Objective: To study the characterization of the cultured cervical cancer cell line transfected with anti- HPV16E6-ribozyme, and to investigate the effect of ribozyme on proliferation and apoptosis of cervical cancer cell. Metliods: Anti-HPV16E6-ribozyme had been designed to cleave the HPV16E6 gene. With the method of lipofectin transfec- tion, the anti-HPVI6E6-ribozyme and empty eucaryotic expressing plasmids were transfected into CaSKi cell, which named as CaSKi-R, CaSKi-P respectively. The amounts of E6 mRNA in the three kinds of cells were detected by northern blot. Cell cycle was detemined by flow cytometry, and cell apoptosis was examined by fluorescent (Hoechst) staining and TUNEL. The expression of some genes, including c-myc, bcl-2, p53, and fas, was also detected by flow cytometry analy- sis. Results: Northern blot showed that E6 mRNA was less in CaSKi-R than in CaSKi. In CaSKi-R cells, cycle was arres- ted in G1 phase, with decreasing in percentage of S phase cells. The apoptosis rate of CaSK1-R cell was much higher than those of CaSKi and CaSK1-P. Anti-HPV16E6-ribozyme could reduce the expression of E6, c-myc, bcl-2 genes on CaSKi- R cells, and increased the expression of p53. While this phenomenon was not found on the CaSK1-P cells. The expression of fas was similar in the three kinds of cells. Conclusion: Anti-HPVE6-rivozyme induces apoptosis of human cervical cancer CaSKi cells. The mechanisms may be the decrease of E6 gene's expression, and the succedent changing of some genes'expression.

OBJECTIVE To probe the risk factors of the secondary lung infection in patients of postoperative(cranio)-(cerebral) trauma as well as the ways to control them.METHODS A prospective and review study was carried out in 718 cases with cranio-cerebral trauma admitted in our hospital for operation from Jan 2002 to Dec 2004.RESULTS Among 718 cases,39(5.43%) developed lung(infection),occupied 65% of the total number of the hospital(infection) in the neurosurgery.After an integrated(total and systemic) treatment,the cure rate was(69.23%(27 cases);) effective: 15.3%(6);improved: 7.7%(3); died: 7.7%(3).CONCLUSIONS Oxygen(inhalation,) nasal(feeding),sputum drawing out,(cannular) tracheotomy,and use of respirator after operation are the causes of the lung infection and it can be controlled effectively if keeping the surroundings clean,keeping strict ness in disinfection-isolation institution,improving the mangement of respiratory tract and the care of oral cavity,proper use of antibiotics,and improving immunity.

In this paper, we investigate the boundedness character, the global attractivity and the periodic nature of the svstem of ratiol difference equatious:xn+1=p+yn-k/xn,yn+1=q+xn-k/yn,n=0,1,2..., where p>0, q>0, k∈ of the system of rational difference equations:xn+1=P+yn-k/xn, yn=q+xn-k/yn,n=0,1,2 {1,2,…} and the initial values x1, y1∈(0,∞), i=- k, -k+1,… 0. Some new results are obtained.

Objective: To investigate the characteristics of the cultured cervical cancer cell line transfected with anti HPV16E6 ribozyme, and to investigate the possibility and practicality of ribozyme in treatment of cervical cancer. Methods: The anti HPV16E6 ribozyme and empty eucaryotic expressing plasmids were transfected by lipofectin transfection into CaSKi cell, which named as CaSKi R, CaSKi P respectively. The morphology and the soft agar forming ability were studied. The expression of E6, PCNA and C erbB 2 genes was studied through Flow Cytometry. The tumorgenicity of each cell was detected by injecting cells into the nude mice skin. Three groups of nude mice were injected by CaSKi, CaSKi R and CaSKi P cell separately. Another group of mice was injected by CaSKi cell on right side and CaSKi R cell on left side. Results: There is no distinct difference of the morphology and growth rate between CaSKi and CaSKi P, but the growth rate of CaSKi R decreased. The soft agar forming rate of CaSKi P was similar with that of CaSKi cells, while that of CaSKi R was found decreased. The result of flow cytometric analysis showed that anti HPV16E6 ribozyme could reduce the expression of E6, PCNA and C erbB 2 genes on CaSKi R cells, while this phenomenon was not found on the CaSKi P cells. The tumorgenicity of CaSKi R in nude mice was decreased compared with CaSKi and CaSKi P cells. Conclusion: Anti HPVE6 rivozyme could partly reverse the malignant phenotypes of CaSKi cells. The reason may be the decrease of E6 gene expression, and the succeeding decrease of the PCNA and C erbB 2 genes′ expression.

