Objective To investigate the gene silencing,apoptosis induction and the suppression of proliferation in vivo transfected by UTMD techniques associated with shRNA techniques. Methods The survivin-shRNA expression vector was constructed. Nude mice were randomly arranged into 3 groups:control group, plasmid injection and ultrasound (P + US), P + UTMD group. Histological examination were evaluated. Protein expressions of Survivin and proliferating cell nuclear antigen (PCNA), Bcl-2, Bax,Caspase-3, Ki-67, nucleostemin (NS), p53 were investigated by immunohistochemistry. Results In transplanted tumors experiment, comparing with those in C and P + US groups, protein expressions of PCNA,Ki-67,Bcl-2, Survivin, NS were down-regulated markedly, while those of Bax, Caspase-3 and P53 were up-regulated significantly ( P ＜ 0.05). Conclusions UTMD combined with shRNA technique can induce apoptosis and inhibit proliferation significantly, without causing any apparently adverse effect,representing a new,promising technology that can be used in the tumor gene therapy and research.

Objective:To transfect genes using ultrasound targeted microbubble destruction (UTMD) techniques and observe the effects of RNA interference on cervical cancer (HeLa) cell line in silencing survivin gene and inducing apoptosis. Methods: Recombinant expression plasmid of short hairpin RNA (shRNA) targeting survivin gene was constructed. It was co-treated with microbubbles and transfected to cultured HeLa cells followed by exposure to ultrasound (P+UTMD group). Moreover, blank control group (C), plasmid group (P), ultrasound exposure group (US), plasmid and ultrasound exposure group (P+US), plasmid+ Lipofectamine group (P+L) were used as controls, respectively. Transfection efficacy was evaluated by observing the red fluorescence in the cells by fluorescent microscopy and flow cytometry(FCM). Ultrasound intensity and exposure time were optimized. Cell apoptosis was investigated using flow cytometry analysis, Hoechst staining, and DNA ladder method. Expression of survivin mRNA was assessed by RT-PCR. Results: Restrictive enzyme digestion and sequencing analysis verified that the recombinant plasmid was successfully constructed. UTMD significantly increased gene transfection efficacy in cultured HeLa cells (P<0.01). Gene transfer was the most prominent at ultrasound intensity of 1.0 W/cm2 and exposure time of 3 min (P<0.01). RT-PCR showed that the expression of survivin mRNA in P+UTMD group was inhibited by (83.33±2.73)%. The differences were significant compared with any other groups (P<0.01). FCM analysis showed that the apoptosis ratio in P+UTMD group was significantly increased as compared with other groups (P<0.01). Hoechst staining and DNA ladder showed that apparent apoptosis and DNA ladder were detected only in P+UTMD and P+L groups. Conclusions:UTMD effectively enhances the transfection efficacy of expression plasmid. It is a novel and effective non-viral gene transfer system and has promising foreground. UTMD mediates RNA interference silenced survivin gene and induces significant cell apoptosis, which provides a new method for tumor research and gene therapy.

Objective To investigate the effects of Ruangan granule on transforming growth factor-β1(TGF-β1)/Smads signaling pathway in liver fibrosis in rats. Methods A total of 105 Wistar rats were randomly divided into normal control group, model group and colchicine, Dahuang-Zhechong pill group, high-, medium- and low-dose Ruangan granule groups (n=15 in each group). Liver fibrosis was induced by carbon tetrachloride and a high-cholesterol diet. After modeling, the low-, medium- and high-dose Ruangan granule groups were intragastric administrated Ruangan granule mixed suspension 3.6, 7.2, 14.4 g/(kg?d), respectively;Dahuang-Zhechong pill group was administrated with Dahuang-Zhechong pellets mixed suspension of 0.18 g/(kg?d);the colchicine group was intragastric administrated with colchicine mixed suspension of 0.108 mg/(kg?d);and the normal control group and the model group were intragastric administrated with the equal volume of distilled water. All rats were intragastric administrated for 8 weeks. The expressions of TGF-β1, Smad3 and Smad7 proteins in the liver tissue were detected with immunohistochemical staining method. The expressions of TGF-β1, Smad3, Smad7 mRNAs in the liver tussue were detected by RT-PCR. Results The expressions of TGF-β1 (2.59 ± 0.99 vs. 0.43 ± 0.21) and Smad3 (2.56 ± 0.67 vs. 0.41 ± 0.18) proteins and TGF-β1 mRNA (2.25 ± 0.21 vs. 0.71 ± 0.09) and Smad3 (2.34 ± 0.03 vs. 0.78 ± 0.12) mRNAs in the model group were significantly increased than those in the normal control group (all P<0.01). Compared with the model group, the expressions of TGF-β1 (1.12 ± 0.27 vs. 2.59 ± 0.99) and Smad3 (1.05 ± 0.34 vs. 2.56 ± 0.67) proteins in the high-dose Ruangan granule group decreased significantly, the expression of Smad7 increased significantly (2.33 ± 0.62 vs. 0.36 ± 0.18), and the expressions of TGF-β1 (1.09 ± 0.11 vs. 2.25 ± 0.21) and Smad3 (1.10 ± 0.02 vs. 2.34 ± 0.03) mRNAs decreased significantly, the expression of smad7 mRNA (1.18 ± 0.13 vs. 0.38 ± 0.11) increased significantly (P<0.05). Conclusions Ruangan granule can regulate the TGF-β1/Smads signaling pathway via down-regulation of TGF-β1, Smad3 and up-regulation of Smad7 in liver fibrosis in rats.

Objective To observe theRuangan granule on liver fibrosis in rats liver pathology change, the influence of hepatic function and hepatic fibrosis indexes, and to discusses the mechanism of its action to provide the basis for clinical prevention and treatment of liver fibrosis.Methods A total of 105 Wistar rats were randomly divided into a normal control group, a model group and a colchicines group, and Dahuang-Zhechong pill group, high-, medium- and low-doseRuangan granule groups (n=15 in each group). Liver fibrosis was induced by carbon tetrachloride and a high-cholesterol diet. After modeling, the low-, medium- and high-doseRuangan granule groups were intragastric administratedRuangan granule mixed suspension 3.6, 7.2, 14.4 g/(kg?d), respectively;Dahuang-Zhechong pill group was administrated with Dahuang-Zhechong pellets mixed suspension of 0.18 g/(kg?d); the colchicine group was intragastric administrated with colchicine mixed suspension of 0.108 mg/(kg?d); and the normal control group and the model group were intragastric administrated with the equal volume of distilled water. All rats were intragastric administrated for 8 weeks. HE staining and Masson trichromatic collagen staining were used to observe the pathological changes of liver While the change of AST, ALT, PH, TP and serum HA, LN, C-Ⅳ, PCⅢin blood serum were detected. Results Masson trichromatic collagen staining showed that, the percentage of liver collagen fiber area in rats of theRuangan granule high-dose group was significantly decreased (7.06 ± 1.18) % compared with model group (23.49 ± 1.34) %, colchicine group (11.35 ± 1.83) %, rhubarb worm pill group (15.27 ± 1.22) %,Ruangan granule medium-dose group (14.52 ± 1.75) %, and low dose group (16.08 ± 1.56) % (P<0.05 orP<0.01). Compared with model group,Ruangan granule high-dose group rats serum AST (75.86 ± 5.23 U/Lvs. 157.62 ± 24.04) U/L, the ALT (80.15 ± 5.94 U/Lvs. 160.58 ± 26.47) U/L, PH (52.58 ± 4.98μg/Lvs. 98.66 ± 6.75)μg/L significantly reduced, TP (74.19 ± 3.56 g/Lvs. 51.73 ± 5.92)g/L increased significantly (P<0.01).Ruangan granule high-dose group rats serum HA (277.22 ± 106.34 ng/mlvs. 553.19 ± 172.38 ng/ml), LN (89.82 ± 5.68 ng/mlvs. 134.25 ± 10.64 ng/ml), C-Ⅳ (47.94 ± 8.65 ng/mlvs. 84.18 ± 13.83 ng/ml), PCⅢ (16.53 ± 4.88 ng/mlvs.31.57 ± 5.35 ng/ml) decreased significantly in the model group (P<0.01).ConclusionRuangan granule has obvious effects for resisting liver fibrosis.

OBJECTIVETo observe the curative effects of biliary drainage via endoscope (ENBD) combined with Chinese drug medication on acute biliary tract infection (ABTI) and the influence of treatment on complement 3 (C3), C-reactive protein (CRP) and interleukin-6 (IL-6).

METHODSSixteen patients with ABTI were randomly assigned to two groups: the 9 patients in the combined treatment group (CTG) treated with ENBD combined with Chinese medicine and the 7 patients in routine treatment group (RTG) treated with ENBD alone. Another 18 patients with simple gallbladder stone were taken as the control group (CG). The curative effect was observed and the serum concentrations of C3, CRP and IL-6 were determined before and after treatment.

RESULTSBefore treatment, the concentrations of CRP and IL-6 were significantly higher and C3 lower in all ABTI patients than those in patients with simple gallbladder stone (P < 0.05 or P < 0.01). After treatment, the general condition of patients was improved, the recovery time of intestinal tract function was shortened and the concentrations of C3, CRP and IL-6 significantly decreased in CTG, with the effects better than those in RTG respectively (P < 0.01 or P < 0.05).

CONCLUSIONENBD combined with Chinese drug medication shows favorable curative effects on ABTI. Treatment with Chinese medicine according to syndrome differentiation could decrease blood level of pro-inflammatory cytokines and promote recovery of the injured immune function.

Adult ; Aged ; C-Reactive Protein ; metabolism ; Cholangitis ; blood ; therapy ; Combined Modality Therapy ; Complement C3 ; metabolism ; Drainage ; instrumentation ; methods ; Drugs, Chinese Herbal ; therapeutic use ; Endoscopy, Digestive System ; Female ; Humans ; Interleukin-6 ; blood ; Male ; Middle Aged ; Phytotherapy ; Treatment Outcome

Objective To optimize the volatile oil extraction and inclusion process of Wenweiyang capsules. Methods An orthogonal test was adopted in this study. The extraction technology was optimized for the yield of volatile oil regarding the amount of water loaded, grain size of medicinal material, and decoction time as factors. The inclusion technology was optimized for the inclusion yield and volatile oil inclusion rate using the ratio ofβ-CD:oil, amount of water and grinding time as factors. Results The optimized extraction parameters were as follows:breaking medicinal material through 10 mesh screen, adding 6 fold volume of water and extracting for 5 h. The optimized inclusion progress was grinding at theβ-CD:oil ratio of 81, loading equivalent amount of water and grinding for 30 minutes. The average yield of volatile oil is 1. 72%, the average inclusion rate is 93. 01% and the average volatile oil inclusion rate is 74. 82%. Conclusion The extraction and inclusion technology is simple, reliable, which can effectively retain the volatile oil and provide evidence for the preparation of Wenweiyang capsules.

Objective To explore the predictive value of neutrophil gelatinase associated lipocalin (NGAL) for contrast-induced acute kidney injury (CI-AKI) after emergency percutaneous coronary interventions (PCI) in patients with relative normal renal function.Methods A total of 73 patients with relative normal renal function undergoing PCI were enrolled in this prospective multicenter clinical study.Serum NGAL was measured by point-of-care test.Receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of NGAL for CI-AKI.Results CI-AKI occurred in 5 patients (6.85％).The concentration of NGAL was higher in the CI-AKI group than in the non-CI-AKI group.ROC curve indicated that the area under the receiver operating characteristic was 0.755,0.761 and 0.809,respectively.Conclusions Serum NGAL at baseline,4 h and 8 h after procedure are served as a good biomarker for early diagnosis of CI-AKI after PCI.Therefore,NGAL might become an early and quick marker for CI-AKI in the future.

Objective To explore and integrate the key techniques used in the surveillance and forecast of schistosomiasis in the water regions along the Yangtze River,so as to provide technical support for identifying rapidly the risk of schistosomiasis transmission and implementing control measures targeting the risk. Methods According to the distribution of water systems and water regions along the Yangtze River in Jiangsu Province,the demonstration sites for surveillance and forecast of schistoso? miasis were set across the province,where the integration and demonstration of the techniques regarding monitoring of Schistoso?ma japonicum infection in sentinel mice,human and animal activities,release of forecast information,and emergency treat?ment of water regions at risk of infection were performed. The pattern of human and animal activities was compared with the S. ja?ponicum infection in sentinel mice in the demonstration sites,and the operability of the release of information and emergency treatment of the risk of S. japonicum infection was evaluated. Results A total of 50 demonstration sites for surveillance and forecast of schistosomiasis were set in fixed anchor points,opening of the navigation lock to the Yangtze River,freight terminal, agritainment places,ferry,large construction places,and places for guaranteeing the Youth Olympic Games in 23 counties(dis?tricts)of 5 cities,Jiangsu Province. During the period between May and September,2014,the infectivity of water body was monitored by using 5 batches of sentinel mice,with a 99.06%(4 954/5 001)gross recovery rate of mice. S. japonicum infection was detected in a demonstration site,and an infected mouse was found,with a 0.02%(1/4 933)gross positive rate of sentinel mice. The field survey showed 2 088 person?times contacting water,including 91.95%(1 920/2 088)contacting water due to the production such as capturing fish,harvesting and cultivating crops,and repairing and building boats,and 8.05%(168/2 088)contacting water due to the life activity,such as fishing,washing vegetables and playing with water. The people contacted water predominantly in August and September(49.57%). A total of 859 boats containing 1 877 boatmen were observed,68.22%(586/859)of which were fishing boats containing 1 306 fishermen(69.58%). There were 32 sheep found in 4 demonstration sites,and 3 sheep were seen in the demonstration site with infected sentinel mouse. Four blue forecasts(emergence of water con?tact)and one orange forecast(S. japonicum?infected sentinel mouse detected)were released across the province,with one fore?cast map released which showed 5 sites with fishing and one site with sheep grazing,one emergency response system initiated, mollusciciding implemented in 10 hm2 high?risk regions,120 sheep grazed in fence,and 35 fishermen given health?education materials,schistosomiasis examination and preventive therapy. In addition,no acute schistosomiasis occurred in the demonstra?tion site with S. japonicum?infected sentinel mice. Conclusions The integration and demonstration of the techniques regarding monitoring of S. japonicum infection in sentinel mice,human and animal activities,release of forecast information,and emer?gency treatment of water regions at risk of infection provides an effective approach for the large?scale surveillance and forecast of schistosomiasis.

