Objective:To find out the existing pattern of HPV16 in tongue cancer cell and to analyze itsrole in carcinogenesis of tongue cancer. Methods :Southern blot hybridization was used to detect the HPV16sequence and its existing condition in 20 cases of fresh human tongue cancers. Results:HPV16 DNA se-quence in 5 cases of 20 tongue cancers was detected. And HPV16 DNA existed in tongue cancer cell in non-integration pattern. Conclusion:HPV16 was involved in carcinogenesis of tongue cancer through interactionof HPV gene products rather than its integration with genome of target cell.

Objective: To clone cDNA of enamel matrix serine proteinase (EMSP1) encoding mature protein from mouse dental germs. Methods: Total RNA was isolated from developing incisors and molars of 7 days mouse pups and reverse-transcribed into cDNA. Two pairs of specific primers was designed to obtain the desired gene by Touchdown PCR and Nested PCR. The segment was inserted into Vector pMD-18T, and recombined vectors was transformed into E.coli JM109.The positive clone was chose and analysed by restriction endonuclease mapping and DNA sequencing. Results:700 bp of cDNA of mouse EMSP1 was sueccessfully cloned from mouse tooth germs tissue. The sequence was consistent with that displayed in PubMed. Conclusion:The mouse EMSP1 cDNA encoding mature protein is obtained for further study.

Objective:To study the association of TGF?2 polymorphisms and maternal smoking with the occurrence of nonsyndromic cleft lip and palate(NSCLP). Methods:TGF?2 genes were amplified from peripheral leukocytes by means of PCR in 272 cases of nonsyndromic cleft lip with or without palate(CL/P), 251 of cleft palate only(CPO) and 312 of unrelated controls in Jilin Province, PCR products were analyzed by single-stranded conformation polymorphism(SSCP) and DNA sequencing. Maternal smoking was investigated. The association of TGF?2 polymorphisms, maternal smoking with the occurrence of CL/P and CPO was analyzed by SAS statistic system. Results:The 322 bp PCR product of TGF?2 was amplified from CL/P, CPO and control samples; SSCP analysis showed three alleles of TGF?2;sequencing results showed that allele1, allele2 and allele3 contained seven, eight and nine ACA repeats respectively. The statistic analysis showed that TGF?2 polymorphisms or maternal smoking was associated with the occurrence of CL/P and CPO respectively(P0.05).Conclusion:TGF?2 polymorphisms and maternal smoking during pregnancy are associated with the occurrence of CL/P and CPO. TGF?2 polymorphisms have no interaction with maternal smoking.

Objective To investigate the effects of schistosomiasis on malignant bio-behaviors including histological types,differentiation grade,inasie depth,lymphnode metastasis and prognosis of colorectal carcinoma among elderly people in the district of petrochemical industry,Shanghai.Methods A total of 184 cases (aged 60 years and over) with specimens of colorectal cancer were collected from January 2004 to December 2010.These specimens were divided into colorectal carcinoma with schistosomiasis (CRCS) group (102 cases) and simple colorectal carcinoma (CRC) as control group (82 cases).The prognostic data of 40 elderly cases with colorectal carcinoma from January 2004 to December 2006 were simultaneously selected and analysed Results Totally 55.4％ (102/184) cases exhibited coexistent schistosomiasis with colorectal carcinoma.lesion site(x2=16.53),growth pattern(x2 =10.81 ),differentiation degree(x2 =10.46),infiltration degree(x2 =6.71),lymph node metastasis(x2 =3.90),and TNM staging(x2=5.73) in CRCS group were closely related to schistosomiasis compared with CRC group (P＜0.05).Survival curve analysis showed that there was no significant effect of schistosomiasis on the survival time of elderly patients with colorectal carcinoma (x2 =0.14,P＞0.05).Conclusions The coexistent schistosomiasis may be associated with the malignant bio-behaviors of colorectal carcinoma among the elderly,but have no significant influence on the survival time after operation.

Objective To investigate the effects of obesity on blood pressure variation and plasma levels of nitric oxide (NO) and endothelin (ET) in elderly hypertensive patients.Methods A total of 175 elderly patients with hypertension were screened for this study.Based on body mass index,they were categorized into three groups with normal weight (n =69),overweight (n=56) and obesity (n=50).24 hour dynamic blood pressure,NO and ET levels were monitored.Results No significant differences in 24 h systolic blood pressure,daytime systolic blood pressure,nighttime systolic blood pressure,24 hours diastolic blood pressure (24 h-DBP) and night time diastolic blood pressure were found among the groups (all P＞0.05).Morning systolic and diastolic blood pressure were higher in obese group than in normal weight and overweight groups(both P＜0.05).The daytime diastolic blood pressure was higher in obese group than in overweight group.24 h systolic blood pressure variation,daytime systolic blood pressure variation,and blood pressure pattern were higher in obese group than in overweight and normal weight group [(12.6 ± 2.7)％ vs.(10.4 ±2.2)％ and (9.4±1.9)％,(12.2±2.9)％ vs.(10.2±2.2)％ and (9.2±2.1)％,(5.2±10.5)％vs.(1.4± 6.9) ％ and (1.8 ± 8.2) ％ group,all P＜ 0.05].The nighttime systolic blood pressure variation,24 h diastolic blood pressure variation,daytime diastolic blood pressure variation were increased in obese group as compared with normal weight group [(9.8 ± 3.7)％ vs.(8.2 ± 3.1)％,(15.3±3.3)％ vs.(13.2±4.2)％,(14.7±3.7)％ vs.(12.9±3.8)％,all P＜0.05].No differences were found in nighttime diastolic blood pressure variation among the groups.Plasma NO level was lower in obese group than in overweight and normal weight group [(29.8± 14.2)μmol/L vs.(47.9± 18.6) μmol/L and (94.6 ± 42.9) μmol/L,P＜0.01].Plasma ET level was significantly higher in obese group than in overweight group and normal weight group [(46.5± 9.8)ng/L vs.(37.3±4.8) ng/L and (31.1± 5.5) ng/L,P＜0.01].24 h systolic blood pressure variation was significantly correlated with plasma NO level (r =0.340,P =0.004) in normal weight group.Conclusions Obesity can effectively increase blood pressure variation and ET level,and reduce plasma NO level in elderly hypertensive patients.Obesity is one of the most important influencing factors for blood pressure variation,plasma NO and ET levels.

Objective:To study the application of Gomori special staining in the late bell stage of dental germ,tooth and periodontium.Methods:Paraffin-embedded sections of human dental germ in late bell stage,tooth and periodontium of mature rats were stained by improved Gomori and Mallory special staining method.Results were observed by light microscope.Results:Improved Gomori special staining method can easily discriminate enamel matrix and dentin matrix in late bell stage of dental germ,sclerous tissues and soft tissues in mature tooth and periodontium,bone and cartilage in two different colouration.Cells and tissues can be observed clearly.Conclusion:Dental germ,tooth and periodontium can be stained by improved Gomori special staining with satisfactory results.

Objective:To investigate the effect of poly (lactic acid)(PLA)electrospun membranes loaded with cisplatin and chloroquine on the oral squamous cell carcinoma CAL-27 cells,and to explore the method to prevent the recurrence of oral cancer.Methods: The DDP/PLA membranes, CQ/DDP/CQ/PLA membranes and CQ/DDP/PLA membranes were prepared by electrospinning.Then the micro morphology of three kinds of membranes were observed by scanning electron microscope (SEM);the degradation rate of PLA membrane was measuredby UV spectrophotometric.The LC3-Ⅱ expression level in CAL-27 cells was detected by laser scanning confocal microscope.The survival rate of CAL-27 cells was detected by MTT method.Results:The SEM results showed that the nanofibers of DDP/PLA,CQ/DDP/PLA and CQ/DDP/CQ/PLA membranes were continuous and smooth with uniform diameters.The degrated time of membranes was about 21 d.The MTT result showed that compared with control group,at first,the effects of cell killing of DDP/PLA membranes,CQ/DDP/CQ/PLA membranes and CQ/DDP/PLA membranes were not obvious;as the extension of time,the survival rates of CAL-27 cells in DDP/PLA membranes group,CQ/DDP/CQ/PLA membranes group and CQ/DDP/PLA membranes group were decreased (P <0.05).The immunofluorescence results showed that the fluorescence intensity of LC3-Ⅱ in CQ/DDP/CQ/PLA membranes group and CQ/DDP/PLA membranes group were lower than that in DDP/PLA membranes group.Conclusion:CQ/DDP/PLA membranes with sustained-release effect can increase the sensitivity of CAL-27 cells to DDP and enchance the killer effect of DDP on the CAL-27 cells.

Objective: To use autophagy inhibitors combined with radiation to treat the oral squamous cell carcinoma CAL-27 and KB cells,and to explore the influence of autophagy in the oral cancer radiation sensitivity and its mechanisms. Methods:The human oral squamous cell carcinoma CAL-27 and KB cells were divided into control group,CQ group,3-MA group,IR group,CQ+IR group,and 3-MA+IR group. The survival rate was detected by MTT method and the autophagy of CAL-27 cells was observed by immunofluorescence method and laser scanning confocal microscope.The expression levels of LC3 and beclin-1 were detected by Western blotting method. The apoptotic rate was determined by flow cytometry with Annexin Ⅴ/PI doulde staining. Results:Compared with IR group,the survival rates in 3-MA + IR and CQ+ IR groups were signifcantly decreased (P < 0.05 ).The autophagy levels of cells in IR group were significantly higher than those in control group, CQ group, 3-MA group,CQ+IR group,and 3-MA+IR group (P <0.05).The expression levels of LC3 and beclin-1 proteins in IR group were significantly higher than those in control group,CQ+ IR group,and 3-MA+ IR group (P < 0.05). The apoptotic rates in IR,3-MA+ IR,and CQ+ IR groups were markedly higher than those in control group. Compared with IR group,the apoptotic rates in CQ+IR and 3-MA+IR groups were significantly increased (P <0.05).Conclusion:Radiotherapy can induce the increase of autophagy level of oral squamous cell carcinoma cells. Inhibiors of autophagy can increase the radio-sensitivity of oral squamous cell carcinoma cells by inhibiting the proliferation and inducing the apoptosis.

