AIM: To screen the lentiviral vector carrying siRNA with higher efficiency of suppressing the sphingosine-1-phosphate receptor 2 (S1P2) gene expression in the primarily cultured corpus cavernosum smooth muscle cells of spontaneously hypertensive rats (SHR).METHODS: SHR and SD rats (n=5 each) were used for primarily culturing corpus cavernosum smooth muscle cells.The cells were randomly divided into 6 groups: SHR siRNA-1, SHR siRNA-2, SHR siRNA-3, SHR GFP, SHR control (SHR non-transfection group), and SD control (SD rat control group).Each group had 5 samples with 1.0×105 cells of each sample.At 72 h after transfection (MOI=60) with lentiviral vectors carrying S1P2 siRNA into the SHR corpus cavernosum smooth muscle cells, the expression of GFP was observed under fluorescence microscope.The protein expression of S1P2, ROCK1, ROCK2 and eNOS in the corpus cavernosum smooth muscle cells, and the mRNA expression of S1P2, ROCK1 and ROCK2 were determined by by Western blot and RT-PCR.RESULTS: The transfection efficiency of the corpus cavernosum smooth muscle cells in SHR siRNA-1, SHR siRNA-2, SHR siRNA-3 and SHR GFP groups were>80%.Compared with SHR control group, the mRNA levels and the protein expression of S1P2, ROCK1 and ROCK2 in SHR GFP group showed no remarkable changes, while those in SHR siRNA-1, SHR siRNA-2, SHR siRNA-3 and SD control groups were significantly lower than those in SHR control group (P<0.05).The protein expression of eNOS in SHR siRNA-1, SHR siRNA-2, SHR siRNA-3 and SHR GFP groups were not significantly changed as compared with SHR control group, but that in SD control group was significantly higher than that in SHR control group.CONCLUSION: Three groups of siRNA lentiviral vectors targeting S1P2 inhibit the expression of S1P2 in the corpus cavernosum smooth muscle cells of SHR, and by silencing the S1P2 expression, the expression of ROCK1 and ROCK2 is inhibited.Among them, siRNA-1 has the highest inhibitory efficiency.

AIMTo investigate the effects of protein tyrosine kinase on the inflammation and airway remodeling in lung of guinea pigs with bronchial asthma.

METHODS30 adult male guinea pigs were randomly divided into 3 groups (n=3): control group (C group), asthmatic group(A group)and genistein group (B group). Asthmatic model was established by ovalbumin intraperitoneal injection and ovalbumin inhalation. The total cell and the proportion of inflammatory cell in bronchial alveolar lavage fluid(BALF), inflammatory cell infiltration and index of remodeling of bronchiole were measured, respectively. The expression of p-tyrosine in lung tissue was examined by immunohistochemistry.

RESULTSThe total cell and proportion of eosinophil in BALF of A group were significantly higher than that of C group (P < 0.01), but compared with A group, the total cell and proportion of eosinophil in BALF of B group were much lower (P < 0.01). The number of eosinophile and lymphocyte of bronchiole in A group were significantly higher than that of C group (P < 0.01), but compared with A group, the number of eosinophile and lymphocyte in bronchiole of B group were much lower (P < 0.01). Compared with A group, the remodeling of bronchiole of B group was significantly relieved (P <0.01), there was no difference between B and C group (P > 0.05). Immunohistochemistry indicated that in A group the p-tyrosine was more positively expressed at the bronchial smooth muscle, bronchial epithelium, smooth muscle of vessel and inflammatory cell, especially at smooth muscle of bronchi and vessel and inflammatory cell than that of C group (P <0.01), there was no difference between B group and C group (P > 0.05).

CONCLUSIONPTK played a key role in inflammation and bronchial remodeling in lung of guinea pigs with bronchial asthma. The Protein tyrosine kinase inhibitor genistein could prevent and inhibit the inflammation and bronchial remodeling in lung of guinea pigs with bronchial asthma.