Adjuvant treatment, which includes chemotherapy and endocrine therapy, for early breast cancer may impair bone den-sity, resulting in bone loss. The third generation bisphosphonate-zoledronic is an anti-resorptive agent that inhibits osteoclast-mediated bone resorption. This drug can be mainly used in the treatment of hypercalcemia caused by bone metastases of the cancer. The Zome-ta-Femara Adjuvant Synergy Trial revealed that immediate zoledronic acid and endocrine therapy not only prevents bone loss, but also reduces recurrence. The Austrian Breast and Colorectal Cancer Study Group-12 (ABCSG-12) also confirmed that zoledronic acid, when combined with endocrine therapy, could reduce the risk of cancer disease progression and death. Moreover, preclinical studies and clini-cal trials have demonstrated the synergistic antitumor effects of chemotherapy and zoledronic acid. Neo-adjuvant zoledronic acid to re-duce recurrence trials showed that the addition of zoledronic acid to adjuvant chemotherapy significantly reduced the risk of disease pro-gression and death in postmenopausal women more than five years postmenopause at the beginning of the study or over 60 years of age at the baseline. The ABCSG-12 subgroup analysis based on age (≤40 years or>40 years) also showed that zoledronic acid can signifi-cantly improve the prognosis in women who were over 40 years at the study entry. These results suggest that zoledronic acid administra-tion in patients with lowered estrogen levels (naturally or as a consequence of adjuvant treatment) easily exerts anti-tumor effects. How-ever, the optimal dose and duration of zoledronic acid requires further studies. More clinical trials should be performed to provide suffi-cient evidence to support the effectiveness of zoledronic acid in the treatment of early breast cancer.

BACKGROUND:Tissue engineering requires a lot of seed cells. Osteoblasts have become important seed cells in bone tissue engineering. However, it is difficult to culture the osteoblasts, and cellnumber, purity, proliferation and differentiation activity are different obtained by different culture methods. OBJECTIVE:To identify and compare three common primary osteoblat culture methods, and to explore a method for the primary culture of osteoblasts which is easy to operate, economical and effective, in order to provide basis for the further experimental research. METHODS:Calvarias were dissected from newborn Sprague Dawley rats in 72 hours, and osteoblasts were isolated with col agenase digestion method, sequential digestion method and bone tissue method respectively. The morphological observation and cytochemical staining were performed, the growth curve of the cells was drawn with cellCounting Kit-8 method, and the rate of living osteobalsts was counted with trypan blue staining. RESULTS AND CONCLUSION:The proliferation of the insolated and cultured osteoblasts was wel with typical characteristics of osteoblasts, cytochemical staining results were positive. Compared with the sequential col agenase digestion method, the col agenase digestion method presented higher production of osteoblasts and higher cellsurvival rate (P<0.05), and the col agenase digestion method was easier than the sequential col agenase digestion method and cost less than sequential col agenase digestion method. Bone tissue method was the easiest method with less damage to cells, but bone tissue method presented lower production of osteoblasts and cost much more time, which cannot be used in large-scale osteoblast culture. The col agenase digestion method is a simple, efficient and ideal method for isolation and culture of primary osteoblasts.

Objective: To investigate the effects of HPV16E6-ribozyme on telomerase activity in cervical carcinoma cell line CaSKi and the related mechanisms. Methods: Anti-HPV16E6-ribozyme and blank eucaryotic plasmids were transfected into CaSKi cells via lipofectin, and the resultant cells were named as CaSKi-R and CaSKi-P, respectively. The expression of ribozyme in transfected cells was observed by RNA dot blotting. The expression of E6 mRNA and protein in the 3 kinds of cells were detected by Northern blotting and Western blotting, respectively. Telomerase activity was determined by TRAP-Elisa method; the expression of P53, c-myc, hTERT and hRT mRNA were examined by RT-PCR.Results: RNA dot blotting showed that anti-HPV16E6-ribozyme was stably expressed in transfected CaSKi-R cells. Western blotting showed that the expression of E6 mRNA and protein in CaSKi-R cells was obviously lower than that in CaSKi and CaSKi-P cells. The telomerase activities in CaSKi,CaSKi-P and CaSKi-R cells were (0.89?0.14), (0.90?0.11) and(0.36?0.06),respectively. The inhibitory rate of telomerase activity in CaSKi-R cells was 59.55%, which was significantly lower than those in CaSKi and CaSKi-P cells (P

Objective To investigate the prognosis and its influencing factors of post-encephalitic epilepsy ( PEE) .Methods Fifty viral encephalitis patients with PEE were followed up, which prognosis was observed by whether their no seizures for at least 1 year or not.The clinical data of patients was collected, and the relate risk factors on the poor prognosis of PEE were analyzed.Results The disturbance of consciousness and the seizure presence during acute encephalitis, more than ten seizures in poor prognosis group were much higner than the good one (all P<0.01).The age, sex, seizure type, dysphrenia and neurological defect in acute encephalitis, status epileptisus in acute encephalitis, abnormal EEG and abnormal head CT/MRI had no correlation with poor prognosis ( all P>0.05 ) .Multiple logistic regression analysis demonstrated that disturbance of consciousness during acute encephalitis、seizures presence in acute encephalitis were independent risk factors of poor outcome in PEE ( OR=7.269,95%CI:1.22 -43.35,P=0.029;OR =22.893,95%CI:4.02 -130.43,P=0.000).Conclusion Disturbance of consciousness during acute encephalitis, seizure presence in acute encephalitis both are the single risk factors of poor prognosis in PEE.

Objective To investigate the prognostic values of neutrophil to lymphocyte ratio (NLR) and platelet count before and after chemotherapy in patients with advanced non-small cell lung cancer (NSCLC).Methods From October 2011 to December 2012,70 patients with advanced NSCLC in Anyang Cancer Hospital of Henan Province were collected.NLR and platelet were calculated from complete blood counts in laboratory test before and after 2 cycles of chemotherapy.The patients were divided into two groups according to the NLR and platelet count:low NLR group (≤3.43)and high NLR group (>3.43);normal group (100 ×109 /L