Effect of short-term nutritional supplementation on hormone concentrations in ovarian follicular fluid and steroid regulating gene mRNA abundances in granulosa cells of ewes.

This study was conducted to investigate effects of short-term nutritional supplementation on concentrations of reproductive hormones in follicular fluid and on mRNA abundance in granulosa cells (GCs) during the luteal phase of ewes. Eighteen ewes were randomly assigned to treatment or control groups (n = 9, each group). All the ewes were subjected to estrous synchronization using a Controlled Intravaginal Releasing Device (CIDR). From the second to the eleventh day of estrous synchronization, ewes were fed a diet with a relatively greater (treatment group) or a maintenance (control group) energy content. Samples of follicular fluid and GCs were collected from antral follicles of estrous ewes after CIDR removal. The results indicate mean FSH concentration of fluid in small follicles and mean LH concentrations of fluid in large follicles of the ewes in the treatment group were greater (P < 0.05) than that of ewes in the control group. Follicular fluid E2 concentrations were directly related (P < 0.05) to follicular diameter. The ewes of the treatment group had greater (P < 0.05), compared with the control group, abundances of Follicle Stimulating Hormone Receptor (FSHR) in small and medium follicles, and (P<0.05) Luteinizing Hormone Receptor (LHR), Steroid Acute Regulatory Protein (STAR), cytochrome P450 (CYP17A1, CYP19A1) enzyme and Estrogen Receptor (ESR1) in large follicles. The results of this study provide evidence for a potential mechanism by which short-term nutritional supplementation improves follicular development possibly because of increased expression of steroid synthesis-regulating genes in large follicles.

[Low power seven-step two-lobe holmium laser enucleation of the prostate technique for surgical treatment of benign prostatic hyperplasia].

OBJECTIVE: To evaluate the safety and efficacy of the seven-step two-lobe holmium laser enucleation of the prostate (HoLEP) technique with low power laser device, and to introduce the detailed operating procedures, key points, short-term outcomes of this modified HoLEP technique. METHODS: From March 2016 to November 2017, 90 patients underwent HoLEP in Peking University Third Hospital. The patients were divided into two groups: high-power group (32 patients) were performed with traditional Gilling's three-lobe enucleation using high power (90 W) laser; Low-power group (58 patients) were performed with seven-step two-lobe enucleation using low power (40 W) laser. The main steps of the low power seven-step two-lobe HoLEP phase included: (1) The identification of the correct plane between adenoma and capsule at 5 and 7 o'clock laterally to the veru montanum; (2) The connection of the bilateral plane by making a adenoma incision at the proximal point of veru montanum; (3) The extension of the dorsal plane under the whole three lobes between adenoma and capsule towards the bladder neck; (4) The separation of the middle lobe from two lateral lobes by making two retrograde incisions separately from apex 5 and 7 o'clock towards the bladder neck; (5) The enucleation of the middle lobe adenoma by extending the dorsal plane through into the bladder; (6) The prevention of the apex mucosa by making a circle incision at the apex of the prostate; (7) The en-bloc enucleation of the two lateral lobe adenomas by extending the lateral and ventral plane between adenoma and capsule from 5 and 7 o'clock to 12 o'clock conjunction and through into the bladder. RESULTS: The mean patient age was (66.25±5.37) years vs. (68.00±5.18) years; The mean body mass indexes were (24.13±4.06) kg/m2 vs. (24.57±3.50) kg/m2; The mean prostate specific antigen values were (3.23±2.47) µg/L vs. (6.00±6.09) µg/L; The average prostatic volumes evaluated by ultrasound was (49.03±20.63) mL vs. (67.55±36.97) mL. There was no significant difference between the two groups. Furthermore, there were no significant differences in terms of perioperative and follow up data, including operative time; enucleation efficiencies; hemoglobin decrease; blood sodium and potassiumthe change postoperatively; catheterization duration and hospital stay; the international prostate symptom scores and quality of life scores pre- and post-operatively. There was 1 transurethral resection of the prostate (TURP) conversion in high-power group and 1 transfusion in low-power group during the operations. The follow-up one month after operation showed no severe stress incontinence in both the groups, whereas 3 cases ejaculatory dysfunctions in high-power group versus 1 case in low-power group were observed; Other surgeryrelated complications included: 2 cases postoperative hemorrhage (Clavien II and Clavien IIIb) in high-power group, 2 cases postoperative temperature more than 38 °C (Clavien I) and 1 case dysuria following catheter removal (Clavien I) in low-power group. CONCLUSION: Low power laser device can be applied safe and effectively for HoLEP procedure using the seven-step two-lobe HoLEP technique. The outcomes comparable with high power laser HoLEP can be achieved.