Airway Remodeling ; physiology ; Animals ; Asthma ; physiopathology ; prevention & control ; Genistein ; pharmacology ; Guinea Pigs ; Inflammation ; prevention & control ; Male ; Ovalbumin ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; physiology ; Random Allocation

Aneurysmal subarachnoid hemorrhage accounted for 70% of subarachnoid hemorrhage. The high fatality and disability rate affected patients′ quality of life. Aneurysmal subarachnoid hemorrhage should be considered as a chronic cerebrovascular disease, and it is very important to take early intervention and strengthen the secondary prevention to the patients with risk factors for disease. This paper reviewed the research status on quality of life of patients with aneurysmal subarachnoid hemorrhage and its influencing factors, in order to provide a reference for neurological physicians.

Objective To explore the effect of low constant direct current (LCDC) in the medullary cavity on repairing of rabbit radial defect as to evaluate its clinical value. Methods DBX was put into the segmental osteoperiosteum defect of rabbit bilateral radius. The rabbits were divided into 2 groups: group A (subject group) with the defects stimulated by LCDC and group B (control group) with the defects left untreated. In week 2,5 and 9, 4 animals from each group were put into death, and the radius was taken for study using photograph, electron microscope, histological & morphologic technique, and the contents of the Calcium (Ca) and ALP were determined. Results Compared with group B, the healing of group A was better. The contents of Ca and ALP increased obviously in week 2 (P

Objectives:To study the value of fructose-1,6-diphosphate-supplemented parenteral nutrition in elder abdominal surgical patients.Methods:thirty-two patients were randomly allocated into study group and control group,receiving isocaloric(83.6 kJ/kg?d~(-1)) and isonitrogenous(0.2 g/kg?d~(-1)) PN from post-operation day 1 to post-operation day 6.The study group received 10g of FDP every day and did not receive Glycophos.Plasma albumin,pre-albumin and phosphate lever were measured and EKG was examined before surgery and days 1 and 7 after surgery.The level of 3-methylhistidine(3-MH) in urine was measured and nitrogen balances were calculated postoperatively on day 1,3,5 and 7.Results:The blood phosphate levels were normal in two groups before and after surgery.Compared with preoperation,plasma albumin and pre-albumin levels were both decreased in two groups and pre-albumin level was markedly decreased in the control group(P

Objective To investigate the experience of treating Budd-Chiari syndrome through orthotopic liver transplantation.Methods The clinical data of LTx performed on 9 patients with Budd-Chiari syndrome from December 2003 to April 2010 were retrospectively analyzed. We summarize the preoperative image and surgical experience,and observe the occurrence of postoperative complications and survival. Results Budd-Chiari syndrome was diagnosed in 9 patients by the preoperative abdominal CT enhancement and vascular reconstruction,and cavity venography was done to observe obstruction and sub-type of CAVA vein.All 9 patients were subjected to cadaveric liver transplantation.Eight cases accepted classic non bypass type,and one accepted living related right lobe liver transplantation. Postoperative triple immunosuppressive regimen included tacrolimus,mycophenolate mofetil,and hormone.The average follow-up periods for all these 9 patients were 32.8 months (13 to 61 months). One patient died from the tumor recurrence at 35th month after the operation.Two patients received re-transplantation for the lost of the graft.One recipient received the donor liver with medium steatosis,and the re-transplantation was performed on the12th day after the first transplantation due to the primary non function of the graft.The other one received the secondary liver transplantation at 6th month after the first transplantation due to the biliary complication and died from the liver tumor recurrence. Among all the 9 cases,seizure disorder (1 case),dysfunction of duodenal papillary muscle (1 case) and small-for-size syndrome (one case) occurred after the operation.Pulmonary infection occurred in 4 cases:3 cases due to the bacterial infection and 1 due to the fungal infection. Neither outflow obstruction nor the recurrence of the Budd-Chiari syndrome occurred in this study.The 1- and 2-year survival rate after the operation was both 100％,and 3-year survival rate post-transplantation was 88.9％ (8/9).Conclusion Liver transplantation can be the ideal treatment to the Budd-Chiari syndrome based on the definite clinical diagnosis,accurate imaging evaluation and eligible modus operandi.

