砄bjective:To analyze the DNA polymorphism of Prevotella intermedia(P.i ) and their distribution at diseased and healthy sites in adult patients with periodontitis.Methods:101 P.i strains were obtained from subgingival plaque of the adults with periodontitis.Aabitrarily primed polymerase chain reaction (APPCR)with OPA 03 and OPA 13 as primers was used to study the genetype of P.i .Results:The OPA 03 primer AGTCAGCCAC grouped 101 P.i strains into 7 different genotypes,while OPA 13 CAGCACCCAC divided 97 P.i strains into 12 genotypes.There was mainly one genotype of P.i in each subject,and mainly one genotype of P.i in individual diseased and healthy site.Also,several different genotypes of P.i were found in a identical subject and at a identical site.The predominant genotypes of P.i in diseased sites were mostly the same to that in healthy sites,no special genotype of P.i which was associated to periodontal status was found.Conclusion:Overgrowth of P.i without special genetype may be a pathogen for the pathogenesis of periodontitis.

Objective To investigate the clinical and laboratory characteristics of hematological malignancies with t(1 1;19)(q23;p13.1).Methods Chromosome specimens of bone marrow cell from a patient with hematological malignancy were collected.After short-term culture,and chromosome karyotype analysis was carried out by R banding technique.Results The chromosome karyotype of the patient diagnosed with AML M4+ was t (11;19)(q23;p13.1).The patient did not obtain complete remission after application of MA regimen chemotherapy.Conclusions t(11;19)(q23;p13.1) translocation is a rare and recurring chromosome abnormality,which is related with a specific type of AML.The prognosis of the AML patients with this chromosome abnormality is poor.

Objective:To evaluate the effects of periodontitis on the expression of apoptosis-related proteins in pancreas of rats with Type 2 Diabetes Mellitus.Methods:Spontaneously type 2 diabetic OLETF rats were randomly divided into 2 groups:diabetes with or without periodontitis(diabetes group and combination group).LETO rats with the same germline and the same age but having normal glucose tolerance were randomly divided into control group and periodontitis group.20 weeks after periodontitis were established,all the rats were sacrificed and the pancreas were pathologically examined by HE staining.The expression of Bax,Bcl-2 and Caspase-3 in the pancreas islet were detected by immunohistochemistry staining and semi-quantitative analysis.Results:The expression of Bax, Bcl-2 and Caspase-3 in the pancreas islet was no significant difference between control and periodontitis groups(P=0.324,P=0.091,P=0.852).Compared with diabetes group,the expression of Bax and Caspase-3 in combination group showed a significant increase(P=0.000,P=0.000),and the expression of Bcl-2 was significantly decreased(P=0.022).Conclusion:Under healthy conditions,periodontitis has no effect on the expression of apoptosis-related proteins in rat pancreas islet.However,in rats with diabe-tes,periodontitis may affect the expression of apoptosis-related proteins in pancreas islet.

Objective　To investigate the surgical management of a gun shot wound of blood vessels and immersied, and evaluate its primary effect. Methods　100 rabbits were divided randomly into simple wounded group(SWG,n=50) and seawater immersion group(SIG,n=50).F emoral arteries were impacted by 0.38 gram steel spheres with velocity of 600 ～800 meters per second fired by 7.62 mm rifle. Animals in SIG were immersed in artificial seawater (pH 8.2～8.4, salinity 25.4,temperature 21℃) for 60 min, o f which those in SWG were spared. Grossly injuried artery was excised and restor ation of blood flow was reconstructed by end-to-end anastomosis or reversed au togenous venous grafting or cryopreserved arterial allografting. At 24 h,7,1 4,21 days after operation, blood flow was examined by Doppler ultrasonic detecti on and part of anastomotic sites and graft were collected for pathological obser vation. Results　In completely transected injury, the patency in SIG was 80.00%,while that in SWG was 86.67% in the first 3 weeks. In arterial c ontused injury ,patency in SIG was 86.67%,and that in SWG was 82.35% at the same time. Thrombosis occurred mostly in the first postoperative week. Atypical endo thelial cells were found at the anastomosis sites in the first postoperative week, and the anastomosis sites were lined with endothelium in 3 weeks postopera tively. Conclusion　Early curative effect could be obtained. Whe n grossly injuried artery is excised and followed by a routine surgical procedur e in the treating gunshot wounds immersed in seawater.

