Objective To investigate the influence of di-(2-ethylexyl) phthalate (DEHP) on cell apoptosis and expression of Bcl-2 and bax in cultured human first trimester cytotrophoblasts. Methods Human first trimester cytotrophoblasts were cultured with DEHP at concentration of 0, 25, 50, 100 μmol/Lfor 24 hours. Cell apoptosis was detected by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) and flow cytometer method. The expression of apoptosis-associated genes, including Bcl-2 and bax, were detected by reverse transcription (RT)-PCR in cultured cytotrophoblast cells. The protein expression of Bcl-2 and bax in cytotrophoblast cells was measured by western blot. Results (1) The expression of Bcl-2: when incubated with DEHP at concentration of 0, 25,50 and 100 μmol/L, the expression of Bcl-2 were 1.00 ± 0.05, 1.03 ± 0.04, 1.04 ± 0.03, 1.04± 0.04,which did not show statistical difference ( P > 0.05 ). The expression of Bcl-2 protein were 0.11 ± 0.02,0.11 ±0.04, 0.12±0.02, 0.12 ±0.03, which also didn't reach statistical difference (P>0.05). (2)The expression of bax: when incubated with DEHP at concentration of 50 and 100 μmol/L, the expression of bax protein were 0.63 ± 0.04 and 0.81 ± 0.04, which were significantly higher than 0.23 ± 0.05 with DEHP at 0 μmol/L (P < 0.05). The expression of bax mRNA were 0.96 ± 0.04 and 1.02 ± 0.04, which was significantly higher than 0.81 ±0.05 with DEHP at 0 μmol/L (P < 0.05). (3) Apoptosis: when incubated with DEHP at concentration of 50 and 100 μmol/L for 24 hours, the apoptotic cell ratio were ( 18.8 ± 2.6) % and ( 20.3 ± 2.0) % by annexin V-FITC/PI staining, which were significantly higher than (10.6±1.4)% at 0 μmol/L and (18.1 ±4.6)% and (19.5 ±1.2)% by TUNEL staining, which were significantly higher than ( 11.2 ± 3.1 ) % at 0 μmol/L of DEHP (P < 0.05). Conclusion DEHP could induce apoptosis of cytotrophoblast cells by increasing bax gene expression, but had no effect on Bcl-2 expression.

This study is to analyze and identify the water soluble components of water buffalo horn (Bubali Cornu, WBH), and also establish a method for investigating these components. Shotgun proteomic analysis identified proteins in WBH aqueous extraction: keratin, collagen, desmoglein, etc. Ultrafiltration and LC-MS/MS were used to separate and identify the peptides in WBH aqueous extract, as a result, identified peptides were mainly derived from nonspecific degradation products of keratin and collagen, which including C-terminal peptides and non C-terminal peptides. Hypoxanthine, uridine, guanosine, and adenosine were identified by comparing with the standards. The strategy in present study could be used in analyzing water soluble components of animal horn derived TCM. It provides a reference for investigation of the material basis of animal horn derived TCM.
Animals ; Buffaloes ; Chromatography, Liquid ; Guanosine ; Horns ; chemistry ; Hypoxanthine ; Mass Spectrometry ; Medicine, Chinese Traditional ; Peptides ; Proteomics ; Tandem Mass Spectrometry ; Uridine

