OBJECTIVETo investigate the effects of serum containing Gumibao (Chinese character: see text) on the proliferation and differentiation of osteoblast induced by dexamethasone.

METHODSOsteoblasts were extracted from skulls in newly born (within 24 hours) SD rats, and digested with collagenase. The first passage of cells were used for experiments. Cells were cultured in the medium containing different concentrations of dexamethasone (0, 10(-8), 10(-7), 10(-6), 10(-5) ,10(-4) mol/L). Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks. Accordingly, following the treatment of 10(-5) mol/L dexamethasone for 1 week, cells were cultured in the medium with serum containing Gumibao (Chinese character: see text). One week after Cumibao (Chinese character: see text) treatment, cells were stained with Alkaline phosphatase and collagen I and PCNA were examined by Western-blot. However, the observation of numbers of mineralized nodes with alizarin red stain required one more week.

RESULTSHigh concentration of dexamethasone could inhibit the expression of PCNA, collagen I, alkaline phosphatase and reduce the number of mineralized nodes of osteoblast, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

CONCLUSIONHigh concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; drug effects ; physiology ; Proliferating Cell Nuclear Antigen ; analysis ; Rats ; Rats, Sprague-Dawley

Background Neovascular glaucoma (NVG) is a serious ocular disease which may cause blindness.The primary pathogenesis of NVG is ischemic retinopathy derived by central retinal vein occlusion (CRVO) and diabetic retinopathy (DR).Clinical characteristics of NVG are variable based on the difference of primary diseases,such as CRVO and DR.However,there is a few studies regarding the diffcrcnces of NVG initiated by CRVO and DR.Objective This study was to compare the clinical characteristics in NVG patients secondary to CRVO and DR.Methods A series case observational study was carried out in Hiserve Hospital of Qingdao University from January 2009 to June 2012.Twenty-nine eyes of 27 patients with NVG caused by CRVO (10 eyes of 10 patients) and DR (19 eyes of 17 patients) were included.The history of underlying diseases,course of NVG,intraocular pressure(IOP),fundus findings and complications after treatment were analyzed and compared between the CRVO-derived NVG and DR-derived NVG.All patients underwent panretinal photocoagulation,improving microcirculation therapy,anti-glaucoma (drug or surgery) and causative disease treatment,and some of them received vitrectomy or/and cataract surgery.Two eyes from each group received intravitreal injection of ranibizumab.The follow-up time in both groups was (14.00±10.13) months and (17.89±12.52) months,respectively.Results The median time of underlying disease was 3.3 months (2 weeks to 6 months) in the CRVO patients and 11.1 months (4 to 36 mouths) in the DR patients,with a significant difference between them (Z =-2.40,P＜0.05).CRVO-derived NVG progress was much faster than that of DR-derived NVG.The number of the eyes with visual acuity improvement after treatment was 2 in the CRVO-derived NVG and 15 in the DR-derived NVG;while the number of the eyes with unchanged or worse visual acuity was 8 and 4 in the CRVO-derived NVG eyes and the DR-derived NVG eyes (x2 =9.38,P＜0.01).The difference of IOP in pre-and post-treatment was (37.00±9.91)mmHg in the CRVOderived NVG eyes and (8.92±12.05)mmHg in the DR-derived NVG eyes,showing a significant difference between them (t =6.30,P＜0.01).In the CRVO-derived NVG eyes,optic disc edema,retinal hemorrhage,and vein dilatation were seen in 6 eyes,and mild optic disc edema and retinal hemorrhage were observed in 4 eyes.After treatment,fundus could not be seen in 4 eyes,in other 2 eyes optic disc and retinal laser spots were unclearly observed.In addition,pale optic disc and retinal vessel occlusion appeared in 2 eyes,and silver wire-like arteries exhibited in 2 eyes.In pre-treated DR-derived NVG eyes,fundus could not be seen in 8 eyes and Ⅲ-Ⅳv stages of DR findings appeared in 11 eyes.After treatment,retinopathy was stabilized in 16 eyes of 15 cases.Advanced retinopathy(V-Ⅵ stages of DR findings) was revealed in 3 eyes of 3 cases.The incidence of the complication after treatment was 100.0％ in the CRVO-derived NVG eyes and 21.1％ in the DR-derived NVG eyes (x2=5.18,P＜0.05).Conclusions The clinical characteristics of NVG secondary to CRVO and DR are variable,an appropriate treatment option should be selected according to different features of NVG.

In early 2011, the serious outbreak of porcine pseudorabies virus (PRV) infection suddenly recurred in Henan and neighboring Provinces. To investigate the etiology of massive infection with PRV, 16 800 serum samples, 905 porcine epidemic diarrhea virus (PEDV) back-feeding tissues, and 56 PR gene deleted live vaccines were colleted from January 2011 to May 2013 to detect PRV field infection using a PRV gE antibody test kit. The gE and TK genes of 11 new epidemic PRV strains were sequenced by PCR, and their molecular characteristics were analyzed. Moreover, virus titer determination, protective test against PRV, and vaccine potency testing were performed. The results showed that the detection rate of PRV field infection-positive pig farms was 68.06%, and the overall positive rate of PRV field infection in serum was 38.47%; the positive rates in breeding sows, breeding boars, reserve pigs, and commercial pigs were 40.12%, 30.88%, 54.67%, and 26.52%, respectively. The new epidemic strains were in the same evolutionary branch and belonged to the virulent strain group. Compared with the classical PRV strain, the virulence of new epidemic strains changed a little. The length of gE gene was 1 787 bp, and the length of TK gene was 963 bp. The nucleotide homologies of gE and TK genes to Chinese reference strains were 98.2%-99.8% and 98.90%-99.6%, respectively, and the amino acid homologies were 97.1%-99.8% and 97.5%-99.4%, respectively. Commercial vaccine had a 100% protective effect against the new epidemic strains. The positive rate of PRV field infection was 0% in vaccine and 40.44% in back-feeding tissues. The results confirmed that PRV field infection rates were rising sharply among pigs in Henan and neighboring Provinces after 2011. The main virulence genes of new epidemic PRV strains did not change significantly over the years. PR gene deleted live vaccines had no PRV field infection and could completely resist the attack of new strains. The virus carriage of breeding boars and reserve pigs and the serious PRV field infection in PEDV back-feeding tissues were the main causative factors for massive infection with PRV and epidemic outbreak in Henan and neighboring Provinces from 2011 to 2013.
Amino Acid Sequence ; Animal Feed ; analysis ; virology ; Animals ; China ; epidemiology ; Epidemics ; Female ; Herpesvirus 1, Suid ; chemistry ; classification ; genetics ; isolation & purification ; Male ; Molecular Sequence Data ; Phylogeny ; Pseudorabies ; epidemiology ; virology ; Sequence Alignment ; Sequence Homology, Amino Acid ; Sus scrofa ; Swine ; Swine Diseases ; epidemiology ; virology ; Viral Proteins ; chemistry ; genetics

OBJECTIVETo compared the activity and yield rate of osteoblast obtained by different collagenase digestion methods, to find a better way to extract osteoblast for the experimental researches of osteoporosis.

METHODSTen 24-hour-old SD rats were were euthanized. The cranium of rats were removed and cuted into blocks of 1 mm x 1 mm size. After digested by trypsin for 15 min, all the cranium were divided into two equal parts, and randomly divided into two groups which would be digested by type I collagenase and type II collagenase separately for two times. The rat cells of the two groups were cultured in thermostat incubator with 5% CO2 under the condition of 37 degrees C. The primary culture osteoblasts were counted by using a haemacytometer after digestion and 72 hours later. The second generation osteoblasts cultured 48 h were dyed by NBT/BCIP staining solution, and were detected by quantitative measurement with PNPP.

RESULTSThe cells had irregular shapes. The results of cell counting showed that the cell number of type I group was larger than type 11 group. Alkaline phosphatase dyeing were positive. Detecting of alkaline phosphatase using the method of PNPP showed that the absorbance value in type I group were higher than type II group (P<0.05).

CONCLUSIONTwo types of collagenase are both suitable for the in vitro culture of rat osteoblasts. The activity and yield rate of osteoblasts in type I group are higher which could provide more stable seed cells for the treatment of osteoporosis.