Objective To evaluate the value of high-frequency ultrasound and X-ray barium meal examination in diagnosis and typing diagnosis of congenital hypertrophic pyloric stenosis(CHPS).Methods High-frequency ultrasound and X-ray barium meal examination were made in newborns with present symptom of vomiting,and comparison was made between the two examine methods.Retrospective analysis was made in 29 cases confirmed by surgical operation.Control normal group included twenty healthy newborns.Results In 29 CHPS cases,the diagnostic accordance rate of ultrasound was 1 00%.And the diagnostic accordance rate of X-ray barium meal examination was 93.1%(27/29).In all those CHPS cases,the length of pyloric canal were≥18 mm,the diameter of pyloric canal were≥14 mm,the wall thickness of pyloric canal were≥4 mm,the stenosis inside diameter were≤6 mm by ultrasound;in X-ray barium meal examination,the length of pyloric canal were≥20 mm,the inside diameter were≥7 mm,and there was significant difference in comparison with control group(P＜0.01).The CHPS was classified into two groups as mild stenosis and severe stenosis according to the inside diameter and the wall thickness of pyloric canal.The mild type showed the wall thickness was＞4 mm and≤6 mm,the inside diameter was＞3 mm;The severe type showed the wall thickness was＞6 mm,the inside diameter was≤3 mm or obliterate.Conclusions High-frequency ultrasound has significant clinical application value in diagnosis and subtype diagnosis of CHPS,it is an easy,reliable and safe examination method.

Objective To investigate the expressions of tetraspanin CO-029 in intrahepatic cholangiocarcinoma (ICC) and to find out their clinical significance.Methods RT-PCR and western blot were used to detect the expressions of CO-029 in ICC and their matched para-tumorous tissues from 20 patients with ICC,as well as in the HCCC-9810 cell lines.The expressions of CO-029 were further detected via tissue microarray (TMA) in the pathological specimens of 40 patients with ICC.Correlations between the expressions of CO-029 and the clinicopathologic features and prognosis were analyzed.Results A high level of CO-029 was detected in the 20 patients with ICC and the HCCC-9810 cell lines via western blot and RTPCR.Moreover,the expression levels of CO-029 in the ICC tissues were higher than the matched para-tumorous tissues (P ＜ 0.05).TMA detection revealed the positive expression rate of CO-029 to be 65％ (26/40).The expression level of CO-029 was much higher in the early recurrence group (Time to recurrence,TTR ＜ 1 year) than the non-recurrence group (TTR≥ 1 year).On analysis,the correlations were significant between the expressions of CO-029 and tumor encapsulation,hilar lymph node metastasis,TNM stage and prognosis (P ＜0.05).Conclusions CO-029 was highly expressed in ICC.It had close correlations with recurrence,metastasis and prognosis of patients with ICC.

Objective To search for a good solution for aesthetic beard reconstruction and to explore the effect of the single-hair grafting in beard reconstruction. Methods Under the local anesthesia, a scalp strip (length range from 5～7 cm, width range from 1.0～1.5 cm) was harvested from the back of the head and then was divided into a series of single-hair micrografts. After the slits were made, the grafts were implanted into the holes, parallel to the original beard directions. The donor sites were directly closed. With this method, 18 sparse beard patients and 3 patients with upper lip scars were treated. Results The grafted hairs grew nearly in the direction of normal beard, which achieved 90 %～95 % survival in recipient areas. The patients were satisfied with the results, in which 19 cases received only 1 operation and 2 cases received another repeated operation. Conclusion Beard reconstruction by single-hair grafting technique is a good, safe and effective method.

Objective To investigate the role of transmembrane protein C0-029 in epithelial mesenchymal transition (EMT) induced by tumor necrosis factor alpha (TNFα) of intrahepatic cholangiocarcinoma (ICC) cells.Methods RT-PCR and Western blot were conducted to detect the CO-029 expression in ICC cells and tissues.The effect of CO-029 silencing by lenti-virus on EMT induced by TNFα was investigated.Western blot and mass spectrometry after immunoprecipitation were used to confirm whether TNFαR1 can directly or indirectly bind with CO-029 to form complexes in ICC.Results Differential expression of CO-029 was observed in ICC cells and tissues.The expression of CO-029 was significantly reduced by lenti-viral interference in ICC cells,resulting in the failure of TNFα to induce EMT.TNFαR1 in ICC could directly or indirectly form complexes with CO-029.Conclusion CO-029 mediates TNFα/TNFαR1 induced EMT in ICC,which might play an important role in the invasion and metastasis.