Objective To evaluate morphological structure of uterosacral ligament (USL) and cardinal ligament (CL) in patients with severe pelvic organ prolapse (POP) by MRI technology, and to analysis and discuss its clinical significance. Methods From November 2013 to February 2014 in Peking University People′s Hospital, 26 elderly patients withⅢ-Ⅳdegree of POP were selected as the POP group and 18 healthy elderly volunteers were selected as the control group during the same period. Pelvic MRI examination were performed in the two groups. The morphological characteristics of left and right side of the uterosacral-cardinal ligament on MRI and the attachment site of the starting and ending points between two group were described and compared. Results In POP group, 25 cases of left USL starting point were located in the sacrospinous ligament/coccygeal muscle complex [58% (15/26)] or coccygeal muscle [38%(10/26)], ending point were located in the cervix and vagina [58%(15/26)] or cervix [38%(10/26)];24 cases of right USL starting point were located in the sacrospinous ligament/coccygeal muscle complex [31%(8/26)]or coccygeal muscle [62%(16/26)], 26 cases of right USL ending point were located in the cervix and vagina [62% (16/26)] or cervix [38% (10/26)]; the left and right CL in the POP group and the control group were both from the sacroiliac joint at the top of the greater sciatic foramen from the ipsilateral pelvic side wall;1 case (4%, 1/26) of left CL in the POP group completely connected to the bladder, 10 cases (38%, 10/26) partly connected to the bladder;14 cases (54%, 14/26) of right CL partly connected to the bladder, the rest ending points of left and right CL were located in cervix and (or) vagina. In the control group, 17 cases of left USL starting point were located in the sacrospinous ligament/coccygeal muscle complex (10/18) or coccygeal muscle (7/18), ending point were located in the cervix and vagina (12/18) or cervix (6/18);18 cases of right USL starting point were located in the sacrospinous ligament/coccygeal muscle complex (10/18) or coccygeal muscle (8/18), ending point were located in the cervix and vagina (13/18) or cervix (5/18);8 cases (8/18) of left CL partly connected to the bladder;15 cases (15/18) of right CL partly connected to the bladder, the rest ending points of left and right CL were located in cervix and (or) vagina. There was no significant difference between the two groups on the starting and ending points (P>0.05). Conclusions The observation of MRI could be consistent with the clinical anatomy on the starting and ending points, direction of travel in the uterosacral-cardinal ligament. The starting and ending points of the left and right side USL and the ending points of the left and right side CL are not completely symmetrical, the variation degree is large, some CL could be completely or partly inserted to the bladder.

The aim of the paper is to reveals the variations of Dendrobium officinale amino acids in different strains and parts for breeding excellent varieties, and providing scientific basis for the expanding of medicinal or edible parts. The contents of 17 amino acids in 11 strains of D. officinale were determined by hydrochloric acid hydrolysis method. The total amino acids content of leaves was from 6.76 to 7.97 g per 100 g, and the stems was from 1.61 to 2.44 g per 100 g. As the content of amino acids in leaves was significantly higher than that of stems, and the composition was close to the ideal protein standard proposed by FAO/WHO. The leaves of D. officinale had the good prospect for the development of functional foods. The 9 x 66 strain which with high yield and polysaccharide content had the highest amino acids content both in stems and leaves, indicated crossbreeding could improve the quality of varieties. Compared the amino acids content of D. officinale in two main harvest periods, the harvest time has a significant impact on amino acids content of D. officinale. This study demonstrates that the harvesting time of D. officinale stems is suitable for leaves as well, which is the period before bolssom.
Amino Acids ; chemistry ; Dendrobium ; chemistry ; classification ; Drugs, Chinese Herbal ; chemistry ; Phylogeny ; Plant Leaves ; chemistry ; classification

The stability of melittin in different solvents (water, deoxygenated water, physiological saline, PBS, 50% ethanol, ethanol, glycerol)was studied and the results showed that the stability of melittin is not influenced by light, temperature and pH in 50% ethanol, which melittin can be completed dissolved when compared with ethanol and glycerol, in such, 50% ethanol was chosen as solvent storage when measured content of melittin. Then the effect of different concentrations of PBS, the pH of PBS and rat skin ho- mogenates were tested, and the results showed that melittin was degraded rapidly at low concentration solution and low ionic strength. Increasing pH of PBS and rat skin homogenate can accelerate the degradation of melittin. These researches provide an experimental ba- sis for further study of melittin.
Animals ; Drug Stability ; Ethanol ; chemistry ; Hydrogen-Ion Concentration ; Melitten ; chemistry ; Rats ; Skin ; drug effects ; Solvents ; chemistry ; Temperature

