OBJECTIVEThis study investigates the immune status of T helper (Th) 17 cells in mice with periodontitis.

METHODSSeven-week-old C57BL/6 female mice were randomly divided into the control and periodontitis groups. Each group comprisedfour mice. Experimental periodontitis was induced in mice by oral infection with Porphyromonas gingivalis in the periodontitis group. Phosphate-buffered saline solution was used in the control group. All mice were sacrificed 4 weeks after the last P. gingivalis infection. CD4⁺retinoid-related orphan receptor (ROR) γτ⁺(Th17) cells were analyzed by flow cytometry. The protein expression of Th17 cell-related cytokine interleukin (IL)-17A was detected by enzyme-linked immunosorbentassay (ELISA).

RESULTSFlow cytometry showed that the percentage of CD4⁺RORγτ⁺(Thl7) cells in all CD4⁺ cells and the cell number in gingival tissues, cervical lymph nodes (CLNs), and the peripheral blood obviously increased (P < 0.01) in the periodontitis group. ELISA showed that compared with the control group, the periodontitis group exhibited an obvious increase in the protein expression of IL-17A (P < 0.05).

CONCLUSIONTh17 cell-mediated cell response is enhanced, and the gingival tissues, CLNs, and the peripheral blood are probably the main locations of Thl7 cell-mediated cell response during the development of periodontitis.

Alveolar Bone Loss ; Animals ; Cytokines ; Female ; Flow Cytometry ; Gingiva ; Interleukin-17 ; metabolism ; Mice ; Mice, Inbred C57BL ; Periodontitis ; metabolism ; Porphyromonas gingivalis ; Random Allocation ; T-Lymphocytes, Helper-Inducer

Objective:To investigate regulatory T(Treg)cell-mediated immune response in mice with periodontitis.Methods:8 sev-en-week old C57BL/6 female mice were randomly divided into 2 groups(n =4).Periodontitis was induced in mice by oral infection with Porphyromonas gingivalis(P.gingivalis).Carboxymethyl cellulose(CMC)was used for the sham control mice.All mice were sacrificed 4 weeks after the last P.gingivalis infection.CD4 +forkhead box P3 (Foxp3)+ cells were analyzed by flow cytometry.The protein ex-pression of TGF-1 and IL-1 0 was detected by ELISA.Results:Periodontitis mice showed an obvious decrease in the percentage of CD4 +Foxp3 + cells in CD4 + T cells in gingival tissues,cervical lymph nodes(CLNs)and peripheral blood(P <0.01 ).Moreover,peri-odontitis mice had an obvious increase in the expression of TGF-1 and IL-1 0 in gingival tissues,CLNs and serum(P <0.05).Conclu-sion:Treg cell-mediated immune response is weakened in the mice with periodontitis.Gingival tissues,CLNs and the peripheral blood are probably the main source of Treg cells during periodontitis.

In order to obtain liposomes with properties of heptocyte-specificity and pH-sensitivity,four galactosylated derivatives were synthesized. A series of liposomes were prepared by mixing the galactosylated derivatives with DC-chol/DOPE respectively. The liposome 18-gal was proven to have favorable gene transfer efficiency to human hepatoma HepG2 cells, which was significantly inhibited in the presence of galactose solution, indicating that the liposomal transfection activity was mediated by asialoglycoprotein receptors. The liposome showed prominent pH-sensitivity and low cytotoxicity. Its optimum gene transfer conditions were also determined. The results showed that the liposome may be developed as a potential hepatocyte targeting pH sensitive delivery system for nucleic acid drugs.

Objective:To understand general hospital medical staff status quo and characteristics of blood -borne occupational exposure , analysis its ethics factor , to make the prevention countermeasures of blood -borne oc-cupational exposure to provide scientific basis .Methods:From January 2013 to December floor all the medical staff of blood -borne occupational exposure cases were retrospectively analyzed .Results:A total of 101 medical staff blood-borne occupational exposure , which is given priority to with the nurse , accounted for 60 .40%;More con-centrated in under 30 employees, accounted for 70.29%;Occupational exposure personnel distribution in the ma-jority with surgical department (42.57%), followed by the physician (36.63%);Wards (60.40%), the operat-ing room (15.84%) and therapy (11.88%) are the sites of occupational exposure often happen;Sharp injury in the composition of the ratio of 87.12%, in the first place, in which a proportion (23.76%), pull out the needle (18.81%) and blood (15.84%), surgical suture (14.85%) and transfusion injection (13.86%) as the sharp injury of frequent occurrence of occupational exposure;Exposure is given priority to with hepatitis b ( 61 .38%) . Conclusion:Medical staff blood-borne occupational exposure risk is high , the hospital infection control personnel must pay attention to occupational exposure of the whole education , strengthen the administration of the occupation-al exposure of ethics , reduce the risk of occupational exposure and injury .