Objective To investigate the effect of occult HBV infection (OBI) on carcinogenesis of cryptogenic hepatocellular carcinoma.Methods Samples of hepatocellular carcinoma (HCC) and pericarcinomatous tissues obtained after hepatectomy from January 2011 to November 2013 at the Third Affiliated Hospital of Sun Yat-Sen University were collected.They were divided into two groups:the cryptogenic HCC group (the CH group,n =26) and the HBV related HCC group (the HH group,n =40).These samples were compared with the normal liver tissues obtained in 30 patients.HBV DNA was identified by the nested polymerase chain reaction and the immunohistochemical method was taken to examine the hepatitis B virus X protein (HBx) and Yes-associated protein (YAP) expressions.Results OBI was identified in 20 (77.8％) cryptogenic HCC patients and 8 (26.7％) in the control group.There was a significant difference between the two groups (x2 =14.072,P ＜ 0.05).HBV DNA was detected in all the HBV-related HCC patients.The HBx protein expression was mainly located in the cytoplasm of liver cells and liver cancer cells,but YAP was expressed in the nucleus.Both of them showed diffuse brown or tan particles.In the HH group and the CH group,the positive expression rates of HBx protein in the tumorous tissues were 80.0％ and 90.0％,respectively,and 85.0％ and 82.5％ in the nontumorous tissues,but only in 40.0％ in the control group.The positive expression rates of YAP in the tumorous tissues were 65.0％ and 67.5％,respectively,15.0％ and 20.0％ in the nontumorous tissues,respectively,but only in 12.5％ in the control group.The HBx expression in the cancerous tissues and para-cancerous tissues of the HH group and the CH group showed no significant difference (P ＞ 0.05),but the YAP expression in the tumor tissues was significantly higher than that in the nontumorous tissues (P ＜ 0.05).The HBx and YAP expressions in the HH group were comparable to the CH group (P ＞ 0.05).However,their expressions in the cancerous tissue of the HH group and the CH group were significantly higher than in the normal liver tissues (P ＜ 0.05).Conclusion A high prevalence of HBV infection was observed in HBsAg-negative HCC and the high expressions of HBx and YAP might be involved in the process of cryptogenic hepatocellular carcinoma.

Objective:To investigate the correlations between CO-029 expression and cholangiocarcinoma invasion and metastasis, and the further explore the potential mechanism involved.Methods:The constructed lentiviral vector of vshRNA-CO-029 (LV/GFP/CO-029) was used to transfect and screen the stable transfected cholangiocarcinoma cell line HCCC-9810-vshRNA-CO-029 as the silence group, HCCC-9810 cells transfected with the mock plasmid were used as the mock group, and the untransfected cells were used as the control group. Cell scratch assay, Transwell assay and in vivo implantation assay were used to detect the migration, invasion and metastasis of the three groups of cells. Immunoprecipitation and tumor necrosis factor (TNF)-α stimulation assay were used to detect the effect of CO-029 on the expression of EMT-related genes.Results:The scratch healing rate of the silence group was (27.11±4.58)%, which was lower than that in mock group (92.84±6.24)%, the number of cells passing through Matrigel in silence group was (57.15±6.10), which was significantly lower than that in mock group (108.20±9.21) and control group (112.00±10.45), the differences were statistically significant ( all P<0.01). The volume of liver tumors in the silence group of orthotopic xenograft mouse model was (2.17±0.54) cm 3, while the volume of liver tumors in the transplanted simulation group was (0.74±0.15) cm 3, the differences were statistically significant ( P<0.05). The incidence of lung metastasis and the number of lung metastases in the simulated group was 100%(6/6) and (214.17±35.64), respectively, while that in the silence group was 16.7% (1/6) and (41.56±14.15), respectively, the differences were statistically significant (all P<0.05). Co-immunoprecipitation showed that CO-029 can form a complex with TNF-αR1. TNF-α induced the down-regulation of E-cadherin and up-regulation of vimentin and N-cadherin in the mock group, but no significant changes were observed in the silence group. Conclusion:CO-029 expression is positively correlated with tumor invasion and metastasis of cholangiocarcinoma, and could couple with TNF-α to induce EMT, which is a novel well-established potential prognostic and therapeutic target for cholangiocarcinoma metastasis and prognosis intervention.