OBJECTIVETo screen the lentiviral vector carrying siRNA and capable of significantly suppressing the ROCK2 gene expression in the corpus cavernosum smooth muscle cells of spontaneously hypertensive rats (SHR).

METHODSWe designed and synthesized 4 siRNA fragments targeting the ROCK2 gene and packaged them into lentiviral vectors. We collected corpus cavernosum smooth muscle cell samples from 5 male SHRs and randomly divided them into groups A (non-transfection control), B (GFP lentiviral transfection), C, D, E, and F (lentiviral transfection with siRNA fragments 1 -4 targeting the ROCK2 gene). Each group consisted of 5 samples and each sample 3 x 10(4) cells. At 48 hours after transfecting MOI = 80 into the SHR corpus cavernosum smooth muscle cells, we detected the expression of GFP under the fluorescent microscope and the mRNA expression of the ROCK2 gene by RT-PCR.

RESULTSThe transfection efficiency of the SHR corpus cavernosum smooth muscle cells was > 50%. Compared with group A, the expression of ROCK2 mRNA in the corpus cavernosum smooth muscle cells showed no remarkable change in group B (P > 0.05) but was inhibited very significantly in C ([43.91 +/- 8.19]%), D ([47.15 +/- 6.64]%), and F ([25.7 +/- 6.03]%) (P < 0.01), and significantly in E ([16.81 +/- 5.94]%, P < 0.05).

CONCLUSIONWe successfully constructed 4 lentiviral vectors carrying siRNA targeting the ROCK2 gene, all of which can significantly suppress the ROCK2 expression in the SHR corpus cavernosum smooth muscle cells, and one has a highly strong inhibitory effect.

Animals ; Gene Silencing ; Genetic Vectors ; Lentivirus ; genetics ; Male ; Myocytes, Smooth Muscle ; Penis ; cytology ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; Random Allocation ; Rats ; Rats, Inbred SHR ; Transfection ; methods ; rho-Associated Kinases ; genetics

OBJECTIVE:To compare the quality of domperidone tablets produced by 3 different factories METHODS:The quality of different products was investigated according to the standards of Ministry of Public Health and manufacture factories,including dissolubility,uniformity of dosage units,related substances and seals of blister packaging RESULTS:In 3 batches of products manufactured by Y,W and L,the contents of domperidone were 98 23%～99 74%,96 92%～98 31% and 97 42%～98 72%,and content uniformity A+1 8S was 2 11～3 38,3 97～5 25 and 5 49～7 47 respectively,which were in keeping with the Chinese pharmacopoeia;impurity R061668 was not found,and contents of impurity R052211 were 0 004%～0 024%,0 029%～0 072% and 0 003%～0 056%;the total impurity substances were 0 030%～0 095%,0 102%～0 128% and 0 232%～0 489%;the dissolubity(45min) was 95 09%～95 77%,91 25%～96 43% and 95 48%～96 58% respectively In experimental condition,no leakage was found from blister package of Y and W products,however,the leak rate of L product reached more than 38% CONCLUSION:According to this survey,the quality of product Y is the best,W is better

Objective To evaluate short-term fowllow-up results of castable ceramic-IPS Empress Ⅱ onlays in restoring severe tooth defects with low occlusogingival dimension from clinical aspects.Methods 41 units of the onlay were fabricated and applied in 37 patients,who were divided into experimental group and control group.The experimental group were restored with IPS EmpressⅡcastable ceramic onlay which was bonded by a dual-cure adhesive system.The control group were restored with mental onlay which was bonded by glass-ionomer cements.All these cases were followed at 6 and 12 months after cementation of the restorations.Metewands were established to evaluate its clinical effects.Results Onlay made from IPS Empress Ⅱ castable ceramic demonstrated natural and esthetical appearance.In addition,proximal contact,retention and marginal fit of the restorations were good.No gingivitis was found.Conclusion The reliability of onlay made from IPS Empress Ⅱ castable ceramic is greatly improved due to the strengthening effects of the lithium disilicate structures.In addition,the better esthetics compared to resin filling technique makes it a comparatively ideal all-ceramic system for onlays.

ObjectiveTo assess the feasibility of seeding adipose-derived stem cells (ADSCs) onto bladder acellular matrix grafts (BAMGs) for bladder reconstruction in a rabbit model.MethodsAutologous ADSCs were isolated,expanded and identified by flow cytometry.In the experimental group,ADSCs were seeded onto BAMGS for reconstructing bladder defects in 12 male rabbits.Unseeded BAMGs were used for bladder reconstruction in the control group of 12 rabbits.Cystography was performed at 24 weeks after grafts implantation.Following cystography,the animals were scarified and grafts were harvested； H&E and immunohistochemical staining were performed with cytokeratin AE1/AE3,smooth muscle α-actin and S-100 markers.ResultsFlow cytometry demonstrated that the ADSCs expressed CD90,CD44,CD105,CD166 and CD34,but not CD45 or CD106.The cells demonstrated good biocompatibility with BAMGs.At 24 weeks,in the experimental group,the reconstructed bladders reached a mean volume of (94.68 ± 3.31 )％ of the precystectomy bladder capacity.Complete regeneration of smooth muscle and nerve tissue was evident.Regenerated SMCs,urothelium and nerve cells stained positively for α-smooth muscle actin,AE1/AE3 and S100.In the control group,the mean bladder volume was (69.33 ± 5.05 )％ of the pre-cystectomy volume.Histologically,the control group was characterized by multi-layered urothelium without evidence for organized muscle or nerve tissue.Conclusion The tissue engineering bladder constructed by ADSCs and BAMG can be used as an ideal biomaterial to replace and repair the bladder.