[Clinical study of "point-line-surface-volume" four dimensional holmium laser enucleation of prostate for benign prostatic hyperplasia].

Objective: To investigate application value of "point-line-surface-volume" four dimensional holmium laser enucleation of prostate (HoLEP) in the treatment of benign prostatic hyperplasia (BPH). Methods: From December 2016 to November 2017, the clinical data of 60 cases of BPH with the treatment of "Point-line-surface-volume" four dimensional HoLEP at Department of Urology, Peking University Third Hospital were analyzed retrospectively. The age was (73.8±8.0) years (range: 60 to 96 years), body mass index was (23.2±3.6) kg/m(2) (range: 14.9 to 31.1 kg/m(2)), volume of prostate was (64.5±36.9) ml (range: 15.5 to 197.9 ml). All the cases were operated by the same chief surgeon expert in endoscopic surgery. First of all, taking verumontanum as the anchor point, crossing point between the outer 45° line of verumontanum and middle line of the lateral lobe and verumontanum was made as the breakthrough point. The "blasting and paddling" method was used to find prostatic surgery capsule, and the two sides of the capsule plane were connected in front of the verumontanum. Secondly, the plane was maintained by the "fan tunnel" method, and the middle lobe and the lateral lobes were divided at the 5 o'clock and 7 o'clock positions. The glands of the middle lobe were first removed and pushed into the bladder. Then, on the anteroinferior inclined coronal plane through 12 o'clock point right above of verumontanum, the urethral mucosa was cut apart in a semi-curved shape in advance. Bilateral semi-curved incision confluenced above the verumontanum, and extend to the bladder neck to form an inverted Y-shaped groove which completely divided the bilateral lobes. Finally, the bilateral lobes were enucleated respectively, and glands in the bladder were removed by tissue morcellator. Clinical data included operative time, hemoglobin decrease, catheterization duration, postoperative hospital stay, preoperative and postoperative international prostate symptom score and quality of life (QOL) score. The efficiency of enucleation was calculated as prostatic volume divided by enucleation time (not including morcellation time). The efficiency of morcellation was calculated as prostatic volume divided by morcellation time. Paired t-test was used to compare the indexes before and after surgery. Results: All the operations of 60 cases were sumlessful, one of which prostate tissue was removed by conversion to transurethral resection prostate due to malfunction of tissue morcellator. The operative time was (115.2±52.9) minutes (range: 25 to 276 minutes). The enucleation efficiency was (0.81±0.35) ml/minutes (range: 0.17 to 1.58 ml/minutes). The morcellation efficiency was (6.60±4.28) ml/minutes (range: 0.89 to 17.42 ml/minutes). The hemoglobin was decreased by (15.9±12.3) g/L (range:-10 to 57 g/L). Meanwhile, catheterization duration was (5.2±2.9) days (range: 0.8 to 19.8 days), and postoperative hospital stays were (5.0±1.5) days (range: 1.9 to 11.9 days). Preoperative and postoperative IPSS scores (10.92±6.98 vs. 23.37±7.49, t=10.357, P=0.000) and QOL scores (1.75±1.62 vs. 4.53±1.47, t=9.373, P=0.000) were significantly different. Postoperative complications included: 4 cases of fever (greater than 38.5 ℃), 1 case of acute epididymitis, 1 case of bladder mucosal injury, and 1 case of active bleeding need blood transfusion therapy. Conclusion: "Point-line-surface-volume" four dimensional HoLEP is an effective and safe minimally invasive method in the treatment of BPH.

