Objective:To investigate the expressions of CD44v3 and CD54 in oral premalignant epithelia and its significance in the carcinogenesis of oral mucosa.Methods:The expressions of CD44v3 and CD54 in 19 cases of normal mucosa and mucosal hyperplasia,22 case of dysplastic epithelia,26 cases of oral cancer mucosa were detected by using immunohistochemistry.Results:CD44v3 and CD54 proteins showed no expression or micro-expression in normal and hyperplastic epithelia.The expressions increased in the dysplastic epithelia and mucosa of oral cancer gradually.The difference between the expressions of the three groups was significant(P

Objective To probe into the optimal concentration of Wnt-11 to induce the differentiation of bone marrow mesenchymal stem cells (BMMSCs)into cardiomyocyte-like cells in vitro.Methods BMMSCs were isolated from the bone marrow of SD rats using whole bone marrow culture method.After cultured for 48 h, BMMSCs of the second generation were utilized for directed induction.Based on the final concentration of Wnt-11 , BMMSCs were divided up into Group A (100 ng/mL),Group B (200 ng/mL),Group C (400 ng/mL)and Group D (blank control).After 72-hour induction,the cells were cultured in complete medium for 4 weeks while cells in Group D were cultured only in the complete medium.The morphological changes were observed under the phase contrast microscope.Surface antigen expression of BMMSCs was identified by flow cytometry.When cells were cultured for 4 weeks,the expressions of Desmin,Connexin43 and cTnI were detected by immunocytochemistry. Meanwhile, the ultrastructural changes were observed using transmission electron microscope. The mRNA expressions of cardiac transcription factors GATA-4,Nkx2.5 andα-MHC in BMMSCs were detected by RT-qPCR at 1,2 and 4 weeks after induction.Results Primary BMMSCs formed cell colonies at 2 weeks;the cells were mainly fusiform or star-shape,and a few irregularly-shaped ones were also visible.The passaged cells were larger than those of primary culture.After induction,the cells exhibited long shuttle-shape and were aligned in parallel. Flow cytometery displayed that the positive rate of the surface antigens of BMMSCs CD29,CD45,and CD90 was 97.9%,0.4% and 99.5%,respectively.When BMMSCs-induced via Wnt-11 were cultured for 4 weeks,Desmin, cTnI and Connexin43 were all positively expressed in induction groups.Whereas in the blank control group they were slightly positive or negative;the positive rate in Group B was the highest (P<0 .05 ).Transmission electron microscopy exhibited that organelles such as rough endoplasmic reticulum,mitochondria,as well as some ribosomes were visible in the cytoplasm of these cells in each induction group.In addition,myofilaments were arranged in parallel in the cytoplasm.The cells in induction groups could express GATA-4 and Nkx2 .5 in the first week,and then the expression of them decreased in the second week,but then increased in the fourth week;gene expression in induction Group B was significantly higher than in the other two induction groups (P<0 .05 ).The expression of GATA-4 and Nkx2 .5 in Group D was 1 ,α-MHC was not expressed in the four groups during the culture period. Conclusion Wnt-11 can induce the differentiation of BMMSCs into cardiomyocyte-like cells in vitro,and the optimal concentration of Wnt-11 is 200 ng/mL.

In order to explore the pathogenic characters,diagnosis and treatment of canine leptospirosis,a retrospective analysis of the pathogenetic factor,diagnosis and treatment of three dogs were reported which diagnosed as canine leptospirosis in China Agricultural University Teaching Animal Hospital from August to November,2015.The infected dogs showed symptoms of depressed,jaundice,anorexia and vomiting.Two of them had fever history and contacted the water resource outside before getting ill.All three dogs were confirmed anemia,liver and kidney damage by laboratory examination.They were diagnosed with canine leptospirosis according to the positive results of the Leptospira PCR examination base on urine.During the diagnosis and treatment process,one of them was dead,the rest two recovered after antibiotic and symptomatic therapy.In conclusion,dogs which infected with leptospirosis may have a contact with the infected water before.The Leptospira PCR examination base on urine can be used for the diagnosis method of the disease.As to treatment,the use of penicillin and tetracycline drugs such as doxycycline is recommended.