Objective To establish a method for detecting K-rag gene point mutation in pancreatic adenocarcinoma based on the pyrosequencing and to compare its performance with that of Sanger sequencing.Methods Genomic DNA was extracted from formalin-fixed,paraffin-embedded pancreatic tissues including 49 pancreatic adenocarcinoma,10 normal pancreas,11 chronic pancreatitis,18 benign pancreatic tumor,7 insulin carcinoma,9 ampullary carcinoma,7 bile duct carcinoma and 7 duodenum papillary adenocarcinoma tissues.K-rag gene point mutations at codon 12 were analyzed by pyrosequencing and Sanger sequencing,respectively.Results No mutant K-ras gene Wag detected in normal pancreas,chronic pancreatitis,benign pancreatic tumor,insulin carcinoma,ampullary carcinoma,bile duct carcinoma and duodenum papillary adenocarcinoma tissues by either pyrosequencing or Sanger sequencing.K-rag gene point mutation was detection rate in pancreatic adenocarcinoma tissues was 71.4%(35/49)by pyrosequencing and 61.2%(30/49)by Sanger sequencing,respectively.Conclusions Pyrosequencing is more sensitive than Sanger sequencing and is also accurate,rapid and of high throushput in detecting mutant K-ras gene.It may serve as a practical method for fast batch analysis of clinical samples.

ObjectiveTo investigate the effect of heart-regulating and mind-calming acupuncture on the expression of gap junction protein connexin43 (Cx43) in myocardial cells in a rabbit model of ventricular arrhythmia.MethodA rabbit model of ventricular arrhythmia was made by injecting barium chloride into the marginal ear vein. Forty New Zealand rabbits were randomized into control, model, acupuncture and medication groups. The acupuncture group received heart-regulating and mind-calming acupuncture[at Lingtai(GV10), Shendao(GV11), bilateral Neiguan(PC6)and Baihui(GV20)]and the medication group, oral gavage of propafenone. Rabbit left ventricle was taken after the completion of the experiment. Cx43 expressions were compared and analyzed usingan immunohistochemical staining technique and a computer image processing system.ResultCx43 expression decreased significantly in the model group compared with the control group (P<0.01) and increased in the acupuncture and medication groups compared with the model group (P<0.05). There was no significant difference in Cx43 expression between the acupuncture and medication groups (P>0.05).Conclusion Adjusting Cx43 expression in myocardial cells may be one of the target points for this acupuncture treatment of arrhythmia.

Objective To explore the change of the pre-swallow peak pressure of upper esophageal sphincter (UES) in patients with post-stroke cricopharyngeal achalasia,and investigate the effect of pre-swallowing peak UES pressure on swallowing function by quantitative analysis.Methods Fifty-seven stroke patients with cricopharyngeal achalasia were recruited and divided into balloon dilation group,combined training group and routine swallowing training group with 19 patients in eachp.All the three groups accepted routine swallowing training.In addtion,the routine swallowing training group and balloon dilation group accepted larynx elevation training and balloon dilation training,respectively,while the combined training group accepted larynx elevation training and balloon dilation training simultaneously.The pre-swallow peak UES pressure was measured by using PC polygraph high rate gastrointestinal dynamical detection system (PC Polygraf HR,CTD-synectics,Sweden) before and after 8 weeks of treatment.The swallowing function was assessed using swallowing function classification and water swallowing test.Results Before treatment,there was no significant difference among the 3 groups in terms of the pre-swallow peak UES pressure,swallowing function classification,water swallowing test and VFSS (P ＞ 0.05).After treatment,pre-swallow peak UES pressure,swallowing function classification,water swallowing test and VFSS of the balloon dilation group and combined training group improved significantly compared with those before treatment (P ＜ 0.05),and the improvement in the combined training group was to a significantly better extent than in the balloon dilation group(P ＜O.05).Conclusion Balloon dilation and larynx elevation training plus routine swallowing training can increase pre-swallow peak UES pressure,decrease the UES resting pressure of stroke patients with cricopharyngeal achalasia,which is of great importance for their recovery.

Objective To observe the influence of transplanting neural stem cells (NSCs) on angiogenesis in rats with spinal cord injury (SCI).Methods The Allen's method was used to create SCI models in sixty adult Sprague-Dawley (SD) rats.They were then randomly classified into a control group which received injections of phosphate buffered solution (PBS) and an NSC group which received injections of NSCs via the tail vein,with 30 rats in each group.Another group of 30 similar rats without SCI received injections of NSCs via the tail vein as the normal group.Each rat was evaluated before transplantation and at days 7 and 14 post-transplantation using the Basso,Beattie and Bresnahan (BBB) scale for testing hindlimb function.After sacrifice,the distribution of yon Willebrand factor (vWF) in both groups was determined by immunofluorescence,and Western blotting was used to detect vascular endothelial growth factor (VEGF) protein.Results The average BBB score of the normal group was 21 at every time point.Before transplantation,the BBB scoresof the control and NSC groups were both 0,however they increased over time.At day 7 post-transplantation,the BBB scores showed no significant difference between the control group and the NSC group.At day 14 post-transplantation,the average BBB score of the NSC group was significantly higher than that in the control group.At days 7 and 14,the counts of vWF-positive cells in the normal group were significantly higher than in the control and NSC groups.VEGF protein expression in the normal group was significantly lower than in the NSC and control groups.Conclusions NSC transplantation may promote angiogenesis after spinal cord injury and improve motor function by inducing the expression of VEGF.

Objective: To establish a method for indentification of the podophyllum emodi species, Radix Clematis and Radix Gentiana species. Methods: HPLC/UV fingerprint analysis method of toxic ingredient podophyllotoxin and its derivatives were developed, and the method had been evaluated. Results: The methodological evaluation showed that this method had a good repeatability and reproducibility, and different samples had different HPLC fingerprints. Conclusion: This method can be used to differentiate podophyllum emodi from two commonly used medicinal herbs of a different genus but having similar appearance, Radix Clematis and Radix Gentiana.