Objective To explore a new method of functional reconstruction of hand digits and joints with free transfer of foot tissues so as to increase the success rate of the operation.Methods After micro-anatomic study of the plantar and dorsal metatarsal arteries,retrograde and free grafts of foot tissues pedicled with plantar metatarsal arteries were designed and applied in transplantation to treat 76 cases of hand digital or joint defects.The surgeries included 58 cases of transfer of the second toe,four cases of transfer of composite tissues of the second toe, eight cases of transfer of proximal interphalangeal joint,and six cases of nail flap transfer.Results The mi- cro-anatomic study found that the first plantar metatarsal artery was anatomically constant and the diameter of its branch to the second toe was larger than that of the first dorsal metatarsal artery.Flaps survived in 75 of the 76 patients(98.7%),with fine appearance and significantly improved function.One patient who had received free transfer of the second toe to reconstruct the thumb function had to undergo a second repair with infraclavicula skin tube because of refractory arteriospasm of anastomosed vessels.Conclusion Transfer with free retrograde grafts of foot tissues pedicled with plantar metatarsal artery to reconstruct hand functions can effectively improve the success rate of the operation,because it is free of the shortcomings of great anatomic variation of blood vessels and time-consuming and complex procedures in conventional transfer.

OBJECTIVETo investigate the levels of secretions from the prostate and seminal vesicles and their association with the expressions of aquaporins (AQP) in the prostatic tissue and seminal vesicles of castrated rats.

METHODSWe randomly divided 18 eight-week-old male SD rats into a control, a castration, and a testosterone (T) replacement group. Four weeks after surgical castration, we detected the plasma T level and measured the volumes of the secretions and the expressions of AQPs 3, 7, and 10 - 12 in the prostate and seminal vesicles of the rats.

RESULTSThe plasma T level was significantly lower in the castrated models ([30. 98 ± 28. 84] ng/dl) than in the rats of the control ([700.78 ± 123.8] ng/dl) and T replacement groups ([688.08 ± 132. 47] ng/dl) (P <0. 05). The castration group, in comparison with the control and T replacement groups, showed remarkably reduced ratios of prostatic secretion volume / prostate weight ([11.1 ± 0.30] vs [2.32 ± 0.61] and [2.13 ± 0.56] %, P <0. 05) and seminal vesicle secretion volume / seminal vesicle weight ( [4. 78 ± 1. 97 ] vs [57. 36 ± 11. 86] and [55. 74 ± 7. 21] %, P < 0. 05). Immunohistochemistry revealed the expressions of AQPs 3 and 7 in the epithelial envelop and cytoplasm and that of AQP 11 the in endothelial envelop and cytoplasm of the prostate and seminal vesicles. Western blot exhibited significantly lower expressions of AQPs 3, 7, and 10 - 12 in the prostate and seminal vesicles of the castrated rats than in the animals of the control and T replacement groups (P <0. 05).

CONCLUSIONSignificant decreases of the secretions from the prostate and seminal vesicles may be related to the reduced expressions of AQPs 3, 7, and 10 - 12 in the prostatic tissue and seminal vesicles in castrated rats.

Animals ; Aquaporins ; metabolism ; Humans ; Male ; Orchiectomy ; Prostate ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Seminal Vesicles ; metabolism ; Testosterone ; blood

Holmium ; Humans ; Kidney ; Lithotripsy, Laser ; Shock, Hemorrhagic ; Ureteroscopy

Objective To detect the expressions of TLR7/9, IRF7 and IRAK1 in condyloma accuminatum (CA) lesions and to investigate their possible roles in pathogenesis of CA.Methods The expressions of TLR7/9, IRF7 and IRAK1 in 23 cases of CA lesions were tested by immunohistochemistry. Results TLR9-positive cells were observed both in the epidermis and in the papillary layer of dermis.The density and positive rate of TLR9-positive cells in the papillary layer of CA lesions were significantly higher than those in their normal borderline tissues ( P <0.05 ) .TLR7-positive cells, IRF7-positive cells and IRAK1-positive cells were observed mostly in the papillary layer of dermis.There were no obvious difference of their density and positive rate between the CA lesions and their normal borderline tissues ( P>0.05 ) . Conclusions The TLR9 expression was up-regulated,while the TLR7, IRF7 and IRAK1 expression were un-changed in CA lesions.Our results suggested that the downstream of TLR7/9 signaling pathways in CA could not be provoked despite the up-regulation of TLR9 expression, which lead to the difficulty to HPV removal and the recurrence of CA.