Objective To analyze the morbidity,mortality,and hospitalization of complications in late preterm infants.Meth-ods A total of 389 late preterm infants were retrospectively analyzed.The influence of delivery mode on respiratory complications was compared.And the influence of respiratory complications on length of hospital stay,hospital expenses and mortality were ana-lyzed and compared.Results Our data showed that main complications were diagnosed as respiratory complications(23.65%).Oth-er complications were diagnosed as hypoglycemia (20.57%),feeding intolerance (1 7.48%),hyperbilirubinemia (14.91%),and in-tracranial hemorrhage (8.23%).Respiratory morbidity was significantly higher in preterm infants by caesarean section than by va-ginal delivery (32.34% vs .10.39%,P <0.05).The time and cost of hospitalization and the mortality were higher in late preterm infants accompanied by respiratory complications than those in infants without respiratory complications(P <0.05).Conclusion There were very prone to a variety of complications in late preterm infants.In order to reduce the morbidity of respiratory complica-tions in late preterm infant,indications for caesarean section must be strictly mastered.We should pay more attention to respiratory complications,which might be helpful to short the length of hospital stay,save hospitalization expenses and reduce the mortality.

Objective To investigate the significance of the expression of serum LDH and β2-MG in predicting survival of NHL patients and there correlations with clinical parameters. Methods The serum levels of LDH and β2-MG of 429 patients with NHL were measured by rate method and immunoturbidimetry.Besides,the clinical reference of the patients such as gender,age,B symptoms,clinical stages,extranodal sites and Karnofsky evaluation were summarized and grouped.All patients' survival status were achieved via phone and letter. All statistical analyses were performed using the SPSS program for Windows (version 16.0).Results The levels of LDH and β2-MG in age,B symptom,the advanced stage,extranodal sites≥2,Karnofsky evaluation were obviously higher than their counterpart and the difference was significant (P＜0.001). The median of LDH and β2-MG lever in age group was 229 U/L,190 U/L, 2.4 mg/L,1.7 mg/L. In B symptom was 260 U/L,192 U/L,2.3 mg/L, 1.7 mg/L. In clinical stage was 252 U/L, 175 U/L, 2.5 mg/L, 1.6 mg/L. In extranodal sites group was 278 U/L,195 U/L,2.4 mg/L,1.8 mg/L.In Karnofsky evaluation group was 250 mg/L,195 mg/L,2.2 mg/L,1.8 mg/L.LDH level of invasive lymphoma was significantly higher than that of indolent ones (median 211U/L,175 U/L,P=0.002),but there were no difference in the 32-MG level (P=0.937). There were no statistical significance about LDH and β2-MG levels in gender and cell type (P＞0.05).Overall survival rates were different between the abnormal LDH group and the normal LDH group (χ2=119.029, P＜0.001).Overall survival rates were different between the elevated β2-MG group and the normal β2-MG group (χ2 =104.733,P＜0.001).Overall survival rates of the abnormal LDH/β2-MG group was significantly lower than that of normal group (x2=192.326,P＜0.001).Multivariate analysis in Cox regression showed that LDH,β2-MG,age,clinical stages, extranodal sites, Karnmofsky evaluation and aggressive were independent prognostic factors.Conclusion The level of serum LDH and β2-MG can be taken as an auxiliary clinical index to evaluate the prognosis of the NHL patients.