[Transurethral flexible ureteroscopic holmium laser resection for tumors of renal pelvis: 6 cases report and literature review].

OBJECTIVE: To summarize the experience of flexible ureteroscopic holmium laser resection in treatment of renal pelvic carcinoma and to evaluate its value in treatment of renal pelvic carcinoma. METHODS: The clinical data of 6 patients with renal pelvic carcinoma treated in Peking University Third Hospital from January 2015 to January 2017 were retrospectively analyzed. The 6 patients were treated by the same experienced urologist and by flexible ureteroscopic holmium laser resection of renal pelvic tumors under general anesthesia. Regarding the intensity of the holmium laser, 10-30 W was generally used with settings of 0.5-1.5 J and 10-20 Hz. In general, a 200 µm end-firing holmium laser fiber was used. Narrow-band imaging (NBI) technique was applicated to search for tumors and check whether the excision was satisfactory. Routine "second flexible ureteroscopy" was performed after 4-6 weeks, and suspected lesions were referred for a biopy, then vaporized and cauterized. The ureteroscopy was examined every 6 months after operation, and color Doppler ultrasound, computed tomography urography (CTU) or magnetic resonance urography (MRU) were performed at the same time. The urine tumor cells were examined for 3 days before the operation, and the urine tumor markers, such as urinary nuclear matrix protein 22 (NMP22) were tested. For cases with highisk urothelial carcinoma and normal renal function, and 6 cycles of systemic adjuvant chemotherapy were performed after operation. RESULTS: All of the cases were successfully treated. The data were as follows: the operation time 77.5 min (45-115 min), the blood loss 10 mL (5-20 mL), and hospital stay after surgery 3 days (2-5 days). After 13-34 months' followp, two patients had recurrent tumor recurrence and underwent resection operation. Two patients received systemic adjuvant chemotherapy after operation. Case 5 was histopathologically high grade urothelial carcinoma, and 6 cycles of systemic chemotherapy were given after operation. Local recurrence occurred during chemotherapy, and then endoscopic operation was performed, and no recurrence occurred in the follow-up for 12 months after reoperation. In case 6, the pathology was low grade urothelial carcinoma, but the case was multiple tumors in the right renal calyx and the lower calyx. Then 6 cycles of systemic chemotherapy were given, and no recurrence was found in the followp for 13 months. CONCLUSION: Transurethral flexible ureteroscopic holmium laser resection is relatively safe for the treatment of renal pelvic carcinoma. It is suitable for special cases of solitary kidney and renal dysfunction, as well as for patients with low risk urinary tract epithelial tumors, but the recurrence rate is high, and the indications need to be strictly controlled. Patients with high-risk urothelial carcinoma who underwent endoscopic resection are advised to receive systemic adjuvant gemcitabine and cisplatin (GC) regimen after surgery, in order to increase the overall survival rate. Systemic chemotherapy combined with endoscopic operation may become a new treatment for upper tract urothelial carcinoma (UTUC).

Effect of dietary arginine and N-carbamoylglutamate supplementation on reproduction and gene expression of eNOS, VEGFA and PlGF1 in placenta in late pregnancy of sows.

The objectives of this study were to investigate the potential mechanisms of dietary arginine (Arg) and N-carbamoylglutamate (NCG) supplementation on reproductive performance of sows. Twenty-seven crossbred (Landrace×Large White) sows with similar body weight and parity at day (90±1) of gestation were assigned randomly into 3 groups (n=9) control group, Arg group, NCG group, and fed with the following diets: a control diet, and the control diet supplemented with 1.0% Arg or 0.1% NCG. Litter size was recorded. Blood samples were obtained for biochemical analyses. Placenta chorioallantoic membrane tissue collected immediately after birth to preserve in RNA stabilizer for mRNA analysis of endothelial nitric oxide synthase (eNOS), endothelial growth factor a (VEGFA) and placenta growth factor 1 (PlGF1) by real time-PCR. The results showed that compared with the control group, the average birth weight of all piglets born alive were 16.2% and 14.3% higher in the Arg and NCG groups (P<0.05), respectively; plasma VEGFA was higher in the Arg group (P<0.05). The expression of VEGFA in the allantochorion tissue of the NCG-supplemented group was higher (P<0.01), and tended to be higher in the Arg-supplemented group (0.05

Effects of dietary L-arginine or N-carbamylglutamate supplementation during late gestation of sows on the miR-15b/16, miR-221/222, VEGFA and eNOS expression in umbilical vein.

