Objective To study the expression and significance of human tissue kallikrein gene 6(KLK6) in cervical cancer tissues.Methods With glyceraldehyde 3-phosphate dehydrogenase (GAPDH)as reference,the expression of KLK6 in 80 cases of cervical cancer tissues (40 cases with metastasis and 40cases without metastasis) and 40 cases of normal cervical tissues was determined by Taqman probe real-time quantitative reverse transcription-polymerase chain reaction,and analyzed the relationship between cervical cancer occurring and KLK6 expression with clinical data and pathological dats.Results The expression of KLK6 in normal cervical tissues[(1.06 ± 0.40) × 10-3] was lower than that in cervical cancer tissues without and with metastasis[(4.41 ± 1.70) × 10-3,(32.22 ± 6.70) × 10-3],and there was significant difference (P＜0.01).The expression of KLK6 in Ⅰ a,Ⅰ b,Ⅱ a stage of cervical cancer tissues with metastasis was (30.42 ± 5.00) × 10-3,(31.64 ± 1.30) × 10-3,(33.02 ± 8.00) × 10-3,and there was no significant difference among them (P ＞ 0.05).The expression of KLK6 in Ⅰ a,Ⅰ b,Ⅱ a stage of cervical cancer tissues without metastasis was (4.12 ± 1.10) × 10-3,(4.35 ± 1.30) × 10-3,(4.82 ± 1.90) × 10-3,and there was no significant difference among them (P＞0.05).There was significant difference in the expreesion of KLK6 in Ⅰ a,Ⅰ b,Ⅱ a stage between cervical cancer tissues with metastasis and cervical cancer tissues without metastasis (P ＜0.01).Conclusion KLK6 can stimulate the cervical cancer cell proliferation,and participate in the progresses of cervical cancer metastasis.

Objective To explore the association of expression of apoptosis-associated gene BAK and cFLIP with the biological behaviors in endometriosis. Methods The expression of BAK and cFLIP protein gene in eutopic and ectopic tissue samples from 40 cases with pathologically confirmed ovarian endometriosis and 40 cases with pathologically confirmed normal endometrium was detected by immunohistochemical method. Results ①The expression of BAK and cFLIP protein gene was found in three groups of different endometrial tissue. ②The expression of BAK protein gene was increased gradually in ectopic endometrial , eutopic endometrium and normal tissue and there was significantly difference between every two groups ,while cFLIP was contrary expressed (P <0.05); ③In normal endometrium, The expression of BAK protein gene in secretory phase was higher than that in proliferative phase (P < 0.05), while cFLIP was contrary expressed, in eutopic endometrium and ectopic endometrial tissue , The expression of BAK and cFLIP protein gene in secretive phase showed no statistically significant difierence with that in proliferative phase (P > 0.05). ④The expression of BAK protein gene in severe group (Ⅲ-Ⅳ period) is lower than mild group both in eutopic or ectopic endometrial tissues,while cFLIP was contrary expressed (P < 0.05). ⑤The expression of BAK and cFLIP was negatively correlated with each other in ectopic endometrium (r=-0.389,P< 0.05). Conclusion BAK and cFLIP was negatively expressed in EMS, which may take a part in the endometrial apoptosis and disorderly proliferation. BAK and cFLIP may play an important role in the the diagnosis and treatment of endometriosis.

Objective To investigate the influence of Chinese medicine ( CM) treatment on life span of middle-late primary hepatic carcinoma (PHC) patients. Results A multicenter retrospective cohort study was carried out in 489 PHC patients at the stages of Ⅱb, Ⅲa and Ⅲb collected from 15 domestic hospitals. With CM treatment as the exposure factor, the enrolled patients were divided into CM group, western medicine (WM) group and the CM-WM combination group. The main outcomes included median survival time (MST) and survival rate, and the effects of CM treatment and minimally invasive therapy on MST were observed. Results Half-a-year survival rate, one-year survival rate and two-year survival rate were 50%, 9%, 1% in CM group, 70%, 30%, 6% in CM-WM combination group, and 50%, 10%, 0% in WM group, respectively. The survival rates in CM-WM combination group differed from those in CM group and WM group (P<0.01), but the differences of survival rates between CM group and WM group were insignificant (P>0.05) . MST at the stages of Ⅱb, Ⅲa, Ⅲb in CM-WM group was obviously prolonged as compared with the other two groups (P<0.01), and MST at the stages of Ⅲa, Ⅲb in CM group was also obviously prolonged as compared with WM group (P<0.05) . The difference of MST at the stage of Ⅱb was insignificant between CM group and WM group ( P>0.05) . CM treatment and minimally invasive therapy were effective on prolonging the survival time of PHC patients ( P<0.01) . The results of COX regressionanalysis showed that Karnofsky scores, CM treatment and minimally invasive therapy were the preventive factors for the prognosis. Conclusion CM-WM combination group has the best long -term therapeutic effect. CM -WM combination treatment is effective on increasing MST and long-term survival rate, in particular for PHC patients at the stages ofⅡb, Ⅲa and Ⅲb. CM treatment and minimally invasive therapy are helpful for the prolongation of the survival time of PHC patients.