Objective Studies show that the abnormal ex-pressions of APC and survivin play important roles in the development and progression of colon cancer .Survivin shRNA and APC double gene co-expression lentiviral vector was constructed to observe whether it could be successfully expressed in HT-29 colon cancer cells and whether it could impact on colon cancer cell apoptosis . Methods We selected the best shRNA interference fragment from the construc-tion of three pairs of complementary shRNA fragments and connected it with the effective fragment of APC ( aa1020-1698 ) to construct a double gene co-expression lentiviral vector .HT-29 cells were divid-ed into experimental group , empty loading group and blank group .HT-29 cells were observed by fluorescence microscopy after infec-tion.Survivin and APC expression levels were observed by real time PCR and western blot .Apoptosis was detected by caspase-3 activi-ty assay. Results ①We successfully constructed three pairs of shRNA sequences and proved that they had no human gene homolo -gous to the rest.Real time PCR analysis showed that the best sequence was shRNA 3.②After the sequence alignment of constructed shRNA vectors, three pairs of shRNA sequences were completely the same with the first designed sequence .Green fluorescence was observed in HT-29 cells by fluorescence microscope .The survivin content in experiment group (31.71 ±1.49) was significantly de-creased compared with empty loading group (100 ±0) and blank group(95.12 ±2.15)(P<0.05).The expression level of APC mR-NA was significantly increased compared with empty loading group (0 ±0) and blank group(0.51 ±0.15)(P<0.05).③The relative ratio of apoptosis in experiment group (0.573 ±0.050) was significantly increased compared with empty loading group (0.390 ± 0.040) and blank group(0.407 ±0.030)(P<0.05). Conclusion We have successfully constructed survivin-shRNA-APC double gene co-expression lentiviral vector which can be successfully expressed in HT-29 colon cancer cells , providing references for subse-quent gene therapy by the use of the carrier .

Objective To investigate the effects of the microbubbles combined with different mechanical index ultrasound irradiation on ultrastmcture and migration of colon cancer cells.Methods The experimental study was conducted.Colon cancer cells inn vitro (Lovo ceils) were cultured and divided into 4 groups,ceils in the A group were not treated and cells in the B,C and D groups were treated by microbubbles combined with different mechanical index ultrasound irradiation (mechanical index were 0.20,0.80 and 1.45).The changes of ultrastructure and migration of cells were observed using laser scanning confocal microscope and MilliceIl-PCF cell culturechamber method,respectively.Measurement data with normal distribution were represented as (x)±s.Comparisons among groups were analyzed by the one-way ANOVA.Pairwise comparisons were done by the t test.Results (1)Effects of the microbubbles combined with different mechanical index ultrasound irradiation on ultrastructure of Lovo cells:Lovo cells in the A group showed big nucleus,less plasma,regular arrangement,jagged-like or more irregular varicosity surrounding nucleus,twisted euchmmatin,expansion of nucleus cisternal space,homogeneous distribution and normal development of granular soil and clear mitochondrial ridge-like structures.Lovo cells in the B group showed big nucleus with regular arrangement,obvious nucleolus margination,endoplasmic reticulum dilatation,normal development of mitochondrion and clear mitochondrial ridge-like structures.Lovo cells in the C group showed broadening nucleus space,abnormal nucleus with karyopycnosis,chromatin condensation,a few remaining or obvious dilatation of rough endoplasmic reticulum,typically consisting of different fragments or bubbles,cytoplasmic vacuoles changes and decreasing mitochondrial ridge-like structures.Lovo cells in the D group showed big and irregular nucleus,nucleolus margination,obvious endoplasmic reticulum dilatation,fewer mitochondrion with extended cell area and swelling shape,rare mitochondrial ridge-like structures with disordered or broken arrangement,even disappearing.(2) Effects of the microbubbles combined with different mechanical index ultrasound irradiation on migration of Lovo cells:Millicell-PCF cell culture chamber method showed that number of migration of Lovo cells were respectively 63±7,61±4,21±3 and 19±5 in the A,B,C and D groups,with a statistically significant difference (F=55.040,P＜0.05).There were no statistically significant difference in migration of Lovo cells between group A and B (t =1.571,P＞0.05) and between group C and D (t =2.013,P＞0.05).There were statistically significant differences in migration of Lovo cells between group A and C or D (t=7.861,10.652,P＜0.05) and between group B and group C or D (t=7.161,10.453,P＜0.05).Conclusion Microbubbles combined with high mechanical index ultrasound irradiation can make the ultrastructural alterations in the cancer cells,resulting in tumor cell degeneration and death,ultimately inhibit tumor cell migration and metastasis.