Objective Through the detection of the levels of serum betatrophin,anthropometric and biochemical indices,to determine the alteration of betatrophin levels in patients with polycystic ovary syndrome (PCOS) and the relationship between betatrophin levels and metabolic indexes such as 25-hydroxy vitamin D3 [25 (OH) D3].Methods The study group was composed of 24 women with PCOS and 20 age-natched healthy women as controls.Following general physical examination of the subjects,anthropometrie measurements were performed (height,weight,waist circumference,hip circumference).Body mass index (BMI) and waist-hip ratio were calculated;25 (OH) D3,Ca2+,fasting blood glucose,fasting insulin,triglycerides,high-density lipoprotein cholesterol (HDL-C),low-density lipoprotein cholesterol (LDL-C) were also measured and homeostasis model assessment of insulin resistance (HOMA-IR) were evaluated in all subjects.Serum betatrophin levels were examined with an enzyme-linked immunosorbent assay (ELISA).Using independent sample T test to compare the differences between groups,the linear correlation analysis was performed to study the correlation of betatrophin with anthropometric and biochemical indices,the influencing factors of betatrophin were analyzed by multiple linear regression.Results Circulating betatrophin levels [(0.341±0.034) ng/ml vs.(0.810±0.162) ng/ml,t=4.271,P＜0.001] and HDL-C [(1.014±0.321) mmol/L vs.(1.419±0.287) mmol/L,t=2.218,P＜0.05] significantly decreased in women with PCOS compared with controls,and BMI [(27.691±4.392) kg/m2 vs.(23.310±4.781) kg/m2,t=-2.073,P＜0.05],fasting blood glucose [(5.950±0.411) mmol/L vs.(4.883±0.314) mmol/L,t=-2.142,P＜ 0.05],HOMA-IR [(4.946 ± 0.741) vs.(3.387± 0.397),t=-2.493,P＜0.05],triglycerides [(1.510± 0.848) mmol/L vs.(1.037±0.402) mmol/L,t=-2.223,P＜0.05],LDL-C [(3.431±0.479) mmol/L vs.(2.396±0.435) mmol/L,t=-2.433,P＜0.05] were obviously increased.Moreover,there was a postive correlation between betatrophin and HOMA-IR (r=0.425,P＜0.05) as well as 25(OH)D3 (r=0.577,P＜0.05) and Ca2+ (r=0.448,P＜0.05),while the betatrophin was negatively related to BMI (r=-0.451,P＜0.05),triglycerides (r=-0.454,P＜0.05),LDL-C (r=-0.551,P＜0.05).Circulating betatrophin levels were higher when 25 (OH) D3 levels were beyond 35 nmol/L in PCOS patients [(0.539±0.092) ng/ml vs.(0.199±0.031) ng/ml,t=3.072,P＜0.001],and the multiple linear regression analysis showed that the main factors affecting the levels of betatrophin were BMI (OR=-0.260),HOMA-IR (OR=0.218),25(OH)D3 (OR=0.238),and glycerol (OR=-0.162).Conclusion It is speculated that betatrophin may be a valuable predictive factor of PCOS because betatrophin is closely associated with insulin resistance,lipid metabolism disorders and the lack of 25-hydroxy vitamin D3 in PCOS patients.