Objective To observe whether appropriate intermittent exercise at the ischemic threshold can safely promote collateral circulation in an ischemic area of the myocardium through the increased expression of vascular endothelial growth factor(VEGF)and its receptor fetal liver kinase-1(Fik-1). Methods A balloon constrictor was surgically implanted in the first obtuse marginal coronary artery(OM1)of miniature pigs.The subjects were divided into 3 groups:a sham-operation group,a pure ischemia group,and an exercise training group.Subjects in the exercise training group performed individualized treadmill programs 30 min daily,5 d per week,for 8 weeks,including 2 two-minute episodes of exercise-induced ischemia.Two pre-exercise episodes of pure ischemia induced by brief OM1 occlusion were also conducted.Only pure ischemia was induced in the pure ischemia group,and the sham-operation group remained sedentary for the experimental period.Relative myocardial blood flow(RMBF)was measured using microspheres.VEGF and Flk-1 expression levels were measured by Western blotting and real time RT-PCR analyses.Cardiac troponin I(ctnI)levels were determined using an enzyme-linked immunosorbent assay(ELISA). Light and electron microscopy were employed to examine myocardia damage in the ischemic area.Results RMBFs in the exercise training group were significantly higher than those in the pure ischemia and sham-operation groups. RMBFs in the pure ischemia group were significantly higher than those in the sham-operation group.The expression of VEGF and Flk-1 proteins and mRNAs in the exercise training group were significantly higher than those in the pure ischemia and sham-operation groups,and the levels in the pure ischemia group were also significantly higher than those in the sham-operation group.After training,no myocardial damage and no ctnI increase was observed in the pure ischemia group.Microscopy revealed no obvious structural changes. Conclusion Intermittent exercise at the isehemia threshold intension can safely promote coronary collateral formation through upregulation of VEGF and Flk-1 expression in the ischemic myocardial area of a porcine model.

Objective:Detecting nasopharyngeal carcinoma(NPC) patients′olfactory before and after radiotherapy for investigating the infection of radiotherapy on these patients′olfactory.Method:100 NPC patients′olfactory were detected before the start of radiotherapy and after the termination of therapy in 3,6,12,24 and 36 months.Result:Patients′smell acuity decreased sharply 3 months after therapy,but varying degrees of recovery were noted in 6 and 12 months. However their smell acuity became worse in the following time.Conclusion:NPC patients′smell acuity can be profoundly affected by radiotherapy.And after radiotherapy, their smell acuity will decrease wave-likely and cannot fully recover.

OBJECTIVE:To provide reference for rational drug use in the clinic. METHODS:The cases of disulfiram-like reac-tion collected from the First Affiliated Hospital of Guangzhou University of TCM from July 1st in 2012 to June 30st in 2014,were analyzed and compared retrospectively. The epidemiological characteristics,causes,clinical characteristics and treatment methods were summarized. RESULTS:Disulfiram-like reaction mainly occurred in male(86.59%),mainly in young and middle-aged men aged 21-50 years old (78.05%). It might be induced by using cephalosporin antibiotics (as cefoperazone,cefperazone/sulbactam, etc.),nitroimidazole (as metronidazole,tinidazole,etc.),drinking or contacting beverage,food or drug containing ethanol. This reaction involved the skin and its accessories system,circulatory system,digestive system,nervous system,respiratory system, etc. Its clinical manifestations mainly were skin flush,chest tightness,shortness of breath,palpitation,nausea,vomiting,dizzi-ness,headache,etc.,occasional confusion,drowsiness,xerostomia,etc. Response to the severe disulfiram-like reaction,the foun-dation treatment combined with symptomatic treatment showed a good effect with remission of symptoms in 1 to 3 hours and no deaths. CONCLUSIONS:When the drugs with mercapto-1-methyltetrazole perssad as cephalosporin antibiotics,nitro imidazoles, etc.,are used,we should avoid drinking or taking drugs or food containing ethanol,to prevent the occurrence of disulfiram-like re-action.

