Objective To comparatively investigate the characteristics of bone metabolism in forelimb malformation WHBE rabbits based on the OPG/RANK/RANKL system.Methods Healthy male 2-2.5-month old WHBE rabbit ( HWR) , Healthy Japanese rabbit ( HJR) and forelimb malformation WHBE rabbit ( FMWR) were used in this study and divided into 3 groups, HWR, HJR and FMWR groups, with 10 rabbits in each group.The shape of forelimb and mean gray value observed from X-ray film were examined, and the bone tissue micro-morphology was analyzed using HE staining.Re-al-time fluorescence quantitative PCR was used to determine the expression of osteoprotegerin ( OPG) and receptor activator of NF-KB ligand ( RANKL) mRNA.The expressions of OPG, receptor activator of NF-KB ( RANK) and RANKL protein in serum and bone tissue were assayed by enzyme-linked immunoassay ( ELISA) and immunohistochemistry, respectively. Results Compared with the HWR and HJR groups, rabbits in the FMWR group appeared abnormal in toutuous forelimb and thinner bone cortex.The mean gray values of X-ray in the FMWR group were lower than that in the HWR group ( P<0.05).There were significant differences between the FMWR and healthy rabbits (HWR and HJR) in the following pa-rameters:RANKL mRNA expression and RANKL/OPG mRNA ratio in the liver(P<0.05, P<0.01), serum protein expression of RANK and RANKL and RANKL/OPG ratio (P<0.05, P<0.01), and positive index of expression of RANKL protein and RANKL/OPG ratio in the bone tissue(P<0.05, P<0.01).Furthermore, the gene expression lev-els of OPG and RANKL of HWR were significantly higher than that of HJR(P<0.05, P<0.01).Conclusions The FMWR show some abnormal symptoms in bone metabolism as well as decrease of bone quality and histological changes of bone micro-structure, due to the significant increase of RANKL/OPG ratio of FMWR.The breed differences between WHBE and Japanese rabbits in RANKL mRNA expression level may be one of factors inducing limb malformation in the WHBE rabbits.

Objective To investigate the association between variable number of tandem repeat(VNTR) polymorphism of endothelial nitric oxide synthase(eNOS) gene and cerebral infarction(CI).Methods The genotypes of 152 patients with CI were detected by polymerase chain reaction(PCR) and compared with control group. Multiple regression analysis was performed to assess the independent roles of the polymorphism of eNOS and other risk factors.Results The ab genotype distribution frequency of eNOS gene in CI group(22.36%) was higher than that in control group(12.28%), and a allele frequency of eNOS gene in CI group(12.5%) was also higher than that in control group(7.0%). Both differences between CI group and control group were significant(all P

In 34 hypothalamic slices of rats, spontaneous discharging of 63 paraventricular neurons was extracellulariy recorded by glass microelectrode. When the slices were perfused with artificial cerebrospinal fluid (ACSF) containing noradrenaline (NA) (10-6 mol/L), firing rates of 20 units significantly increased, those of 8 units decreased or even ceased, and those of 35 units, not affected. When low Ca2+ high Mg2+-ACSF was applied to block synaptic transmission, of 20 units which were excited by NA, 14 units still had excitatory responses and 6 units had no significant response to NA. When synaptic transmission was blocked, of 8 units which were inhibited by NA, 7 units still had inhibitory responses and only one unit had no significant response to NA. The results of this experiment strongly suggest the existence of direct effects of NA on paraventricular neurons.

Objective:To evaluate the meaning and value of high-frequency ultrasound in the diagnosis of carpal tunnel syndrome (CTS).Methods:In this study,48 patients (unilateral hand)with CTS were analyzed.The thickness of transverse carpal ligaments at the pisiform bone was measured using high-fre-quency ultrasound.Open carpal tunnel release procedure was performed in the 48 CTS patients,and the thickness of transverse carpal ligaments at the hamate hook bone measured using vernier caliper under di-rect vision.The accuracy of thickness of transverse carpal ligaments was evaluated using high-frequency ultrasound.High-frequency ultrasound measurement of thickness of transverse carpal ligaments at the ha-mate hook bone and pisiform bone,and determination of the diagnostic threshold measurement index using receiver operating characteristic (ROC)curve,sensitivity and specificity were performed and the correlation between the thickness of transverse carpal ligaments and nerve conduction study (NCS)ana-lyzed.Results:The thickness of transverse carpal ligaments in the CTS patients were (0.42 ±0.08)cm (high-frequency ultrasound)and (0.41 ±0.06)cm (operation)at hamate hook bone,and there was no significant difference between the two ways (t =0.672,P>0.05 ).The optimal cut-off value of the transverse carpal ligaments at hamate hook bone was 0.385 cm,the sensitivity 0.775,and the specificity 0.788.The optimal cut-off value of the transverse carpal ligaments at the pisiform bone was 0.315 cm, the sensitivity 0.950,and the specificity 1 .000.The transverse carpal ligaments thickness and wrist-in-
dex finger sensory nerve conduction velocity (SCV),wrist-middle finger SCV showed a negative correla-tion.Conclusion:High frequency ultrasound measurements of thickness of transverse carpal ligaments is a valuable method for the diagnosis of CTS.