Primary hepatic lymphoma (PHL) is an extremely rare disease without any unified diagnostic criterion.The symptoms are usually nonspecific.Liver biopsy remains the most valuable tool for diagnosis of PHL.The predominant histology of PHL is diffuse large B-cell lymphoma.The therapeutic modalities are variable,including surgery,chemotherapy,radiotherapy,or combination of the various processes.This article described a 33-year-old man with diffuse large B-cell PHL who was treated at the Affiliated Cancer Hospital of Shanxi Medical Univeitity Blood Disease Diagnosis and Treatment Center in February 2014.The patient benefited from eight-cycle chemotherapy.At present,the patient is disease-free and undergoes regular follow-up.

Objective Post-transcription RNA interference (RNAi) is more and more widely applied in multigene therapy.This study aimed to establish an subcutaneous xenotransplanted tumor (SXT) model of human HT-29 colon carcinoma in nude mice and investigate the effects of the survivin shRNA-APC double gene co-expression lentiviral vector on the growth of SXT.Methods Thirty-five nude mice were equally divided into five groups, double-gene survivin shRNA, survivin shRNA, APC, empty vector, and blank, and injected into the left anterior axillary with respective stably transfected cell lines and human HT-29 colon carcinoma cells, all at 2×106/mL, to establish an SXT model of human HT-29 colon carcinoma.The inhibition rate of tumor growth was calculated by measuring the size and weight of the SXT, the expressions of survivin mRNA and protein in the tumor tissue detected by real time PCR and immunohistochemistry respectively, and the apoptosis of the HT-29 colon carcinoma cells determined by TUNEL.Results The mean size and weight of the SXT were significantly reduced in the double-gene survivin shRNA-APC, survivin shRNA, and APC groups as compared with the blank and empty vector groups (P<0.05), though increased in the survivin shRNA and APC groups in comparison with the double-gene group (P<0.05).The expressions of survivin mRNA and protein in the tumor tissue were remarkably lower in the double-gene survivin shRNA-APC, survivin shRNA, and APC groups than in the blank and empty vector groups (P<0.05), even lower in the double-gene group than in the survivin shRNA, and APC groups (P<0.05).The apoptosis rate of the HT-29 colon carcinoma cells was markedly up-regulated in the double-gene survivin shRNA-APC ([56.72±3.17]%), survivin shRNA ([33.64±2.03]%), and APC groups ([31.19±1.79]%) as compared with the blank ([9.89±0.31]%) and empty vector groups ([10.06±0.43]%) (P<0.05), even more significantly in the double-gene than in the survivin shRNA and APC groups (P<0.05).Conclusion The survivin shRNA-APC double gene co-expression lentiviral vector can reduce the expression level the survivin gene, promote the apoptosis of colon carcinoma cells, and suppress the growth of the subcutaneous xenotransplanted tumor.

Objective To explore the potential genetic variants related with late-onset Alzheimer's disease(LOAD),and to broaden the AD genetic susceptibility factor profile.Methods We targetedly sequenced four lipids metabolism related genes including apolipoprotein E(APOE),sortilin-related receptor gene (SORL 1),apolipoprotein C-I (APOC1) and clusterin (CLU),validated these candidate variants in patients with LOAD and mild cognitive impairment (MCI)and the controls with normal cognitive function,and made a bioinformatic analysis.Results The mutation in the allele of CLU rs117389184,a low-frequency single nucleotide polymorphism,would increase the risk of LOAD and MCI[OR (95 ％ CI):3.56 (1.08-4.84),P =0.026;OR (95 ％ CI):3.06 (0.98-4.03),P =0.026],suggesting CLU rs117389184 gene variant was associated with cognitive impairment.This variation could affect CLU gene expression by interfering with the binding of transcription factors.Conclusions CLU rs117389184 is a new genetic risk variant for LOAD and MCI.