BACKGROUNDSepticemia and inflammation-mediated septic shock caused by Vibrio vulnificus (VV) is strongly associated with chronic liver disease. This study examined the effects of antimicrobial therapy on expression of hepatic toll-like receptors and inflammatory cytokines in rats with alcohol-induced liver disease complicated by VV sepsis.

METHODSMale Sprague-Dawley rats were assigned to the following treatment groups: normal control (N), alcoholic liver disease control (A), antimicrobial-treated alcoholic liver disease control (AA), alcoholic liver disease with VV sepsis (AV), and antimicrobial-treated alcoholic liver disease with VV sepsis (AVA). Alcohol-induced liver disease was observed in all groups except N. Expression of mRNAs encoding hepatic toll-like receptors 2 and 4, myeloid differentiation protein-2, tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-6 and IL-10 was determined by RT-PCR.

RESULTSmRNAs encoding toll-like receptors 2 and 4 and myeloid differentiation protein-2 were significantly up-regulated in group AV as compared to control groups at 2 - 24 hours of sepsis; peak expression occurred at 12 hours. These mRNAs were also up-regulated in group AVA but to lesser degrees than in group AV at comparable time post-infection. mRNAs encoding TNF-alpha, IL-1beta and IL-6 were significantly elevated in group AV as a function of infection. In group AVA as compared to AV, expression of TNF-alpha and IL-1beta mRNAs was lower at 12 - 24 hours post-infection and expression of IL-6 mRNA was lower at 24 hours post-infection. Compared with control groups, IL-10 mRNA expression in group AV was markedly higher at 12 - 24 hours of sepsis. Expression of IL-10 mRNA was lower in group AVA as compared to AV at 24 hours of sepsis.

CONCLUSIONSAntimicrobial therapy reduces expression of toll-like receptors and cytokines in rats with alcohol-induced liver disease complicated by VV sepsis. Monitoring hepatic toll-like receptor and cytokine expression during antibiotic therapy may be valuable for determining the course of VV sepsis in subjects with liver disease.

Adaptor Proteins, Signal Transducing ; genetics ; Animals ; Anti-Infective Agents ; therapeutic use ; Cytokines ; genetics ; Interleukin-10 ; genetics ; Interleukin-1beta ; genetics ; Interleukin-6 ; genetics ; Liver ; drug effects ; metabolism ; Liver Diseases, Alcoholic ; drug therapy ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Sepsis ; drug therapy ; genetics ; microbiology ; Toll-Like Receptor 2 ; genetics ; Toll-Like Receptor 4 ; genetics ; Toll-Like Receptors ; genetics ; Tumor Necrosis Factor-alpha ; genetics ; Vibrio Infections ; drug therapy ; Vibrio vulnificus ; physiology

Objective To explore the perinatal outcomes of women with pulmonary hypertension complicating congenital heart disease(CHD).Methods Clinical data of 45 cases of pregnant women with pulmonary hypertension complicating CHD from Apr 1995 to May 2007 were analyzed and they were divided into three groups:29 cases of slight group[pulmonary hypertension of 30 mm Hg(1 mm Hg=0.133 kPa) to 49 mm Hg],8 cases of moderate group(pulmonary hypertension of 50 mm Hg to 79 mm Hg)and 8 cases of severe group(pulmonary hypertension equal to or higher than 80 mm Hg).The types of CHD,cardiac functional status(New York heart association,NYHA),gestational weeks of pregnancy termination,mode of delivery,pregnancy after CHD operation and outcomes of infants were compared between the groups. Results(1)The highest incidence of CHD were atrial septal defect and ventricular septal defect(58%, 26/45).The rate of pregnant women after CHD operation was 29%(13/45),they were mainly in slight group and their NYHA class were in Ⅰ-Ⅱ.(2)The occurrence rate of NYHA class Ⅲ-Ⅳ was 7/8 in severe group.The rate of NYHA class Ⅰ-Ⅱ as 6/8 in moderate group.The rate of NYHA class Ⅰ- was 97%(28 /29)in slight group.(3)The rate of term delivery was 93%(27/29),preterm labor 3% (1/29),abortion 3%(1/29),and the birth weight was(3153?399)g on average in slight group.The rate of term delivery was 5/8,preterm labor occurred in 3 cases in moderate group.The rate of term delivery was 5/8,preterm labor occurred in 2 cases,and iatrogenic abortion in 1 case in severe group.The average birth weight between slight group and moderate or severe group had a significant difference.(4)Caesarean section rate was 78 %(35/45)among all patients.The rate of cesarean section delivery was 76%(22/29)in slight group,6/8 in moderate group,and 7/8 in severe group.(5)The rate of pregnant women who had portent heart failure or heart failure was 24%(11/45),overall maternal mortality was 4%(2/45).Conclusions The higher the pulmonary hypertension,the worse the outcome of the mother and fetus;The pregnant women with good heart function after cardiac operation would have a good perinatal outcome.Cesarean section is more suitable for those women.