Objectve To evaluate the serum level of antimicrobial peptide human cationic antimicrobial protein 18 (hCAP18) in colorectal patients and it auxiliary diagnosis and prognosis value.Methods Case-control study was used.The serum level of hCAP18 was measured by enzyme linked immunosorbent assay(ELISA) in 68 cases with colorectal patients of department of gastrointestinal surgery and 40 cases healthy people of department of physical examination from January 2014 to Junc 2015 in Tongji Hosptial of Tongji University.The concentrations of hCAP18 in serum of colorectal patients before and surgery were analyzed.Immunohistochemistry was used to detect hCAP18 expression in colorectal carcinoma.The effect of hCAP18 on colon carcinoma cell proliferation was detected by BrdU-ELISA and soft agar colony formation assay.The sensitivity and specificity of serum hCAP18 for the diagnosis of eolorectal were evaluated using the receiver operating characteristic curves(ROC).Date was analyzed by using the ttest and one-way analysis of variance.Results hCAP18 serum levels in colon cancer of stage Ⅰ,Ⅱ,llⅢ and Ⅳ patients were (0.46 ± 0.18) mg/L,(0.65 ± 0.45) mg/L,(1.26 ± 0.68) mg/L and (2.35 ± 1.06)mg/L.Mean value was(1.16 ±0.88) mg/L,which was significantly higher than in normal people (0.19 ±0.07) mg/L (t =5.290,P ＜ 0.05).hCAP18 levels had significantly decreased in serum of colorectal patients after 30 d surgery compared to preoperative results [from (1.16 ± 0.88) mg/L to (0.26 ± 0.06) mg/L;t =3.971,P ＜ 0.05].Immunohistochemistry results showed hCAP18 was high expression in colon cancer tissue compared with adjacent tissues;BrdU-ELISA assay results showed HCTll6 and SW480 cell proliferation increased significantly after 0.05-1 mg/L of hCAP18 treatment;Soft agar clone formation experiment proved hCAP18 could significant enhance clone formation of HCT116 and SW480 colon cancer cell lines.The size of clonal cluster of HCT116 was increased from (145.40 ± 35.20) μm to (370.80 ± 32.65) μm (t =10.50,P ＜ 0.05) and SW480 was increased from (101.00 ± 27.10) μm to (369.00 ± 27.29) μm (t =15.58,P ＜0.05);The numbers of clonal cluster of HCT116 was increased from 8.50 ± 2.30 to 42.80 ± 6.60 (t =3.945,P ＜ 0.05) and SW480 was increased from 6.20 ± 1.70 to 46.00 ± 7.20 (t =4.775,P ＜ 0.05).ROC analysis of serum hCAP18 yielded an AUC (area under the ROC curve) of 0.93 (95％ CI =0.859-0.999)with 91.17％ sensitivity and 80.00％ specificity,which was higher than the CEA[0.78 (95％ CI =0.699-0.933)].Conclousions Detection of serum hCAP18 shows a good sensitivity and specificity for the auxiliary diagnosis of colon cancer.It is possible to be potential detection index for noninvasive diagnosis and monitoring progression of colon cancer.hCAP18 could promote the proliferation of colon cancer cells,it played an important role in the progression of colon cancer.

This study explored the possibility of using event-related potentials (ERP) for the measurement of picture-recognition memory and examined its correlation with the Chinese Wechsler Memory Scale-revised (WMS-RC) in patients with memory disorder caused by severe traumatic brain injury (sTBI). The subjects included 20 sTBI patients with memory disorder and 22 healthy individuals. Memory function was measured by using WMS-RC. Behavioral and ERP responses were recorded on-line during performance on a battery of picture recognition and the responses were analyzed off-line for recognition memory effects. Mean memory quotient (MQ) of patients with sTBI was significantly lower than that of the control group. Mean reaction time (RT) was significantly longer and the mean correctness rate (CR) of picture recognition was significantly lower in sTBI group than that of the controls. In controls, the main components of average ERP of picture recognition includes two positive-going waves, designated as P(170) and P(500), that appear 170 ms and 500 ms after stimulation when the subject could later successfully recall and recognize the pictures. P(500) amplitude of target stimulus was significantly higher than that of non-target stimulus. Compared to controls, P(500) responses of sTBI group were significantly delayed in latency (P<0.001) and lower in amplitude (P<0.001). P(500) latency showed significant negative correlation with MQ and the scores of "addition", "visual recognition", "picture recall", "visual reproduction" and "tactile memory" in WMS-RC. ERP of picture recognition provides a neurophysiological approach to directly assess memory impairment, and P(500) may serve as a helpful index for memory disorder caused by sTBI in forensic practice.
Brain Injuries/*complications ; Case-Control Studies ; Evoked Potentials/*physiology ; Memory Disorders/*etiology ; Memory Disorders/*physiopathology ; Pattern Recognition, Physiological/*physiology ; Wechsler Scales ; Young Adult