Placental vascular formation and blood flow are crucial for fetal survival, growth and development, and arginine regulates vascular development and function. This study determined the effects of dietary arginine or N-carbamylglutamate (NCG) supplementation during late gestation of sows on the microRNAs, vascular endothelial growth factor A (VEGFA) and endothelial nitric oxide synthase (eNOS) expression in umbilical vein. Twenty-seven landrace×large white sows at day (d) 90 of gestation were assigned randomly to three groups and fed the following diets: a control diet and the control diet supplemented with 1.0% L-arginine or 0.10% NCG. Umbilical vein of fetuses with body weight around 2.0 kg (oversized), 1.5 kg (normal) and 0.6 kg (intrauterine growth restriction, IUGR) were obtained immediately after farrowing for miR-15b, miR-16, miR-221, miR-222, VEGFA and eNOS real-time PCR analysis. Compared with the control diets, dietary Arg or NCG supplementation enhanced the reproductive performance of sows, significantly increased (P<0.05) plasma arginine and decreased plasma VEGF and eNOS (P<0.05). The miR-15b expression in the umbilical vein was higher (P<0.05) in the NCG-supplemented group than in the control group. There was a trend in that the miR-222 expression in the umbilical vein of the oversized fetuses was higher (0.05

Induction of cytochrome P450s by terpene trilactones and flavonoids of the Ginkgo biloba extract EGb 761 in rats.

1. Ginkgo biloba is one of the most popular herbal medicines worldwide due to its memory-enhancing and cognition-improving effects. The current study was designed to investigate the effects of five major constituents (bilobalide, ginkgolide A, B, quercetin, and kaempferol) in the standardized G. biloba extract EGb 761 on various cytochrome P450s (CYPs) in rats. 2. The activity of CYP450 was measured by the quantification of six metabolites from multiple cytochrome P450 probe substrates using a validated liquid chromatography coupled with tandem mass spectrometry detection (LC-MS/MS) method. The levels of messenger RNA (mRNA) and protein of various CYPs were determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting analysis, respectively. 3. Bilobalide significantly induced the activity, protein, and mRNA expression of CYP3A1 and 1A2, and increased CYP2E1 activity and CYP2B1/2 protein expression in a dose-dependent manner. 4. Ginkgolide A, B, quercetin, and kaempferol did not affect CYP3A1, but induced CYP1A2 in a dose-dependent manner. EGb 761 and the five individual constituents had no effects on rat CYP2D2, 2C11 and 2C7. 5. The results indicate that bilobalide, and to a lesser extent ginkgolide A, B, quercetin, and kaempferol, play a key role in the effects of EGb 761 on CYP induction. Further study is needed to elucidate the mechanism of CYP3A induction by EGb 761 and bilobalide.

Isolation and identification of a novel flavonoid from Penthorum chinense P.

A novel flavonoid, pinocembrin-7-O-[3''-O-galloyl-4'',6''-hexahydroxydiphenoyl]-beta-glucose (1) was isolated from the whole plant of Penthorum chinense P., along with four known compounds, pinocembrin-7-O-beta-glucoside, quercitrin, quercetin-3-O-rhamnoside and gallic acid. The structures were established by spectroscopic analysis.

[Clinical study on effect of recombinant roasted licorice decoction combined with low-dose glucocorticoids in treating idiopathic thrombocytopenic purpura].