This paper revealed the accumulation regularity of polysaccharides and alcohol-soluble extracts contents of Dendrobium officinale leaves, which have provided basis for the development and utilization of the leaves. The polysaccharides and alcohol-soluble extracts contents of three D. officinale strains leaves collected in different growing periods were determined by phenol-sulfric acid method and hot-dip method respectively. The results showed that the content of polysaccharides in leaves was 4.45% -12.17%, and was about a quarter in stems. The alcohol-soluble extracts content in leaves was 7.45% - 29.34%, and was 1.5 times that of stems. The quality variation of polysaccharides in leaves was closely related to the phenophase. The leaves with lower level of metabolism in three stages: winter, early germination stage and deciduous period, which led to lower content of polysaccharides. The leaves at the vigorous growth stage with higher content of polysaccharides. The alcohol-soluble extracts were closely associated to the formation and germination of buds. The content of alcohol-soluble extracts peaked before sprout, and promoted the growth of new shoots.
Dendrobium ; chemistry ; metabolism ; Drugs, Chinese Herbal ; analysis ; metabolism ; Plant Leaves ; chemistry ; metabolism ; Polysaccharides ; analysis ; metabolism ; Seasons

AIMTo apply stochastic resonance algorithm (SRA) to quantitative analysis of weak chromatographic signal, which was embedded in the noise.

METHODSBased on the theory of stochastic resonance (SR), a simple and effective SRA has been established to improve analytical detection limits of chromatographic analysis, which apply to enhance the signal to noise ratio by the optimization of the parameters and Runge-Kutta method, was established. The method was used to quantitative analysis of phenazopyridine in human plasma by HPLC/UV. Meanwhile this method is compared with HPLC/MS.

RESULTSBy experimental chromatographic data sets, an excellent quantitative relationship between concentrations of phenazopyridine and their responses had been obtained. The concentration of phenazopyridine in plasma determined by HPLC/UV with SRA and HPLC/MS showed that there was no significant difference (P > 0.05) between the two methods.

CONCLUSIONThe new method was feasible.

Chromatography, High Pressure Liquid ; methods ; Humans ; Mass Spectrometry ; Noise ; Phenazopyridine ; blood ; Sensory Thresholds ; Spectrophotometry, Ultraviolet ; Stochastic Processes

Objective To establish the finite element model of uterosacral ligament (USL) and cardinal ligament (CL) and analyze the stress distribution and deformation with USL and CL under different working conditions. Methods Patients with stage Ⅲ-Ⅳpelvic organ prolapse (POP) and healthy female volunteers were selected for research subject, and divided into anterior uterus group and posterior uterus group. Two POP patients and two volunteers were selectd into the anterior uterus group and posterior uterine group respectively. Pelvic MRI scan was performed in two groups. Based on the original MRI data sets, the finite element model of USL and CL was constructed by using the software such as the Mimics, and the stress distribution and deformation of USL and CL were simulated. Results Under the premise of the elastic modulus fixed and three different working conditions such as 60 cmH2O, 99 cmH2O and 168 cmH2O (1 cmH2O=0.098 kPa) with abdominal pressure generated by maximum Valsalva maneuver, according to the present conditions and the simulation, the trend was analyzed: the stress and deformation of the uterus, anterior vaginal wall, USL and CL in two groups were mainly distributed in the middle and lower part of the anterior vaginal wall or the ligament and the cervix-vagina junction, the maximum stress and the maximum displacement were mainly concentrated in the lower region of the anterior vaginal wall. With increasing of abdominal pressure generated by the maximum Valsalva maneuver, the maximum stress values of the POP patient in anterior uterus group under three different working conditions were: 0.027 9, 0.046 0, 0.078 0 MPa, and the maximum displacement values were: 9.145 5, 15.090 0, 25.607 0 mm. The maximum stress values of the volunteer in anterior uterus group under three different working conditions were:0.012 6, 0.020 8, 0.035 3 MPa, and the maximum displacement values were: 1.816 7, 2.997 5, 5.086 7 mm. The maximum stress values of the POP patient in posterior uterine group under three different conditions were: 0.069 4, 0.114 6, 0.194 5 MPa, and the maximum displacement values were:11.658 0, 19.236 0, 32.643 0 mm. The maximum stress values of the volunteer in posterior uterus group under three different working conditions were:0.009 1, 0.015 1, 0.025 6 MPa, and the maximum displacement values were:2.581 6, 4.259 6, 7.228 4 mm. The maximum stress values and the maximum displacement values were all increased with increasing of abdominal pressure in the two groups. The maximum stress values and the maximum displacement values of the POP patients were greater than those of volunteers. Under different working conditions, the maximum stress values and maximum displacement values of the posterior uterus POP patient were all greater than those of the anterior uterus POP patient. Conclusions The finite element model of USL and CL is completely based on the MRI technology and the model is real and reliable. The increase of abdominal pressure will produce a larger stress and deformation of USL and CL, which is one of the reasons causing the injury of the ligament.