Objective To investigate the Laparoscopic management for Nutcraker Syndrome(NCS) with resection of fibrous ring and placing extravascular stent.Methods This was a retrospective analysis of clinical data and treatment process of cases from March 2010 to February 2015 in urology department,affiliated hospital of Guizhou medical university.Five cases with NCS,4 males and 1 female;age were 28-40 years,mean age was 35 years,all cases were afflicted with gross hematuria and flank pain,the history of gross hematuria were 6-72 months.3 cases were afflicted with proteinuria.Duplex ultrasound scanning before surgery revealed the compressed left renal vein (LRV) between the aorta and the superior mesenteric artery(SMA),with peak velocity 110-132 cm/s,an average of 121 cm/s.The flow velocity of LRV in the renal hilum were 18-25 cm/s,an average of 21 cm/s.CT scanning showed that the stricture segment diameter of LRV were 1.2-2.5 mm,an average of 1.8 mm;and the max diameter of proximal dilatation of the LRV in renal hilum were 8.3-15.2 mm,an average of 10.1 mm.The ratio between the dilated segment inner diameter and the stricture segment were 3.4-9.5.Bleeding from the left ureteral orifice was detected by cystoscopy in 3 cases.5 cases were treated by resection of fibrous ring and placing extravascular stent with Laparoscopic management,and the average length of extravascular stent was 4.0 cm.Results The operation was successful in the 5 cases.The average operation time was 83 min.The average blood loss was 65 ml.Hematuria gradually reduce 5-6 days and resolved 7-20 days after surgery in 5 patients.Proteinuria was disappeared successful 2 weeks after surgery in 3 patients.There was no recurrence at 8-24 months' follow-up.3 days after surgery Doppler ultrasound showed the stricture segment diameter of LRV were 3.8-5.6 mm,an average of 4.9 mm;the ratio between the dilated segment inner diameter and the stricture segment decreased were 1.1-2.0,an average of 1.6;the peak velocity of compressed LRV were 25-45 cm/s,an average of 34 cm/s.6 months after surgery,CTA result showed no LRV compression in the aortomesenteric region;the max diameter of LRV in renal hilum were 7.9-9.8 mm and 6.0-8.8 mm in the aortomesenteric region of LRV.Conclusion Etiology of NCS exist a fibrous ring around the left renal vein outflow of the inferior vena cava besides the commonly anatomic extrinsic compression on the LRV as it crosses between the superior mesenteric artery and the aorta.The Laparoscopic management for NCS with resection of fibrous ring and placing extravascular stent is an effective minimally invasive treatment.

Objective To explore the effect of heating temperature and heating time on the dissolution characteristics of phenolic acids in water extract of Danshen Decoction. Methods The contents of tanshinol, protocatechuic aldehyde and salvianolic acid B in water extract of Danshen Decoction were determined by reverse-phase high performance liquid chromatography (RP-HPLC) . The changes of the contents were monitored by classic constant temperature acceleration method. Results Of the three components, the content of salvianolic acid was decreased with the increase of heating temperature and the prolongation of heating time, and the degradation characteristics of the salvianolic acid B met the first-order kinetics equation. Conclusion The water-soluble components have less stability in the water extracts of Danshen Decoction. Lignum Santali Albi and Fructus Amomi accelerate the degradation speed of Radix Salviae Miltiorrhizae, suggesting that we should pay attention to the influence of heating temperature and heating time on the content of salvianolic acid B in the extraction and concentration process.

Objective To evaluate the efficacy of combination therapy with M and α receptors antagonist for the treatment of double-J stent related lower urinary tract symptoms.Methods From May 2013 to May 2014,131 cases,including 71 male and 60 female cases,were accepted the doubte-J stent indwelling after the ureteral lithotripsy,laparoscopic ureterlithotomy and pyeloureteroplasty.Their data was retrospectively reviewed.The age ranged from 29 to 64 years old,mean (47.4 ± 15.2) years old.They were divided into 4 groups randomly,including group A (control group,n =30),no drugs were taken;group B (tamsulosin group,n =34),0.2 mg tamsulosin was taken qd;group C (solifenacin group,n =32),5 mg solifenacin was taken qd;group D (tamsulosin combined with solifenacin group,n =35),0.2 mg tamsulosin and 5mg solifenacin were taken qd.The IPSS scores,QOL scores and visual analogue pain scale (VAPS) scores were assessed pre-operation,1 week after operation,and 4 weeks after operation,respectively.Results All patients were followed-up until the end of this study.In each time point,the IPSS scores in group A was 9.01 ± 2.79,13.18 ± 3.79 and 13.79 ± 3.76,respectively.In group B,the IPSS scores were 7.89 ± 4.29,12.39 ±3.90 and 12.21 ±3.87,respectively.In group C,the IPSS scores were 7.94 ±4.27,12.70 ±4.01 and 11.98 ±4.69,respectively.In group D,the IPSS scores were 8.21 ±3.18,11.97 ±5.03 and 8.17 ± 3.25,respectively.Significant difference in total IPSS scores and obstruction symptom scores were shown between pre-and post-operation (P ＜ 0.05).Comparing to other groups,group D exhibited the significant improvement in IPSS scores 1 and 4 weeks after the operation (P ＜0.05).4 weeks after operation,the QOL scores in group D was significantly lower than that in other groups (P ＜ 0.05).While the VAPS scores didn t show significant differences among those groups (P ＞ 0.05).Conclusion M and α receptors antagonist combination therapy can significantly improve lower urinary tract symptom due to indwelling double J stents.

