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 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

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 We investigated the changes of the motorial network in patients suffered from brain tumors adjacent to the central sulcus occurred with reorganization of motor function using function connectivity MRI(fcMRI)technique in order to provide the new evidence for the compensational hypothesis of the reorganization caused by focal lesions.Methods Using 1.5 T MRI unit,14 patients with brain tumors in the vicinity of the central sulcus occurred with reorganization of motor function and 6 normal volunteers were examined with fcMRI technique while the subjects performed no task.By selecting seed voxels(region of interest)in the regions showing the most activation in M1 area on the activated map and cross correlating with every voxel within the brain,the fcMRI maps based on unilateral primary motor(M1)area were calculated.The location,extent and volume of the region showing significant connectivity to the several seed voxel,such as left/right M1 area in the health group and affected/unaffected M1 area in the patient group were evaluated on the fcMRI map.Results In healthy group,the extent and volume of the region showing significant connectivity to the left M1 area[(9514.17±186.92)mm3]were almost similar to those to the right M1 area [(9364.67±382.75)mm3].There showed no significant difference in motor connectivity between the two groups(P>0.05).In the tumor group,the volume of regions showing significant connectivity to the M1 area located in the affected hemisphere [(11193.14±811.29)mm3]was obviously higher than that of regions based on the seed voxel in the unaffected side[(6549.86±400.94)mm3](t=20.383,P＜0.01).The volume was significantly different among the regions showing high connectivity to the M1 of the affected side in patient group.those showing significant connectivity to the left M1 and fight M1 in health group(P＜0.01),the former was the biggest(P＜0.01).The extent of the regions showing connectivity to the affected M1 was consistent with the reorganization area of motor function revealed by fMRI.The volume of regions showing significant connectivity to M1 area of unaffected hemisphere in patient group showed significant difference compared with those showing significant connectivity to the left M1 and right M1 in health group(P＜0.01),the former was smallest(LSD,P＜0.01).especially in the affected hemisphere.It might mean the disrupted functional connectivity between the M1 area of unaffected hemisphere in patient group and motor area located in the collateral side.Conclusion The increased connectivity between the M1 area of the affected hemisphere and the other motor cortex might indicate that the reorganization in the motor pathway and the formation of the potential compensatory network second to the impairment of the normal motor pathway resulted in the functional reorganization of the motor cortex.The fcMRI technique might be a valuable approach to reveal the pathophysiological changes of nerve network caused by brain tumor.

OBJECTIVETo study the clinical significance of expression of hepatic growth factor (HGF) and its receptor c-Met in renal cell cancer (RCC).

METHODSTwenty-six patients with RCC and 10 benign renal tumor patients were examined. The expression of HGF and c-Met mRNA was detected using Northern blot. GAPDH was used as the internal control.

RESULTSTwenty-one out of 26 patients with RCC had positive gene expression of HGF and c-Met with the positive rate of 80.8%. According to the tumor TNM staging, the more advanced the cancer had a stronger expression of HGF and c-Met, the expression of c-Met was higher than that of HGF in the RCC. Whereas in the control group with benign tumor, the expression of c-Met was rather mild.

CONCLUSIONThe result suggests that HGF and its receptor c-Met may play an important role in the development and progression of renal cancer, which may also imply a potential clinical significance in diagnosing and assessing prognosis of RCC.

Adult ; Aged ; Carcinoma, Renal Cell ; metabolism ; pathology ; Female ; Hepatocyte Growth Factor ; biosynthesis ; genetics ; Humans ; Kidney Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Proto-Oncogene Proteins c-met ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis

OBJECTIVETo explore a method for ectopic prefabrication of mandible with vascular pedicle.

METHODSCancellous bone blocks harvested from the dog ribs were packaged with mandible-shaped titanium mesh scaffold and implanted into latissimus dorsi of dog with thoracodorsal artery and vein through the scaffold. After 12 weeks, bone formation and vascularization were evaluated by gross inspection, histological examination and immunohistochemistry.

RESULTSVascularized mandible with thoracodorsal artery and vein were formed and histological staining and immunohistochemisty confirmed new bone formation and vascularization.

CONCLUSIONSThere is feasibility for ectopic prefabrication of vascularized mandible graft using cancellous ribs, which provides a new method for mandibular defect reconstruction. Experimental study on ectopic prefabrication of vascularized mandible graft with autogenous ribs.

Animals ; Bone Transplantation ; methods ; Dogs ; Female ; Male ; Mandible ; surgery ; Osteogenesis ; Reconstructive Surgical Procedures ; methods ; Ribs ; surgery ; Tissue Engineering ; methods ; Transplantation, Autologous