BACKGROUNDRetinopathy of prematurity (ROP) has become one of the leading causes of visual loss in children. Vascular endothelial growth factor A (VEGF-A) is the principal stimulator of angiogenesis. VEGF was differentially spliced from exon 8 to exons 8a and 8b to form two families: the pro-angiogenic VEGFxxx family and the anti-angiogenic VEGFxxxb family. Previous research has shown variable effeteness of bevacizumab in inhibiting retinal neovascularization in ROP. This study aimed to investigate whether the effectiveness of this inhibition depends on the relative ratio of the two VEGF isoforms.

METHODSIntravitreal bevacizumab injection (IVB) was performed in the oxygen-induced-retinopathy (OIR) mice on postnatal day 12 (P12) (intravitreal phosphate buffered saline (PBS) injection as control). The Evans blue perfused retina were used to test the retinal neovascularization-leakage (NVL) area and non-perfusion (NP) area.

RESULTSThe retinal NVL and NP area in the IVB group were significantly smaller than the intravitreal PBS injection group (IVP group). On P17, the protein level of total VEGF isoforms was significantly inhibited compared to IVP group (P < 0.05) while VEGF(165)b isoform was slight reduced (P > 0.05). The switch from pro-angiogenic isoforms to anti-angiogenic isoforms after IVB could be found. The relative protein expression of VEGF(165)b isoform was significantly higher in IVB group than in IVP group (P < 0.05) on P17 which was correlated with the reduced ischemia-induced angiogenesis in OIR mice after IVB.

CONCLUSIONSThe anti-angiogenic effectiveness might depend on the relative high expression of VEGF(165)b after intravitreal bevacizumab injection. Anti-angiogenic therapy is a more effective therapy for ROP.

Angiogenesis Inhibitors ; administration & dosage ; Animals ; Animals, Newborn ; Antibodies, Monoclonal, Humanized ; administration & dosage ; Bevacizumab ; Disease Models, Animal ; Intravitreal Injections ; Mice ; Mice, Inbred C57BL ; Protein Isoforms ; analysis ; Retinal Neovascularization ; prevention & control ; Retinopathy of Prematurity ; drug therapy ; Vascular Endothelial Growth Factor A ; analysis

Objective To establish the surveillance and risk assessment system of schistosomiasis in Jiangsu Province,so as to provide technical support for timely understanding of the risk of schistosomiasis transmission and implementation of target-ed control measures. Methods The surveillance sites of schistosomiasis were assigned according to the epidemic status and en-demic type of schistosomiasis as well as the characteristics of the water system,and the prevalence of Schistosoma japonicum in humans and domestic animals,and snail status were investigated. In addition,the quality control of serum detection of S. japon-icum infections was performed. The prevalence of human and animal S. japonicum infections,snail status and missing diagnosis of serum detection were analyzed and compared among regions. Results A total of 27 surveillance sites of schistosomiasis were set up in 26 counties of 10 cities,Jiangsu Province,including 14 sites in transmission-interrupted villages and 13 sites in trans-mission-controlled villages,and 15 sites in marshland and lake regions,9 sites in plain regions with water network and 3 sites in mountainous region. In the 27 surveillance sites,a total of 16 617 residents were screened for S. japonicum infection by using dipstick dye immunoassay(DDIA),and 326 were sero-positive,with a sero-prevalence of 1.96%(2.17%for men and 1.8%for women). Of the 326 individuals undergoing parasitological examination,2 positive cases were detected in the marshland and lake region,with a S. japonicum human prevalence of 0.01%. Of the 762 floating population detected,10 were positive for blood test,with a sero-prevalence of 1.31%,and no egg-positive individuals were detected. No infection was found in the 476 do-mestic animals. Of the 746 settings surveyed,a total of 240.7 hm2 snail area was detected,with a mean snail density of 0.06 snails/0.1 m2,and no infected snails were found. There were 780 quality-control sera detected in 26 surveillance sites of schisto-somiasis,and the gross coincidence rate was 95.13%,with misdiagnosis rate of 1.28%and missing diagnosis rate of 19.23%. Conclusion The surveillance sites of schistosomiasis show reasonable distribution in Jiangsu Province,and the endemic situa-tion of schistosomiasis appears a low level in the whole province.