OBJECTIVE: To observe the effects of recipes for replenishing qi and activating blood on p16, p21, proliferating cell nuclear antigen (PCNA), cyclin D1 and cyclin E gene expressions in the liver of aging rats. METHODS: A recipe for replenishing qi and a recipe for activating blood were administered to aging rats respectively, and the effects of the above recipes on the expressions of senescence related genes (p16, p21, PCNA, cyclin D1 and cyclin E) were examined by RT-PCR and Western blotting methods. RESULTS: The expressions of p16, p21 and cyclin D1 mRNAs and proteins in the liver of the untreated aging rats were up-regulated, while the expressions of PCNA and cyclin E mRNAs and proteins decreased. As compared with the untreated aging rats, both recipes could down-regulate the expressions of cyclin D1 mRNA and protein and up-regulate the expressions of cyclin E mRNA and protein, but had no obvious effects on the expressions of mRNAs and proteins of p16, p21 and PCNA. CONCLUSION: Recipes for replenishing qi and activating blood can improve the liver cell proliferation of aging rats via down-regulating the expressions of cyclin D1 mRNA and protein and up-regulating the expressions of cyclin E mRNA and protein.

The curcumin prodrugs, which could be selectively activated in tumor cells, were prepared to establish a basis for the targeted chemotherapy for cancer. On the basis of the molecular structure of curcumin, the N-maleoyl-L-valine-curcumin (NVC), N-maleoyl- glycine-curcumin (NGC) were chemically synthesized and identified by IR and NMR spectroscopy. After treatment with these two prodrugs for 6 - 24 h, the rates of growth inhibition on human bladder cancer EJ cells and renal tubular epithelial (HKC) cells were detected by MTT colorimetry. Our results showed that after the treatment with 20 micromol/L - 40 micromol/L NVC and NGC for 6 - 24 h, the growth inhibitory effects on EJ cells were 6.71% - 65.13% (P < 0.05), 10.96% - 73.01% (P < 0.05), respectively, in both dose- and time-dependent manners. When compared with the curcumin of same concentrations, the growth inhibitory effects of these two prodrugs on HKC cells were significantly decreased (P < 0.01). It is concluded that activation of curcumin prodrugs via hydrolysis functions of cellular esterase could inhibit the growth activities of tumor cells, and reduce the side effects on normal diploid cells. This provided a novel strategy for further exploration of tumor-targeted chemotherapeutic drugs.
Antineoplastic Agents, Phytogenic/*pharmacology ; Curcumin/*pharmacology ; Dose-Response Relationship, Drug ; Prodrugs/*chemical synthesis ; Prodrugs/*pharmacology ; Tumor Cells, Cultured ; Urinary Bladder Neoplasms/*pathology

OBJECTIVETo compare the effects of TCM therapeutic principles of tonifying Shen (TS), benefiting Qi (BQ), invigorating Pi (IP) and activating blood circulation (ABC) herbs in regulating the gene expression in senescence related cell cycle.

METHODSDrug sera containing TCM herbs of the above-mentioned principles were used to treat the aged human diploid fibroblast cell line 2BS. The effect of TCM on the senescence related cell cycle and its related gene expression (P16INK4, Cyclin D1 and PCNA) were examined by means of cell proliferative doublings, flow cytometry, RT-PCR and Western blot analysis.

RESULTSTCM herbs of TS and BQ could improve the cell cycle, down-regulate the P16 and Cyclin D1 mRNA/protein expression, up-regulate PCNA mRNA/protein expression, while TCM herbs of IP and ABC showed insignificant effect on these indexes.

CONCLUSIONTCM herbs of TS and BQ have effect in improving cell cycle, it may be achieved through promoting the P16 pathway of gene expression.

Aging ; genetics ; Animals ; Cell Cycle ; drug effects ; Cells, Cultured ; Cellular Senescence ; genetics ; Cyclin D1 ; biosynthesis ; genetics ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; genetics ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; cytology ; Gene Expression Regulation ; Lung ; cytology ; Male ; Medicine, Chinese Traditional ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley