Objective To investigate the effect of phenylephrine on the morphology and the expression level of Nampt mRNA in the irradiated osteoblasts of the rats,and to clarify the possible protective effects of phenylephrine on this injury.Methods The osteoblasts cultured by conventional tissue were randomly divided into control group, phenylephrine group,simple irradiation group,and phenylephrine+irradiation group.The cells in phenylephrine group were treated with 100 mmol· L-1 phenylephrine 30 min before irradiation.The cells in simple irradiation group were irradiated with 8 Gy X-ray.The osteoblasts were observed by microscope and were collected at the time points of 8,16,32 and 64 h after irradiation.The Nampt mRNA expression level in osteoblasts was examined by RT-PCR.Results There was no obviously morphological and number changes of osteoblasts in phenylephrine group compared with control group.The osteoblasts in simple irradiation group were atrophied,especially at 8 h;the number of osteoblasts in simple irradiation group was 0.82,0.37,0.24 and 0.21 times as control group respectively at 8,16,32 and 64 h. Atrophy of osteoblasts was alleviated in phenylephrine+irradiation group;at 8, 16,32 and 64 h,the number of osteoblasts was 0.91,0.83,0.72 and 0.75 times as control group,and was 1.09,2.24,3.00 and 3.60 times as simple irradiation group. The expression level of Nampt mRNA in the osteoblasts in simple irradiation group was reduced compared with control group (P < 0.01) at 8, 16, 32 and 64 h.There were significant differernces of the expression levels of Nampt mRNA in the osteoblasts between phenylephrine+irradiation group and control group (P < 0.01),and there were significant differernces between phenylephrine + irradiation group and simple irradiation group (P < 0.01 ) at the different time points. Conclusion Phenylephrine could reduce the atrophy of osteoblasts in the rats induced by irradiation and up-regulate the expression level of Nampt mRNA.Its protective effect on irradiated injury may be related to the up-regulation of the expression level of Nampt mRNA.

Objective: To observe the therapeutic effect of sinew-regulating bone-setting manipulations for knee osteoarthritis (KOA) model rabbits and its impacts on the chondrocyte apoptosis rate and the levels of interleukin (IL)-1β and nitric oxide (NO). Methods: According to the random number table method, 30 New Zealand white rabbits were divided into a normal group (n=9) and a modeling group (n=21). Rabbits in the modeling group were used to establish KOA models with the modified Hulth method. At the 8th week, three rabbits were sacrificed to verify the model and the remaining 18 rabbits were randomly divided into a model group (n=9) and an intervention group (n=9). Rabbits in the normal group and model group were bred routinely without any intervention. Rabbits in the intervention group were treated with the sinew-regulating bone-setting manipulations, 10 min/time, once every other day for a total of 20 times. The Lequesne MG knee function rating was used to evaluate the behavioral differences of the rabbits in each group. The Pelletier score was used to evaluate the general changes of the rabbits. The Mankin score was used to evaluate the pathology of knee cartilages. The enzyme-linked immunosorbent assay and nitrate reductase methods were used to determine the levels of IL-1β and NO in serum and synovial fluid of each group, respectively. In situ terminal deoxynucleotidyl transferase-mediated nick and labeling method was used to determine the apoptosis of chondrocytes in each group. Results: Compared with the normal group, the scores of Lequesne MG, Pelletier and Mankin, and the levels of IL-1β and NO in the model group were increased (P<0.05), which indirectly indicated the success of the model. Compared with the model group, the scores of Lequesne MG, Pelletier and Mankin, IL-1β and NO levels, and chondrocyte apoptosis rate of the intervention group were decreased, and the differences were statistically significant (P<0.05). Conclusion: The sinew-regulating bone-setting manipulations can reduce the levels of IL-1β, NO, and chondrocyte apoptosis rate, and delay the articular cartilage degeneration, therefore, having a good therapeutic effect on KOA.

OBJECTIVETo investigate the mechanism of lemon peel essential oil (LPE) on the cariogenicity of Streptococcus sobrinus (Ss).

METHODSLPE was extracted by the authors, and the minimum inhibition concentration (MIC) was measured by disc diffusion method. The LPE was used as the experimental group with concentrations ranging from 2.250 g/L to 0.281 g/L prepared with trypticase peptone yeast (TPY) culture medium, and TPY culture medium was used as the control group. Ss at the concentration of 10(8) CFU/ml was added to each group, and cultured for 6, 18, 24, 48 hours. Neson-Somogyi method was used to measure the content of reducing sugar, and glucosyltransferase (GTF) activity. The activity of lactate dehydrogenase (LDH) was measured by lactic acid and pyruvic acid continuous monitoring method. The content of water insoluble glucan (WIG) was measured by anthrone method, and the pH value of the culture solution was detected. The value of pH before the experiment and the time difference was alculated as ΔpH.

RESULTSAt the same time point, the activity of GTF and LDH and the concentration of WIG and the value ΔpH decreased gradually with the increase of concentration of LPE. There were significant differences between each experimental group and control group (P < 0.01). The control group had the maximum value, GTF: (6.71 ± 0.61) mIU, LDH: (135.8 ± 1.7) U/L, WIG: (47.15 ± 5.12) mg/L, ΔpH: (2.67 ± 0.01). The highest drug concentration group had the minimum value: GTF: (0.39 ± 0.07) mIU, LDH: (95.0 ± 5.4) U/L, WIG: (2.44 ± 0.38) mg/L, ΔpH: (0.61 ± 0.01).

CONCLUSIONSThe LPE below the MIC could still inhibit the GTF, LDH activity and lead to the decrease of WIG and the acid production.

Dose-Response Relationship, Drug ; Glucans ; biosynthesis ; Glucosyltransferases ; antagonists & inhibitors ; metabolism ; Lactate Dehydrogenases ; antagonists & inhibitors ; metabolism ; Microbial Sensitivity Tests ; Oils, Volatile ; pharmacology ; Plant Oils ; pharmacology ; Streptococcus sobrinus ; drug effects ; metabolism

OBJECTIVETo explore attenuation and mechanism of endoplasmic reticulum stress (ERS)-mediated hepatocyte apoptosis in rats with alcohol-induced liver injury by Qinggan Huoxue Recipe (QGHXR) and its disassembled formulas (Qinggan Recipe and Huoxue Recipe respectively).

METHODSA rat model of chronic alcoholic liver injury was successfully established using a compound reagent of alcohol, corn oil, and pyrazol. The modeled rats were randomly divided into the model group, the QGHXR group, the Qinggan Recipe (QGR) group, and the Huoxue Recipe group (HXR). The CCl4 control group and the normal control group were also set up. There were ten rats in each group. All rats of modeled groups were gastrogavaged with alcohol compound reagent every morning. Rats in the QGHXR group (at the daily dose of 9. 5 g/kg, QGR group (at the daily dose of 3.0 g/kg), and HXR group (at the daily dose of 6.5 g/kg) were administered with corresponding medicines by gastrogavage every afternoon. Equal volume of normal saline was given to rats of the model group by gastrogavage. CCl4 was intraperitoneally injected at the dose of 0.3 mL/kg to rats in the CCl4 control group, once per week. Normal saline was given to rats in the normal control group by gastrogavage. The treatment was lasted for two weeks. Pathological changes of the liver were observed by histopathology. Serum total homocysteine (tHCY) level was detected by ELISA. The hepatocyte apoptosis rate was detected using flow cytometry. The gene and protein expressions of eukaryotic translation initiation factor 2 alpha (elF-2alpha), phosphorylation elF-2alpha (pelF-2alpha), glucose-regulated protein 78 (GRP78), and Caspase-3 in the liver were examined using Real-time PCR and Westen blot respectively.