Animals ; Cell Count ; Cell Culture Techniques ; methods ; Collagenases ; metabolism ; Female ; Male ; Osteoblasts ; cytology ; Rats ; Rats, Sprague-Dawley

Small cell carcinoma of the esophagus (SCCE) is a rare and aggressive malignant tumor with a poor prognosis. The optimal disease staging system and treatment approaches have not yet been defined. This study aimed to evaluate the prediction of different staging systems for prognosis and treatment options of SCCE. We retrospectively accessed the clinicopathologic characteristics, treatment strategy, and prognosis of 76 patients diagnosed with primary SCCE between 2001 and 2011. The 1-, 2-, 3-, and 5-year overall survival rates were 58%, 31%, 19%, and 13%, respectively. Univariate analysis showed that the 2002 American Joint Committee on Cancer (AJCC) tumor-node-metastasis (TNM) classification (P = 0.002), Veterans Administration Lung Study Group (VALSG) stage (P = 0.001), predisposing factors (P < 0.001), T category (P = 0.023), and M category (P < 0.001) were prognostic factors for overall survival. Multivariate analysis showed that the 2002 AJCC TNM stage (P < 0.001) was the only independent prognostic factor for survival. The value of the area under the receiver operator characteristic (ROC) curve (AUC) of the 2002 AJCC TNM staging system was larger than that of VALSG staging system with regard to predicting overall survival (0.774 vs. 0.620). None of the single treatment regimens showed any benefit for survival by Cox regression analysis. Thus, the 2002 AJCC TMN staging system improved the prediction of SCCE prognosis; however, the optimal treatment regimen for SCCE remains unclear.
Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Small Cell ; classification ; pathology ; therapy ; Cisplatin ; administration & dosage ; Combined Modality Therapy ; Esophageal Neoplasms ; classification ; pathology ; therapy ; Esophagectomy ; methods ; Etoposide ; administration & dosage ; Female ; Humans ; Lymph Node Excision ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; methods ; Paclitaxel ; administration & dosage ; Radiotherapy, High-Energy ; Retrospective Studies ; Societies, Medical ; Survival Rate ; United States

OBJECTIVETo investigate the effects of osthole on proliferation of neonatal rat osteoblast and the mechanism.

METHODSTen 24 hours old SD rats were executed by dislocating. The cranium of rats were removed and cut into blocks of 1 mm x 1 mm size. After digested by trypsin for 15 min, the cranium were digested by type I collagenase for one hour two times. The mixed cells were cultured in thermostat incubator with 5% CO2 under the condition of 37 degrees C. To identify the cells, ALP staining and alizarin red staining were performed after cultured 48 h and 28 d. The osteoblasts were randomly divided into five groups. Cells were treated with osthole at concentrations of 100, 50, 25, 12.5, 0 micromol/L. CCK-8 method was used to evaluate the proliferation after 24 h,48 h and 72 h. The expression of PCNA and beta-catenin protein were detected through the method of Western Blot after one week.

RESULTSThe cells had irregular shapes and showed typical features of osteoblast. The results of ALP staining and alizarin red staining were both positive. CCK-8 detection showed that the osthole with final concentration of 100 micromol/L inhibited the proliferation of osteoblast after 24 h, while the osthole with final concentrations of 50 micromol/L and 25 micromol/L displayed the inhibition effect after 48 h. The osthole of 12.5 micromol/L had no obvious influence on the proliferation of osteoblast. The result of Western Blot showed that osthole reduced the expression of PCNA and beta-catenin protein in a dose-dependent manner.

CONCLUSIONThe osthole with final concentrations of 100, 50, 25 micromol/L inhibited the proliferation of osteoblast (P < 0.05). The osthole with final concentrations of 12.5 micromol/L had no obvious influence on the proliferation of osteoblast (P > 0.05). These findings demonstrate that osthole may inhibit the proliferation of osteoblast by regulating the Wnt/beta-catenin signaling in osteoblast.

Animals ; Animals, Newborn ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Coumarins ; pharmacology ; Female ; Male ; Osteoblasts ; cytology ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Sincalide ; metabolism ; beta Catenin ; metabolism

OBJECTIVETo explore the effects of percutaneous transhepatic radiofrequency ablation (PRFA) combined with tumor edge of percutaneous absolute ethanol injection (PEI) on liver cancer adjacent to major blood vessels.