OBJECTIVETo explore the relationship of bone cement distribution and the puncture angle in the treatment of thoracolumbar compression fractures with unilateral percutaneous kyphoplasty (PKP).

METHODSThe clinical data of 37 patients with thoracolumbar osteoporotic compression fractures underwent PKP between January 2013 to March 2014 were retrospectively analyzed, all punctures were performed unilaterally. There were 6 males, aged from 65 to 78 years old with an average of (71.83 ± 6.15) years; and 31 females, aged from 57 to 89 years old with an average of (71.06 ± 7.89) years. Imaging data were analyzed and puncture angle and puncture point were measured before operation. According to the measured data, the puncture were performeds during the operation. Distribution area of bone cement were calculated by X-rays data after operation. The effect of bone cement distribution on suitable puncture angle was analyzed; VAS score was used to evaluate the clinical effects.

RESULTSThe puncture angle of thoracic vertebrae in T8-T12 was from 28° to 33° with an average 30.4°; and the puncture angle of lumbar vertebrae in L1-L5 was from 28° to 35° with an average of 31.3°. Postoperative X-rays showed the area ratios of bilateral bone cement was 0.97 ± 0.15. Bilateral diffuse area were basic equal. Postoperative VAS score decreased significantly (1.89 ± 1.29 vs 7.03 ± 1.42).

CONCLUSIONThrough measure imaging data before operation with PKP,the puncture point and entry point can be confirmed. According the measured data to puncture during operation, unilateral puncture can reach the distribution effect of the bilateral puncture in the treatment of thoracolumbar compression fractures.

Aged ; Aged, 80 and over ; Bone Cements ; Female ; Fractures, Compression ; surgery ; Humans ; Kyphoplasty ; methods ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Spinal Puncture ; methods ; Thoracic Vertebrae ; injuries ; surgery

OBJECTIVETo observe the clinical effect of electric stimulation of long-term retaining needle on analgesia after laparoscopic cholecystectomy (LC) and the impacts on the post-surgical flatus time.

METHODSUnder static absorptive composite general anesthesia, 90 cases of LC were randomized into three groups, 30 cases in each one. In the control group, the analgesia was not applied after LC. In the analgesia-pumper group, the patient controlled intravenous analgesia (PCIA) was used. In the needle-retaining group, the electric acupuncture stimulator was used. The needles were inserted transversely at Riyue (GB 24), Qichong (ST 30) and Yanglingquan (GB 34) and fixed with sterile sticker. Separately, in 8 h and 24 h after surgery, the electric acupuncture stimulation with disperse-dense wave, 2 Hz/100 Hz frequency was applied continuously for 30 min. Visual analogue scale (VAS), adverse reactions such as vomiting and nausea and the postoperative flatus time in 2, 4, 8, 12, 24 and 36 h after surgery were observed and recorded in the three groups.

RESULTSIn 2, 4, 8, 12 and 24 h after surgery, VAS scores in the needle-retaining group and the analgesia-pumper group were all lower than those in the control group (P < 0.05, P < 0.01). The analgesia effect at the above time points in the needle-retaining group was better than that in the analgesia-pumper group (all P < 0.05). There was not adverse reaction in the needle-retaining group. But there were 3 cases of somnolence, 6 cases of nausea and 3 cases of vomiting in the analgesia-pumper group, and 2 cases of nausea and 1 case of vomiting in the control group. The flatus time was quite earlier in the needle-retaining group as compared with the other two groups [(14.77 +/- 4.99) h vs (18.50 +/- 4.22) h, P < 0.01; (14.77 +/- 4.99) h vs (18.17 +/- 4.69) h, P < 0.05].

CONCLUSIONThe electric stimulation of long-term retaining needle is safe and effective in analgesia after LC. It avoids the adverse reactions of analgesics and promotes postoperative flatus.

