site for microsurgical vasoepididymostomy (VE) in the treatment of epididymal obstructive azoospermia (OA).

METHODSThis study involved 56 infertile men with confirmed OA whose obstruction was suspected to be in the epididymis. Based on their medical history and results of preoperative physical examination and ultrasonography, we predicted the sites for VE. We performed surgical scrotal exploration for the status of epididymal obstruction, conducted palpation and microscopic observation for the epididymal tubules to be anastomosed, and finally decided on the sites for VE by making sure of the presence of motile sperm in the epididymal fluid of the patients. After surgery, we followed up the patients for the rate of pregnancy.

RESULTSAll the patients received bilateral scrotal ultrasonography and surgical scrotal exploration, totaling 112 procedures, including 98 VE procedures. The accuracy rate of the predicted sites for VE was 80.5% (153/190) by medical history and physical examination, 80.3% (90/112) based on the results of ultrasonography, and 87.4% (90/103) according to the first selected epididymal tubules. Of the 28 patients followed up for more than 12 months, motile sperm were found in 19 (67.9% ) at 2 to 12 months and spontaneous pregnancies were achieved in 10 (35.7%), all with the anastomotic sites in the corpus or cauda.

CONCLUSIONMedical history and physical examination contribute to the selection of anastomotic sites and non-invasive scrotal ultrasonography is effective and practical for positioning epididymal obstruction. The epididymal tubules with motile sperm for anastomosis could be easily obtained from the most dilated ones in indurated epididymides.

Azoospermia ; surgery ; Body Fluids ; Epididymis ; diagnostic imaging ; surgery ; Female ; Humans ; Male ; Microsurgery ; methods ; Pregnancy ; Pregnancy Rate ; Scrotum ; diagnostic imaging ; Ultrasonography ; Vas Deferens ; diagnostic imaging ; surgery

In order to study the development characteristics of Rehmannia glutinosa tuberous root expansion and reveal the regulation mechanism of the genes related to hormones in this process, R. glutinosa "wen-85" was used as the experimental material in this study. R. glutinosa tuberous roots of different developmental stages were collected to observe phenotype and tissue morphology using resin semi-thin sections method. The genes related to hormone biosynthesis and response were chosen from the transcriptome of R. glutinosa, which was previously constructed by our laboratory, their expression levels at different development stages were measured by real-time quantitative PCR. The results showed that the root development could be divided into six stages: seeding, elongation, pre-expanding, mid-expanding, late-expanding and maturity stage. The anatomic characteristics indicated that the fission of secondary cambium initiated the tuberous root expansion, and the continuous and rapid division of secondary cambium and accessory cambium kept the sustained and rapid expansion of tuberous root. In addition, a large number oleoplasts were observed in root on the semi-thin and ultra-thin section. The quantitative analysis suggested that the genes related to biosynthesis and response of the IAA, CK, ABA,ethylene, JA and EB were up-regulated expressed, meanwhile, GA synthesis and response genes were down-regulated expressed and the genes of GA negative regulation factors were up-regulated expressed. The maximum levels of most genes expression occurred in the elongation and pre-expansion stage, indicating these two stages were the key periods to the formation and development of tuberous roots. Oleoplasts might be the essential cytological basis for the formation and storage of the unique medicinal components in R. glutinosa. The results of the study are helpful for explanation of development and the molecular regulation mechanism of the tuberous root in R. glutinosa.
Gene Expression Regulation, Developmental ; drug effects ; genetics ; Gene Expression Regulation, Plant ; drug effects ; genetics ; Lipid Droplets ; metabolism ; ultrastructure ; Microscopy, Electron, Transmission ; Plant Growth Regulators ; biosynthesis ; pharmacology ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; genetics ; growth & development ; metabolism ; Rehmannia ; genetics ; growth & development ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors

Objective To evaluate the reliability, validity and sensitivity of a Family Burden Scale (FBS) of disease used on schistosomiasis. Methods 224 schistosomiasis patients were investigated, using the FBS. Reliability was estimated by Cronbach's α coefficient and split-half reliability. Validity was tested by factor analysis. Sensitivity was evaluated by comparison of patients with different income levels. Results The Cronbach's α coefficient was 0.874 and split-half reliability was 0.939 for FBS, respectively. Most values of Cronbach's α and split-half reliability for each component of scale were above 0.70. Construct validity was appraised by factor analysis, and 6 factors were identified. These factors could explain 66.76 % of the total variance. Patients with different income levels showed significant difference in terms of family burden for schistosomiasis (P<0.001 ). Conclusion This FBS appeared to have satisfactory reliability, validity and sensitivity and could be used in evaluating family burden of schistosomiasis patients.

Aim To explore the optimal way of breeding and genotype identification of Arrb2 knockout mice, and to find a simple and quick polymerase chain reaction ( PCR) method for the genotyping of Arrb2 knockout mice. Methods Breeding homozygote genotype of Arrb2 gene knockout mice were copula-ted with wild-type C57BL/6J mice, and then the heterozygous mating were used for mating. The growth and development of off-spring were observed. The genomic DNA was extracted from the tail of two-week-old mice. PCR was employed to amplify the Arrb2 gene fragment, and electrophoresis was used to present the gene type. Results The breeding and reproducing were successful and three genotype offspring, including wild-type,heterozygous and homozygous knockout mice were obtained. Agarose gel electrophoresis results showed the size of PCR prod-ucts was about 186 bp and 224 bp, which was consistent with the expected target gene fragment, and identified Arrb2 gene knockout mice of different genotypes successfully. Western blot analysis demonstrated the lack ofβ-arrestin2 protein in the major organs from Arrb2 -/ - mice compared with Arrb2 +/ + and Arrb2 +/ - mice. Conclusions It is feasible to obtain the homo-zygous Arrb2 knockout mice by inbreeding heterozygotes. It is simple, rapid and reliable to identify mouse genetype by PCR.

OBJECTIVE: To develop the visual uroflow scale (VUS), analyze the relationship of VUS score and index of free uroflowmetry, assess urination function preliminarily and improve the work efficiency in the clinic. METHODS: Male lower urinary tract symptoms (LUTS) patients, who attended the Department of Urology in Peking University People's Hospital from March 2016 to March 2017, were assessed for their urination function according to the Visual Uroflow Scale without help from clinicians before undertaking a free uroflowmetry test. And afterwards, a free uroflowmetry was undertaken, and variables including maximal flow rate (Qmax), the average flow rate (Qave) and voiding volume (VV) was obtained. During the study, 124 cases were collected and 53 cases met the inclusion and exclusion criteria and were included in the study cohort. The Spearman correlation analysis was used for analyzing the correlation of VUS scores with free uroflowmetry variables and age. The validity of VUS was evaluated. RESULTS: Most of the patients could choose the very figure matched with self-condition by first instinct without any help from the clinician. The data were analyzed by Spearman correlation analysis. In the present study, voiding time was positively correlated with the VUS score (correlation coefficient, 0.62, P < 0.05). In the present cohort, the patients chose the third and fourth figures to take longer time to urinate, implying worse LUTS situation. Flow time and VUS scores were positively correlated (correlation coefficient, 0.61, P < 0.05). The patients with higher VUS scores would spend more time on urinate, no matter how long urinary hesitation was. Both Qmax and Qave were negatively correlated with the VUS score (correlation coefficient －0.54, －0.62, P < 0.05). The study illustrated that the VUS score suggested that the Qmax basically and further reflected the urination function. And its relationship to age revealed the decreased urination function of aging male, which had reached a consensus. CONCLUSION Development of VUS has helped the clinician assess the urination function preliminarily at the first time. Patients are assessed for a VUS score before getting surgery or receiving the drug for treatment, and can be re-assessed after. The VUS score can provide an objective quantitative basis to evaluate the treatment efficacy. In addition, considering that it is convenient, timesaving and easy to understand, the VUS is available for follow-up.
Cohort Studies ; Humans ; Lower Urinary Tract Symptoms ; Male ; Urination ; Urodynamics