METHODSSeventy five patients with liver cancer adjacent to major blood vessels were randomly divided into two groups: PRFA+PEI therapy group (38 cases) and PRFA control group (37 cases). Tumor necrosis rate, AFP levels, local recurrence rate, median for survival time and cum survival were used as the evaluation index to evaluate the efficacies of the two methods.

RESULTSTumor necrosis rates of the therapy group and the control group were 84.2% and 54.1% (P < 0.01), respectively; AFP levels of therapy group and control group at 1, 3, 6 and 12 months after treatment were (105.0 ± 35.5) μg/L, (28.4 ± 4.3) μg/L, (58.6 ± 6.7) μg/L, (89.5 ± 12.5) μg/L and (137.2 ± 34.6) μg/L, (84.2 ± 18.4) μg/L, (106.6 ± 20.3) μg/L, (173.7 ± 32.0) μg/L, respectively. The rates of therapy group was significantly lower than of control group. Local recurrence rates of the therapy group and control group were 2.6%, 7.9%, 13.2% and 31.6% vs 10.8%, 21.6% , 40.5% and 62.1% (P < 0.05) at 3, 6, 12 and 24 months after treatment, respectively. Median for survival time of the therapy group and control group were 28.0 ± 2.8 months and 19.0 ± 3.6 months, respectively. Cum survival of the therapy group and control group were 84.2%, 78.9%, 60.5% and 31.6% vs 78.4%, 67.6%, 37.8% and 8.1% (P < 0.05) at 6, 12, 24 and 36 months after treatment, respectively.

CONCLUSIONPEI as a supplementary treatment of PRFA can effectively improve the treatment of liver cancer adjacent to major blood vessels and significantly reduce the local recurrence rate and improve long-term survival rates.

Adult ; Aged ; Bile Duct Neoplasms ; Carcinoma, Hepatocellular ; pathology ; therapy ; Catheter Ablation ; Combined Modality Therapy ; Ethanol ; administration & dosage ; Female ; Humans ; Liver Neoplasms ; pathology ; therapy ; Male ; Middle Aged ; Retrospective Studies ; Survival Rate ; Treatment Outcome

OBJECTIVETo detect serve acute respiratory syndrome-associated coronavirus (SARS-CoV) and SARS-like-CoV in fruit bats captured in Guangzhou and its vicinity.

METHODSTotally 927 bats of 9 species (Cynopterus sphinx, Rousettus leschenaulti, Miniopterus schreibersi, Hipposideros pratti, Rhinolophusasinicus, Scotophilusakuhlii, Hipposideros Pomona, Rhinolophus affinis, and Rhinolophus pusillus) captured in Guangzhou and its vicinity from September 2004 to November 2005 were available for this investigation, from which 3,043 samples (813 throat swasb, 524 sera, 853 lung tissues and 853 colorectal tissue specimens) were obtained. SARS-Cov and SARS-like-CoV were detected in these specimens using diagnostic kit for novel coronavirus N protein (ELISA), SARS-CoV Virus RNA detection kit, fluorescence PCR, Genchip, RT-PCR and cell isolation culture methods.

RESULTS AND CONCLUSIONNo SARS-CoV and SARS-like-CoV were detected in the 3043 samples, indicating the current absence of SARS-CoV and SARS-like-CoV in the bats captured in Guangzhou and its vicinity.

Animals ; China ; epidemiology ; Chiroptera ; virology ; Disease Vectors ; Enzyme-Linked Immunosorbent Assay ; Humans ; Nucleocapsid Proteins ; metabolism ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; isolation & purification ; metabolism ; Severe Acute Respiratory Syndrome ; epidemiology ; transmission ; virology

OBJECTIVETo explore the specific efficacy of different moxibustion techniques in treatment of common diseases and clinical indications, and compare the specificity in clinical indications and efficacy among different moxibustion techniques so as to guide clinical practice better.

METHODSThe modern computerization and data mining technology were adopted to set up moxibustion literature database. The relevant literature of moxibustion techniques in recent 60 years were collected, screened, examined, extracted and analyzed statistically so as to explore the advantages of different moxibustion techniques in clinical treatment.