Acupuncture Analgesia ; instrumentation ; Adult ; Aged ; Cholecystectomy, Laparoscopic ; adverse effects ; Electroacupuncture ; instrumentation ; Female ; Humans ; Male ; Middle Aged ; Pain Management ; Pain, Postoperative ; etiology ; therapy

Objective To investigate fracture resistance and fracture patterns of maxillary anterior endodontically treated teeth restored with two different shapes of glass fiber post systems.Methods Twenty-four human sound maxillary anterior teeth of similar size were collected,and randomly divided into 3 groups,8 teeth each.After endodontical treatment,they were given the following treatments:Group A:parallel glass fiber posts (coltene parapost fiber lux) and composite core;Group B:taper glass fiber posts (coltene parapost taper lux) and composite core; Group C (control group):intact endodontically treated teeth.Teeth in Groups A and B reserved 2 mm dentin ferrule.Then the teeth were prepared and restored with IPS e.max Press all ceramic crowns.All the teeth were embedded in acrylic resin 2 mm below the cementoenamel junction (CEJ),with the silicon impression material stimulating periodontal membrane.All the specimen were loaded under a universal testing machine,at the palatal junction of incisor third and middle third,with an angle of 135 degree to the longitudinal axis of the tooth until fracture occurred,at a cross-head speed of 1.0 mm/min.Fracture loads and patterns were recorded.Statistical analysis was performed using SPSS software.Results The mean fracture loads in the three groups were as follows:(468.8±142.5) N,(440.2±202.7) N,and (459.0±147.6) N,respectively.There was no significant difference in fracture loads among groups (P＞0.05).The incidence of unfavorable fracture in Group A was higher than Groups B and C.Conclusions There is no significant difference in fracture strength between parallel and taper fiber post groups.However,the group with parallel fiber posts demonstrates a higher risk of unfavorable fracture than the group with taper fiber posts,which indicates that taper fiber posts are more favorable to preserve the remaining tooth structure.

Objective To assess the value of multi-slice computed tomography angiography (MSCTA) in the preoperative detection of aberrant hepatic arteries in patients scheduled to undergo pancreaticoduodenectomy.Methods Patients with pancreatic and peri-ampullary tumors were preoperatively studied using contrast-enhanced abdominal CT angiography (CTA).The results on hepatic arterial anatomy were compared with those obtained from digital subtraction angiography (DSA) and on surgical findings.Results Eighty-one patients were included into this study.DSA was carried out in 29 patients to evaluate tumor resectability,and 66 patients received surgery.Anomalous hepatic arteries were detected in 17 (21％) patients on CTA.Thirteen (16.0％) patients had a single arterial variant,and 4 (4.9％) patients had two arterial variants.One patient each was seen in the Michels type Ⅳ,Ⅶ,and Ⅷ respectively,while 2 patients each were seen in the Michels type Ⅲ and Ⅴ respectively.Five patients were diagnosed as the Michels type Ⅵ,and four as the Michels type Ⅸ.One patient demonstrated a rare variant which was not included into the Michels classification.MSCTA had an accuracy of 100％,a sensitivity of 100％,and a specificity of 100％.Regarding the traceability scores of hepatic arterial segment,there were no statistically significant differences between MSCTA and DSA.Conclusions MSCTA is an effective imaging tool to assess arterial anatomical variation around the pancreatic head.It is non-invasive,and it provides valuable information on the peri-pancre atic vascular anatomy before pancreaticoduodenectomy.

Objective To investigate the proliferation and differentiation characteristics of fibroblast like synovial cells(FLS)in rheumatoid arthritis(RA)in vitro and the mechanism of the immunosuppressive effect of differentiation inducers, such as all trans retinoid acid(ATRA), ealcitriol [1,25(OH)_2D_3] and dexamethasone(DEX). Methods FLS of knee synovial tissues from RA patients were cultured and identified in vitro in the presence or absence of ATRA, 1,25(OH)_2D_3 and DEX respectively. Synoviocyte proliferation in RA were measured by MTT colorimetrie assay and the survival fraction(SF)of FLS was evaluated. Cell cycle of FLS was observed using fluorescence-activated cell sorting(FCS)method in RA patients. Results The identified synovial cells in patients with RA were FLS(Vimentin and Fibronectin expression was positive), and hadn't been transformed or differentiated to adipocytes and osteoblasts with the three inducers. The SF of all RA-FLS interfered by ATRA, 1,25(OH)_2D_3 and DEX was much lower than that without drugs vehicle group in RA-FLS(P

Objective To investigate the proliferative characteristics of fibroblast-like synovial cells (FLS)in osteoarthritis in vitro and the mechanism of the immunosuppressive effect of total glucusides of paeony(TGP).Methods FLS of OA and non-inflamed synovium(NS)were cultured and identified in vitro in the presence or absence of TGP.After incubation,the survival fraction(SF)of FLS was evaluated by MTI' and the TNF-?,IFN-?and bFGF level in cultured FLS supernatant was measured by ELISA.The expression of FLS c-los mRNA and cell cycle of OA-FLS was observed by RT-PCR and flow eytometry respectively at the same time.Results No statistical significant differences were noted between the OA and NS FLS in pro- liferating double time.High doses of TGP suppressed FLS-SF more evidently in OA patients than in NS(P0.05).Conclusion High dose TGP can inhibit OA-FLS proliferation,modulate cy- tokine secretion and c-fos expression in OA.This suggests that TGP has immunosuppressive effect on OA syn- ovitis,probably by preventing the synovial hypertrophy in OA.