Objective To study the effect of intestinal flora on the pharmacokinetics of ticagrelor in rats.Methods A total of 45 SD rats were randomly divided into control group,test A group and test B group.Rats of test groups were orally given live combined bifidobacterium and lactobacillus tablets for 7 days,respectively;rats of control group were given the same volume distilled water.On day 8,ticagrelor was orally administered to all rats.Blood samples were obtained at 0.08,0.25,0.50,1,1.5,2,3,4,6,8,12,24 h after ticagrelor administration.The plasma concentration of ticagrelor was determined by LC-MS/MS.The pharmacokinetic parameters were determined by DAS 2.1.1 software and the statistic analysis was processed with SPSS 21.0 software.Results The pharmacokinetic parameters of ticagrelor in the test A group,test B group and control group were as follows:AUC0-t were (6336.24 ± 1840.46),(4444.05± 1033.43) and (4469.32 ±928.47) ng· mL-1 · h-1;AUC0-∞ were (6841.98 ± 1975.95),(4656.66 ± 1083.78) and (4736.47 ± 897.42) ng · mL-1 · h;Cmax were (858.65 ± 275.98),648.81 ± 215.59) and (617.49 ± 168.95)ng · mL-1;t1/2 were (6.40 ± 2.18),(5.25 ± 1.39) and (5.68 ± 2.08) h;tmax were (0.88 ± 0.23),(0.90 ± 0.21) and (1.30 ± 0.59) h;clearance (CL) were (2.82±0.72) (4.07±0.99) and (3.95±0.91) L·h-1 ·kg-1;apparent volume of distribution (Ⅴ) were (26.07 ± 12.00),(31.45 ± 14.65) and (32.95 ± 14.17) L · kg-1.There were significant differences in AUC0-t,AUC0-∞,Cmax,tmax and CL between test A group and control group (P ＜ 0.05).On the contrary,no significant differences were observed on the pharmacokinetic parameters between test B group and control group.Conclusion Increased intestinal probiotics can increase the AUC0-t,AUC0-∞,and Cmax of ticagrelor,while decrease the CL of ticagrelor.

Objective To establish a HPLC method for the determination of paraquat in human plasma which can be used in clinical practice.Methods The plasma samples were processed with WCX solid phase extraction column and were determinated by RP-HPLC.The HPLC determination was conducted on a Diamonsil C18 column (250 mm × 4.6 mm,5 μm)with the mobile phase consisted of 0.1 mol · L-1 phosphate buffer (containing 80 mmol · L-1 sodium heptanesulfonate,pH 3.0)-acetonitrile (80:20),and the flow rate was 1.0 mL · min-1.The detection wavelength was set at 258 nm,and the column temperature was 25 ℃.Results The plasma concentration of paraquat had good linearity over the range of 20-5000 ng· mL-1 (r =0.999 9).The limit of detection for the paraquat was 20 ng· mL-1.The relative recovery rates of paraquat were 90.06％-104.50％.The absolute recovery rates were 97.66％-99.11％,respectively.The intra-day and inter-day precision (RSD) were all less than 4.00％.The paraquat plasma sample was all stable at the conditions of maintaining at 4 ℃ for 8 h,after freeze thaw cycle and keeping at-40 ℃ for 21 days.The method was applied to the clinical determination of paraquat and the calculation of severityindex of paraquatpoison (SIPP),which was helpful in the clinical treatment of patients with paraquat poison and to predict the prognosis.Conclusion A rapid,sensitive and accurate RP-HPLC method was established,which can be used in the determination of paraquat in human plasma and predict the effect of clinical treatment of patients with paraquat poison.

Objective To investigate the relationship between hyperuricemia and contrast-induced nephropathy (CIN) in patients with chronic kidney disease (CKD) undergoing percutaneous coronary intervention (PCI).Methods A total of 446 consecutive patients with CKD undergoing PCI in Guangdong general hospital were enrolled in this study.Patients were divided into hyperuricemic group (n =205) and normouricemic group (n =241).Hyperuricemia was defined as serum uric acid ＞ 420 μmol/L for male,＞357 μmol/L for female.CIN was defined as ≥44.2 μmol/L or ≥25％ increase from baseline Serum creatinine within 48-72 hours after contrast medium exposure,and that was not attributable to other causes.In hospital incidences of CIN and the major adverse cardiac events were compared between the two groups.The relationship between the incidence of CIN and hyperuricemia was evaluated by multivariate logistic regression analysis.Results CIN occurred in 16.6％ (74/446) of patients,and incidence of CIN was significantly higher in the hyperuricemic group than in the normouricemic group [23.9％ (49/446) vs.10.4％ (25/446),P =0.000].Patients who developed CIN had higher in hospital mortality [14.9％ (11/74) vs.1.3％ (5/372),P =0.000].Need for renal replacement therapy,acute heart failure,intraaortic balloon pump use and the hypotension after PCI were significantly higher in the hyperuricemic group compared with normouricemic group (P ＜ 0.01 or P ＜ 0.05).Multivariate analysis indicates that hyperuricemia (OR =1.9,95 ％ CI:1.1-3.5,P =0.037),age ＞ 75 years (OR =3.2,95 ％ CI:1.8-5.7,P =0.000),emergent PCI (OR =2.9,95 ％ CI:1.6-5.1,P =0.000) and anemia (OR =2.1,95 ％ CI:1.2-3.8,P =0.012) were predictors of CIN in patients with CKD.Conclusion Hyperuricemia is the independent risk predictor of CIN in patients with CKD undergoing PCI.