RESULTS(1) Of 2,516 literature, moxa stick, moxe cone and moxa device were used in the highest frequency in internal medicine department, for 730 times, 278 times and 102 times respectively. The warm needling technique was used in the highest frequency, for 70 times in the surgical department. (2) In the dermatology department, the curative rate with moxa cone was the highest, 75%. In the ear-nose-throat department, the warm needing technique and moxa device achieved the highest curative rate, 49% for both of them. In the internal medicine department and surgical department, the curative rate of warm needling technique was 53% and 58% respectively. In the gynecology department, the curative rate of moxa device was the highest, 59%. In the pediatrics department, the curative rate of moxa cone was the highest, 80%. (3) The numbers of priority disorders, frequency ≥20 times: 24 kinds of disease for moxa stick, five kinds of disease for moxa cone, 2 kinds of disease for warm needling technqiue and one disorder for moxa device. Facial paralysis, diarrhea, lumbar and leg pain and elbow and knee swelling pain were of the highest priority, treated with these 4 moxibustion techniques, with a certain of literature research values. (4) The warm needling technique achieved the better efficacy on elbow and knee swelling pain, lumbar and leg pain and diarrhea compared with the other three techniques and the curative rate was higher. The moxa device tecnique achieved the higher curative rate for facial paralysis compared with the other three techniques.

CONCLUSIONThrough the comparison of application frequency, curative rate, clinical application frequency in disorders and the efficacy of priority disorders in the treatment with different moxibustion techniques, it is found that moxa stick, moxa cone and moxa device are simple in manipulation, safe and effective. Hence, they can be extensively used in the treatment of common disorders in every department in clinic. The warm needling technique acts on the body by the co-work of needling and warming stimulation of mugwort. It achieves the particular effect on the disorders with complicated etiologies compared with the other three techniques. It can be chosen in priority for the disorders caused by blockage in meridian and collateral and stagnation of qi and blood.

Clinical Trials as Topic ; Data Mining ; Humans ; Meridians ; Moxibustion ; methods ; Treatment Outcome

Objective Apoptosis contributes to the instability of the atheresclerotie (AS) lesions.The vulnerable plaque was identified in vivo by detecting the apoptosis with radiolabeled annexin Ⅴ in an atherosclerotic rabbit model Methods Eight male New Zealand white rabbits on 2% cholesterol diet for 2 weeks had abdominal aortic balloon injury and fed a 2% cholesterol diet for another 15 weeks (AS group),3 rabbits fed a normal rabbit chow for 17 weeks without balloon injury served as contrels.Annexin Ⅴ labeled with 99Tcm was then intravenously administered and planar whole-body images were captured using a gamma camera in the left lateral position.The entire length of the abdominal aorta was explanted for ex vivo imaging with gamma camera.The aorta then was divided into several segments according to the severity of AS.The segments were separated weighted and counted in an gamma counter for the absorptive dose of annexin per gram of tissue.Histology examinations were made on specimens.Results At 2 hours post annexin Ⅴ injection,clear delineation of radiolabel within the abdominal aorta could be evidenced in vivo gamma.imaging.After explanation of the aorta,ex vivo imaging showed a robust uptake of radiotracer in the infradiaphragmatic aorta corresponding to the in vivo images and conforming to the macroscopic distribution of atherosclerotic lesions.The uptake of radiolabel was absent in areas without grossly visible atheroeclerotic lesions.The in vivo and ex vivo images identified plaque areas were identical and corresponded histological results on the explanted specimen.The aortic specimen was divided into 18 segments on lesions.The magnrity of the lesions (14/18) manifested as type Ⅳ or type Ⅴ lesions of AHA classification (vulnerable lesions),except segmentsl-4,which manifested as type Ⅰ or type Ⅱ lesions.The thickness of fibrous cap (TFC) and the ratio of cap and lipid nuclear (RCN) were significantly reversely correlated to the unit radioactivity counts,and the correlation between RCN and the unit radioactivity counts was more significant than that between TFC and the unit radioactivity counts (r=-0.904,P＜0.01,and r=-0.8,P＜0.01).Apoptosis detection (TUNEL):annexin Ⅴ intake in plaques was positively correlated to apoptotic index(r=0.651,P=0.012).Conclusion Noninvasive Annexin Ⅴ imaging could be used to detect vulnerable atherosclerotic plaques in vivo.