OBJECTIVE: To explore the clinical effect of recombinant roasted licorice decoction (RRLD) combined with low-dose glucocorticoids (GC) in treating idiopathic thrombocytopenic purpura (ITP). METHODS: Forty-three patients were divided semi-randomly into the treated group (n = 21) and the control group (n = 22), the former was treated with RRLD plus low-dose GC, and the latter was treated mainly with normal dosage of GC. RESULTS: The total effective rate of the treated group was better than that of the control group, the difference between these two groups was significant (P < 0.05). After treatment the platelet count in the treated group arose from (36.29 +/- 9.70) x 10(9)/L to (95.14 +/- 18.26) x 10(9)/L, while in the control group from (37.31 +/- 9.29) x 10(9)/L to (77.73 +/- 21.96) x 10(9)/L, the RRLD was superior to nat of GC in raising platelet count, the difference between them was also significant (P < 0.01). The treated group was lower obviously than the control group in adverse reaction (P < 0.01). CONCLUSION: The efficacy of RRLD plus low-dose GC is reliable in treating ITP, with less adverse reaction.

[A comparison of chemical composition and bioactivity of polypeptides from velvet antlers of Cervus nippon Temminck and Cervus elaphus Linnaeus].

OBJECTIVE: To compare the chemical composition and bioactivity of polypeptides(PPs) isolated from velvet antlers of sika deer (Cervus nippon Temminck) and red deer (Cervus elaphus Linnaeus). METHOD: The two kind of polypeptides were isolated from the above mentioned velvet antlers with same technology. The chemical composition was determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and MALDI-TOF mass spectrometry. Stimulant activity of cells proliferation was measured by [3H] TdR incorporation into DNA. RESULT: The graphs of SDS-PAGE and MALDI-TOF MS of velvet antler polypeptides (VAPPs) from Chinese and New zealand red deer were very similar, but there were obvious difference in respect of graph between sika deer and red deer. VAPPs 25-50 mg.L-1 showed marked proliferation-promoting activity for rabbit costed chondrocytes, either sika deer or red deer. However, the activity of sika deer VAPPs 12.5 mg.L-1 for epidermal cells was weaker than that of red deer (12.5 mg.L-1). CONCLUSION: The chemical property and bioactivity of VAPPs from sika deer and red deer are significantly different.

[Direct optical resolution of acidic biphenyl drugs by high performance liquid chromatography on tris(3,5-dimethylphenylcarbamate) of cellulose].

A chiral stationary phase was prepared by coating cellulose-tris(3, 5-dimethylphenylcarbamate) onto aminopropylated silica gel. A series of enantiomeric acidic biphenyl drugs were directly resolved on the chiral stationary phase (CSP) by normal-phase high performance liquid chromatography (HPLC). A hexane-2-propanol eluting system containing 1% of trifluoroacetic acid was used as mobile phase. Efficient optical resolution of the acidic biphenyl drugs has been attained. The factors that influence chiral discrimination such as structural characeristic of the samples and mobile phase were investigated. An interaction model between the stationary phase and the samples was discussed. The results showed that efficient optical resolution of racemic carboxylic acids could be attained by normal-phase HPLC on CSP using a hexane-2-propanol eluting system containing 1% of trifluoroacetic acid.

Velvet antler polypeptides promoted proliferation of chondrocytes and osteoblast precursors and fracture healing.

AIM: To study the effects of velvet antler (VA) total polypeptides (VATP) and VA polypeptides, VAP-A, VAP-B, and VAP-C on proliferation of chondrocytes and osteoblast precusors. METHODS: Chondrocytes (rabbit and human fetus) and osteoblast precusors (chick embryo) were incubated in the culture medium containing VATP or VAP-A, VAP-B, and VAP-C. [3H]TdR incorporation into DNA was measured. Fracture healing-promoting action of VATP was determined in rats. RESULTS: VATP 50-200 mg.L-1 and VAP-B 12.5, 25, and 50 mg.L-1 showed most marked proliferation-promoting activity for rabbit costed chondrocytes and increased incorporation of [3H]TdR from (73 +/- 9) Bq (control group) to (272 +/- 55), (327 +/- 38), and (415 +/- 32) Bq, respectively (P < 0.01). The activity of VAP-A was weaker than that of VAP-B, and VAP-C had no activity. VATP 10 and 20 mg.kg-1 by local injection into the cross-section fracture area accelerated healing of radial fracture. The healing rate of VATP-treated group was higher (75%) than that of control group (25%) (P < 0.05). CONCLUSION: VATP accelerated fracture healing by stimulating proliferation of chondrocytes and osteoblast precursors.