OBJECTIVETo investigate the transcriptional activity of P53 in gastric cancer.

METHODSThe activity of p53 in gastric cancer was investigated by dual-luciferase reporter assay. The coding sequence of the p53 was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and sequenced. The expression of P21WAF1/Cip1, Gadd45alpha and Mdm2 was detected by immunohistochemistry in 76 samples of gastric carcinoma, and their adjacent tissues were analyzed as control.

RESULTSThe p53 activity was higher in human normal cell lines than that of gastric cancer. Furthermore, the transcriptional activity of P53 was lowest in MKN28 cells in which p53 was mutated. The expression level of P21WAF1/Cip1, Gadd45alpha and Mdm2 in the adjacent tissues was higher than that of cancer tissues (P<0.05), and there was a tendency of decline in the positive ratio with the poor differentiation of the carcinoma (P<0.05). In addition, a strong linear correlation was observed between P21WAF1/Cip1, Gadd45alpha and Mdm2 (P<0.05).

CONCLUSIONChanges of P53 transcriptional activity play an important role in the development of gastric cancer. p53 mutation could affect its transcriptional activity. P21WAF1/Cip1, Gadd45alpha and Mdm2 are a group of effecters that could reflect P53 transcriptional activity when detected together in cancer tissues.

Adult ; Aged ; Carcinoma ; genetics ; metabolism ; pathology ; Cell Cycle Proteins ; Cell Line ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Nuclear Proteins ; Proto-Oncogene Proteins c-mdm2 ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transcriptional Activation ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Young Adult

OBJECTIVETo investigate the expression profiles of myocardin gene during the differentiation of bone marrow-derived mesenchymal stem cell to smooth muscle cells in the conditional medium combined with a high concentration of fetal bovine serum (FBS).

METHODSMarrow-derived mesenchymal stem cells were isolated and purified from mouse femoral bone and shinbones using differential adherent methods. Cells at the third passage were induced by 20% FBS in conditioned medium, conditioned medium alone, 20% FBS or 10% FBS alone respectively. Mouse aortic smooth muscle cells were cultured as the positive control. Levels of mRNA and protein expression of myocardin and several smooth muscle cells marker genes were determined by immunofluorescence, RT-PCR and Western blot before and 3, 7, 10, 14 d after the induction. The presence of smooth muscle myofilaments was detected by using transmission electron microscope.

RESULTSNaive bone marrow-derived mesenchymal stem cells displayed multiple morphological forms including fusiform, polygon, oval, and micro-spherical, as compared to the single macro-spindle form after the induction. Typical appearance of peak valley was displayed on the 21st day after induction. At the same time, the expression of smooth muscle marker genes was reinforced along with an up-regulation of myocardin expression. Immunofluorescence study showed that the cells expressing myocardin and smooth muscle marker genes such as alpha-SMA and SM-MHC increased. Fluorescence domain of myocardin translocated from cytoplasm to nucleus and the amounts of double positive cells for myocardin with alpha-SMA or SM-MHC also increased. RT-PCR confirmed that the mRNA expression of myocardin increased gradually and remained stabilized after achieving its peak on the 7th day after induction. The expression of smooth muscle marker genes, alpha-SMA and SM22alpha, remained stable on the 10th day of induction. It was also confirmed by Western blot that the protein expression of both myocardin and alpha-SMA were markedly increased during the induction. Finally, transmission electron microscopy revealed the presence of myofilament on the 21st day after induction.

CONCLUSIONSBone marrow-derived mesenchymal stem cells can be effectively induced into smooth muscle-like cells by conditioned medium combined with 20% FBS. Myocardin plays an important role in the differentiation process of bone marrow-derived mesenchymal stem cells to the peripheral smooth muscle cells.

Animals ; Bone Marrow Cells ; cytology ; physiology ; Cattle ; Cell Differentiation ; physiology ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; physiology ; Nuclear Proteins ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics ; metabolism ; physiology ; Up-Regulation