RESULTSCompared with the normal control group, typical pathological changes of chronic alcoholic liver injury such as steatosis, inflammation, and even fibrosis occurred in model rats. The hepatocyte apoptosis obviously increased, with the apoptosis rate reaching the five-fold of that in normal rats. Besides, early apoptosis dominated. The serum tHCY level significantly increased. The expressions of p-elF-2alpha, GRP78, and Caspase-3 protein obviously increased (P < 0.01). Expressions of GRP78 and Caspase-3 mRNA significantly increased (P < 0.05, P < 0.01). Compared with the model group, the degrees of the liver injury and the hepatocyte apoptosis in the QGHXR group, the QGR group, and the HXR group were significantly alleviated. The serum tHCY level was significantly lowered. The protein expressions of p-elF-2a, GRP78, and Caspase-3 obviously decreased (P < 0.01). mRNA expressions of GRP78 and Caspase-3 obviously decreased in the QGHXR group (P < 0.05, P < 0.01). Only GRP78 mRNA expression obviously decreased in the QGR group (P < 0.05).

CONCLUSIONQGHXR and its disassembled formulas could attenuate ERS-mediated hepatocyte apoptosis in alcohol-induced liver injury rats by lowering the serum tHCY level and expressions of ERS apoptosis correlated factors.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Endoplasmic Reticulum Stress ; drug effects ; Eukaryotic Initiation Factor-2 ; metabolism ; Heat-Shock Proteins ; metabolism ; Hepatocytes ; drug effects ; pathology ; Homocysteine ; blood ; Liver Diseases, Alcoholic ; blood ; drug therapy ; pathology ; Rats ; Rats, Sprague-Dawley

Objective To investigate the current situation and variation in the tendency of Kaschin-Beck disease (KBD) in Shanxi province, to assess the effectiveness of prevention measures, and to provide evidence for making scientific and effective tactics in prevention and control of the disease. Methods In 2008, according to "The National Technical Scheme for Kaschin-Beck Disease Control in 2007", children aged 7 - 12 years underwent clinical and X-ray examination in two historical severe KBD areas of Shanxi province, the observed position on Xray film was finger bone and carpal bone. Adults aged 16 years and above were clinically examined in 50 villages of 13 counties of KBD areas, Shanxi province. The diagnosis was based on "Diagnostic Standard of Kashin-Beck Disease"(GB 16003-1995), the adult patients were degreed according to the state of illness and divided into different groups according to their ages. Results A total of 181 children were examined, none of them was diagnosed as clinical KBD patient. The positive cases of X-ray were 2, X-ray detectable rate of metaphysis was 1.1% for children.In the 50 villages 13 871 of adults were examined and 801 KBD patients were found and the detectable rate was 5.77%. The detectable rate increased with age. The adult patients distributed mainly in the subpopulation aged 51years and above and these patients accounted for 70.66% (566/801) of total people examined. Degrees Ⅱ and Ⅲ patients at these ages accounted for 71.78%(206/287) of total degrees Ⅱ and Ⅲ patients. Conclusions The state of child KBD in Shanxi is under the national control standards. The detectable rate of adult KBD is higher. Relevant departments should pay close attention to the treatment of adult patients.

Objective To explore the relationship between the expression of decoy receptor 3 (DcR3) and high-risk human papilloma virus(HR-HPV)infection in cervical carcinoma. Methods Immunohistochemistry and hybird capture Ⅱ assay were used to detect the expression of DcR3 and HR-HPV in 35 cases of normal cervical tissues(NCE), 39 cases of cervical intraepithelial neoplasm(CIN)and 44 cases of cervical squamous epithelial carcinoma(CSES). Specific HR-HPV16 E7 siRNA and nonspecific HR-HPV16 E7 siRNA were synthesized and transfected to SiHa cells by Lipofectamine. The expression of DcR3 at mRNA and protein levels was examined by real-time polymerase chain reaction and western blot. The growth inhibition was examined by MTT assay. Results In NCE, CIN and CSES, the positive expression rates of DcR3 were 8.6%(3/35), 48.7%(19/39)and 77.3%(34/44), respectively, and the expression intensity was increasing(2=36.942, P<0.001). In NCE, CIN and CSES, the infection rates of HR-HPV were 5.7%(2/35), 56.4%(22/39)and 93.2%(41/44), respectively(2=60.322, P<0.001) . The protein expression of DcR3 was positively correlated with the infection of HR-HPV in CSES(r=0.893, P=0.004). Conclusions DcR3 was highly expressed in cervical cancer. Its expression was positively correlated with HR-HPV infection, which may contribute to the occurrence and development of cervical cancer. HR-HPV silencing inhibited cellular growth and proliferation by down-regulating the expression of DcR3.

Objective To construct the Neisseria surface protein A (NspA) DNA vaccine of Neisseria gonorrhoeae and evaluate the humoral and cellular immune responses induced by this vaccine in mice model.Methods The recombinant expression vector pcDNA3.1 (+)/NspA was constructed by inserting NspA gene into the eukaryotic expression vector pcDNA3.1 (+) and confirmed by poly merase chain reaction (PCR),restriction enzymes HindⅢ,XbaⅠand DNA sequencing.NspA mR- NA in transfected RAW264.7 cells and NspA protein expression in transfected COS-7 cells were de- tected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical stai- ning,respectively.Forty-five male BALB/c mice were immunized with pcDNA3.1 (+)/NspA recom binant plasmid.The level of serum anti-Neisseria gonorrhoeae antibody of the immunized mice was detected by tube agglutination test,and the level of interieron (IFN)-?was assayed by enzyme-linked immunosorbent assay (ELISA).The proliferation of splenocytes was determined by methyl thiazolyl tetrazolium (MTT) colormetry.The NspA gene in BALB/c mice was identified by PCR with the total DNA extracted from quadriceps femoris in immunized sites.Results Restriction enzymes digestion a- nalysis and DNA sequencing results revealed that the pcDNA3.1 (+)/NspA had been constructed successfully.NspA gene had been transcripted and expressed in mammalian cells.The peak titer of specific antibody was 1:640 in pcDNA3.1(+)/NspA immunized group and there was no specific an- tibody detected in both pcDNA3.1 (+) immunized group and PBS group.The IFN-?level in pcD NA3.1 (+) immunized group was (23.79?11.85)pg/mL and that in pcDNA3.1 (+)/NspA immu- nized group was(169.71?30.52)pg/mL (P

BACKGROUNDA new technique developed in 2002, real time endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA), has been one of the most important tools in lymph nodes (LNs) staging before lung cancer surgery. EBUS-TBNA was introduced into China in 2008.

METHODSBetween June 2009 and October 2009, 30 patients with mediastinal/hilar lymphadenopathy and thoracic masses previously detected with CT scan underwent EBUS-TBNA without rapid onsite cytological examination.

RESULTSFrom 30 patients, 33 samples were obtained from LNs and seven samples from intrapulmonary lesions. Twenty out of the 23 lung cancer diagnoses were clarified through the procedure, with sensitivity, specificity, positive predictive value, negative predictive value and accuracy being 87%, 100%, 100%, 70% and 90%, respectively. All three false negative cases were found in the first five procedures. Additionally, among the 33 LNs examined, three specimens that had no lymphocytes were also found within the first five procedures. There were no major complications, and the procedures were uneventful.

CONCLUSIONSEBUS-TBNA seems a safe and effective technique in making diagnosis for mediastinal/hilar LNs and intrapulmonary masses. For pulmonologists experienced in bronchoscopy, the sensitivity of the procedure for diagnosing lung cancer should be no less than 90% after the initial five procedures.

Adult ; Aged ; Aged, 80 and over ; Biopsy, Fine-Needle ; methods ; Bronchoscopy ; methods ; Endosonography ; methods ; Female ; Humans ; Lung Neoplasms ; diagnosis ; pathology ; Male ; Middle Aged

OBJECTIVETo investigate the role of PERK/eIF2alpha signaling pathway in hepatocyte apoptosis of alcoholic liver injury rats.

METHODSRat models with ethanol-induced liver injury were successfully developed by gastric gavage with ethanol-corn oil mixtures for 12 weeks. At different time points (4, 6, 10, 12 week), liver pathology was dynamically observed. The hepatocyte apoptosis was quantitatively analyzed by Annexin V-FITC/PI double-labeled flow cytometry, the serum total homocysteine (tHCY) level was detected by ELISA and the expressions of eIF2a, p-eIF2a, GRP78/Bip, GRP94, caspase-3 and caspase-12 in liver were examined using Real-time PCR and Western blot.

RESULTSTypical acute liver injury and chronic liver injury were observed at week 4 and week 12 respectively. The hepatocyte apoptosis rates in 6-week model rats significantly increased compared with normal rats (P value less than 0.05), and the degree of hepatocyte apoptosis continued to increase with the modeling time, and the percentages of early and total apoptosis reached 26% and 29% at week 12. From week 6 to week 12, the serum tHCY levels in model rats were obviously higher than in normal rats (P value less than 0.01). Since week 4, eIF2a protein phosphorylation in model rat livers remarkably elevated compared with that in normal rat livers (P value less than 0.01), and at week 12 the peIF2a protein expression in model rat livers increased by 2.81-fold. Since week 4 the expressions of GRP78/Bip, GRP94, caspase-12 and caspase-3 mRNA and protein in model rat livers showed a significant increase as compared to normal rat livers, and at week 12, these gene and protein levels increased 4.70, 12.95, 3.83, 4.05 fold and 3.93, 6.93, 9.88, 3.31 fold, respectively (P value less than 0.01).

CONCLUSIONActivation of PERK/eIF2a signaling pathway contributes to the occurrence and development of hepatocyte apoptosis in alcoholic liver injury rats and it might be as a potential target for therapeutic applications in alcoholic liver diseases.

Animals ; Apoptosis ; Eukaryotic Initiation Factor-2 ; metabolism ; Hepatocytes ; cytology ; pathology ; Liver Diseases, Alcoholic ; metabolism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; eIF-2 Kinase ; metabolism

OBJECTIVETo explore the diagnosis and surgical treatment of abdominal cocoon.

METHODSThe clinical data of 16 patients with abdominal cocoon admitted to our hospital between Jun. 1993 and Oct. 2006 were analyzed retrospectively.

RESULTSPreoperatively, Barium meal X-rays revealed coils of intestine in 8 cases, incomplete intestinal obstruction in 6 cases, and prolonged intestinal transit time in 2 cases. CT scan showed dilated intestine and intestinal loops seemed to be encapsulated in a thickened capsule. After opening the peritoneum, entire or partial intestine encapsulated in thickened membrane encasing were found, including partial intestine encapsulated in 1 cases(Type I), entire intestine encapsulated in 2 cases (Type II), and entire intestine and other organs encapsulated in 12 cases(Type III). All the cases underwent adhesiolysis. Intestinal splint was done in 2 patients, gastrostomy in one patient with chronic pyloric obstruction, radical resection of rectal cancer in one patients and ileocolic resection in one patients with Crohn's disease. All patients were healed by surgical operation and confirmed the diagnosis histopathologically.

CONCLUSIONSAbdominal cocoon is rare. It is difficult to make a right diagnosis preoperatively. Barium meal X-rays and CT scan are useful methods for its diagnosis. For the treatment, attention should be paid on complete resection of fibrous membrane, adhesiolysis and prevent intestinal obstruction.

Adolescent ; Adult ; Aged ; Female ; Gastrointestinal Diseases ; Humans ; Intestinal Obstruction ; prevention & control ; Male ; Middle Aged ; Peritoneal Diseases ; diagnosis ; surgery ; Retrospective Studies ; Tomography, X-Ray Computed ; X-Rays ; Young Adult