A natural vaccine candidate strain against cholera.

E1 Tor Vibrio cholerae (EVC) strains may be classified into two kinds-epidemigenic (EEVC) strains and non-epidemigenic (NEEVC) strains-based on a phage-biotyping system. A large number of EEVC strains have been screened for toxigenic and putative colonization attributes. One such naturally occurring strains (designated IEM101) has been found which is devoid of genes encoding cholera toxin (CT), accessory cholera enterotoxin (ACE), zonula occludens toxin (ZOT), but possesses RS1 sequences and toxin-coregulated pilus A gene (icpA) although icpA is poorly expressed. It expresses type B pili but does not possess type C pili. It is an E1 Tor Ogawa strain and does not cause fluid accumulation in rabbit ilcal loop tests. Active immunization of rabbits with strain IEM101 elicited good protection against challenge with virulent strains of V. cholerae O1. Oral administration caused no side effects in 15 human volunteers, colonized the gut for four to ten days and elicited good immune responses.

Dammarane saponins from Panax ginseng.

From the dried roots of Panax ginseng two new minor dammarane saponins named koryoginsenoside-R1 and -R2 were isolated, along with fourteen known saponins. On the basis of spectral and chemical evidence, the structure of the new saponins were elucidated as 6-O-[trans butenoyl-(1-->6)-beta-D-glucopyranosyl]-20-O-beta-D- glucopyranosyl dammar-24-en-3 beta,6 alpha,12 beta,20(S)-tetrol and 3-O-[beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl]-20-O-[beta-D- glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] dammar-22-en-3 beta,12 beta, 20(S),- 25-tetrol, respectively.

[Experimental study on the effect of abstinence with herbal preparation composite dong yuan gao].

The effect of herbal preparation "Composite Dong Yuan Gao" (CDYG) on the animal model of drug dependence was studied. After the morphinistic models of white rats and mice were made by ever increasing doses of morphine each time, the animals were divided into large dose CDYG group, small dose CDYG group, sustained morphine group, and control group. The withdrawal symptoms were observed after naloxone was given to the animals intraperitoneally. The results showed that in the CDYG groups withdrawal symptoms of the addicted white rats were alleviated, the number of jumps of the addicted mice was reduced, the scores of the principal symptoms of the animals were decreased, while the body weights of the animals were gained. Comparing with the control group, the difference was significant (P < 0.01). The effects of the CDYG enhanced in the large dose group, which suggests that the CDYG has abstinence effects.

[Identification of fruits of Xanthium sibiricum Patr. and X. mongolicum Kitag].

The protein electrophorogram, tissue separation picture and complete powder characteristic picture of these two kinds of fruit of siberian cocklebur are made for the first time, and TLC chromatograms of different solvent extracts are also presented. All these present a scientific basis for crude drugs selling, breed identification and composition analysis of the fruit of siberian cocklebur.

[Studies on the hypoglycemic effect of Rhodiola sachalinensis A. Bor. polysaccharides].

Rhodiola sachalinensis polysaccharides (RSP) isolated from the root of Rhodiola sachalinensis was demonstrated to decrease the levels of blood sugar, liver glycogen and total blood lipid when injected peritoneally to mice at 50, 100 and 200 mg/kg once or for 7 successive days. In addition, RSP was found to decrease blood sugar and total blood lipid in various experimental hyperglycemias induced by injection of adrenaline, glucose and alloxan in mice. RSP exhibit inhibiting effect on the glycogen enhancement induced by glucose as well as strenghtening effect on the glycogen decomposition induced by adrenaline and alloxan.