OBJECTIVETo evaluate the therapeutic effect of Compound Xuanju Capsule (CXC) on autoimmune prostatitis in rat models.

METHODSSixty healthy male Wistar rats were randomly divided into five groups of equal number: blank control, low-concentration purified prostate protein (low-conc PPP), low-conc PPP + CXC treatment, high-concentration PPP (hi-con PPP), and hi-conc PPP + CXC treatment. Autoimmune prostatitis models were established by intragastric administration of PPP solution at 15 mg/ml (low concentration) and 80 mg/ml, respectively. At 30 days after modeling, the rats in the blank control and low-conc and hi-conc PPP model groups were treated with normal saline, and those in the other two groups with CXC at a daily dose of 0.068 g/ml. At 30, 45, and 60 days, all the animals were sacrificed for observation of pathological changes in the prostate tissue and determination of the levels of IL-8, IL-10, and TNF-alpha in the serum.

RESULTSCompared with the PPP models, the hi-conc PPP + CXC group showed significantly reduced levels of IL-8 and TNF-alpha in the serum at 45 days ([148.54 +/- 17.23] and [62.14 +/- 5.59] pg/ml vs [100.77 +/- 11.08] and [32.63 +/- 2.91] pg/ml, P < 0.05) and at 60 days ([143.69 +/- 17.28] and [59.38 +/- 5.50] pg/mlvs [95.77 +/-10.53] and [29.63 +/- 2.66] pg/ml, P < 0.05), and so did the low-cone PPP + CXC group at 45 days ([128.47 +/- 12.21] and [40.43 +/- 3.64] pg/ml vs [111.76 +/- 10.07] and [35.44 +/- 3.17] pg/ml, P < 0.05) and at 60 days ([131.07 +/- 10.93] and [43.34 +/- 3.91] pg/ml vs [97.46 +/- 8.75] and [30.44 +/- 2.75] pg/ml, P < 0.05). The serum level of IL-10 was remarkably elevated in the hi-cone PPP + CXC group as compared with that of the PPP models at 45 and 60 days ([189.14 +/- 16.78] and [184.14 +/- 15.89] pg/ml vs [230.48 +/- 29.96] and [248.48 +/- 31.03] pg/ml, P < 0.05), and so was it in low-cone PPP + CXC group ([223.14 +/- 17.87] and [224.14 +/- 17.93] pg/ml vs [231.42 +/- 23.18] and [249.42 +/- 24.97] pg/ml, P < 0.05). Pathological examination revealed morphological damages to the prostate tissue and infiltration of inflammatory cells in the model rats, but no obvious changes in the normal controls. At 15 days of treatment, the rats in the PPP + CXC group showed enlarged prostate glandular cavity, mild proliferation of epithelial cells, no obvious infiltration of inflammatory cells in the interstitial tissue, and a few visible fibrous tissues under the light microscope.

CONCLUSIONCompound Xuanju Capsule is efficacious on autoimmune prostatis in rats by reducing inflammatory changes in the prostate tissue and improving the expression of inflammatory factors.

Animals ; Autoimmune Diseases ; blood ; chemically induced ; drug therapy ; Capsules ; Interleukin-10 ; blood ; Interleukin-8 ; blood ; Male ; Prostatic Hyperplasia ; pathology ; Prostatic Secretory Proteins ; Prostatitis ; blood ; chemically induced ; drug therapy ; Random Allocation ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; blood