Objective To establish a radiation-resistant cell subline from a human small cell lung cancer ( SCLC) cell line NCI-H446, providing a pairing research tool for investigating mechanism of radiation tolerance and the reverse strategy in lung cancer .Methods The NCI-H446 cell was radiated repeatedly by increased dose of radiation gradually (total 7500cGy) and a radiation-resistant cell substrain was induced and selected from the survival of cells .The doubling time and cell cycle distribution of the substrain were detected by ATP kit and flow cytometry Assay respectively ;Radiation biology parameters were calculated and analyzed by cell survival curve fitting from multi -target model, SF=1-(1-e-D/D0) N.Results Comparing with the control , The resistant substrain radiobiology parameter values were D 0 ( 1.9673, 2.2756), N(1.0016,2.6008), Dq (0.6783,1.6860)and SF2(0.3623,0.7134) respectively.Cell morphology is more slender and has more tentacles .The SF2 value of radiation-resistant subline is 1.97 times more than that of wild cell line . The proportion of radiation-resistant cells in G2/M-phase was down to 7.84%, compared with the 18.52%of wild cells. Conclusions A radiation-resistant SCLC subline NCI-H446-R is established and may be a useful tool for studying resistant to radiation of SCLC in the future .

Objective To evaluate the diagnostic and prognostic value of BRAF V600E mutation screening of ultrasound-guided fine-needle aspiration (FNA)specimens in patients with thyroid nodule. Methods The BRAF V600E mutation status were assessed in FNA specimens of 104 patients with thyroid nodules before operations.The BRAF mutation status,clinical,and pathology records of the patients were reviewed and the associations between these characteristics and papillary thyroid cancer (PTC ) were analyzed.Results Seventy-one PTC and 14 benign thyroid nodules were included in this study.BRAF V600E mutations were found in 57/71 (80%)PTC.All benign thyroid nodules had no BRAF V600E mutation.The sensitivity,specificity,positive predictive value and negative predictive value of BRAF V600E mutations in differentiation between PTC and benign thyroid nodules were 80%,100%,100% and 50%(P < 0.001 ).In 44 patients with PTC who underwent surgery,the central compartment lymph node metastases and extrathyroidal invasion were not significantly different between BRAF-positive and BRAF-negative PTC (P = 0.283 and 0.307 ).Conclusions BRAF V600E mutation may be a potential tool to facilitate ultrasound in diagnosis of PTC.In patients with PTC,the presence of the BRAFV600E mutation was not significantly associated with prognostic factors.

Objective To compare the clinical outcomes between conventional percutaneous vertebroplasty (PVP) and targeted PVP in the treatment of osteoporotic vertebral compression fractures (OVCFs).Methods A retrospective cohort study was designed to review 215 cases of single level OVCFs hospitalized between January 2014 and December 2015.According to the procedure techniques,the patients were assigned to targeted PVP group (89 cases) and conventional PVP group (126 cases) which was further divided into sufficient filled subgroup (110 cases) and insufficient filled subgroup (16 cases) on basis of cement distribution.Key techniques of targeted PVP included accurate needle insertion to fractured area and cement injection using a push rob with a side opening.Operating time,cement injection volume,rate and types of cement leakage,cement distribution in the fractured area and visual analogue score (VAS) of back pain were compared between the two groups.Results Operating time in targeted PVP group was longer than that in conventional PVP group (P ＜ 0.05).There were no significant differences in cement injection volume and rate and types of cement leakage between the two groups (P ＞ 0.05).None in targeted PVP group showed insufficient cement distribution in fractured area,while 16 cases (12.7％) in conventional PVP group (P ＜ 0.05).No significant differences in preoperative VAS of back pain existed among targeted PVP group,sufficient subgroup and insufficient subgroup (P ＞ 0.05).VAS of back pain was significantly decreased after PVP in three groups (P ＜ 0.05).Difference in postoperative VAS of back pain between targeted PVP group and sufficient filled subgroup was insignificant (P ＞0.05).However,postoperative VAS of back pain in insufficient filled subgroup was significantly increased compared with targeted PVP group and sufficient filled subgroup (P ＜ 0.05).Conclusion Targeted PVP provides sufficient cement to fill the fractured area and decreases incidence of unsatisfactory clinical outcome compared with traditional PVP,indicating a secure and effective new technique in the treatment of OVCFs.

BACKGROUND: It is a hotspot that calcium phosphate and calcium sulphate as the main ingredients are combined with one or more other materials to improve or increase the performance of bone tissue engineering scaffolds. OBJECTIVE: To introduce the research advance of these two kinds of scaffolds in bone tissue engineering. METHODS: The articles related to the bone tissue engineering published during January 2000 to June 2015 were retrieved from CNKI and PubMed databases by computer. The key words were “bone tissue engineering, scaffold, calcium phosphate, calcium sulphate, vascularization” in Chinese and English, respectively. ESULTS AND CONCLUSION: Calcium phosphate and calcium sulfate are characterized as having good biocompatibility, biodegradability, osteoconductivity and complete bone substitutability. However, single use of calcium phosphate or calcium sulfate scaffold has certain disadvantages, both of which are difficult to ful y meet the requirements of the bone defect repair. Improvement can be acquired in the mechanical strength, injectability and biodegradability, as wel as drug-loading and pro-angiogenesis of the scaffold in combination with other materials. In the basal and clinical research, we should explore and develop ideal scaffolds in on the basis of therapeutic aim. However, most of the scaffold studies are stil at the extracorporeal and animal experiment stage, and the comparative studies on composite scaffolds and optimal proportion of those composite scaffolds stil need to be further investigated.

Objective To predict clinical chemotherapy sensitivity of primary ovarian cancer by jointing adenosine triphosphate(ATP) - tumor chemo-sensitivity assay(TCA) method in vitro and detection of drug resistance genes, provide reference for clinical treatment. Methods Forty-seven primary epithelial ovarian tumor samples were collected from the patients who received cytoreductive surgery. Viable ovarian cancer cells obtained from malignant tissue were tested for their sensitivity to carboplatin (CBP), cisplatin (DDP), paclitaxel(PTX) and CBP + PTX using ATP-TCA method in vitro; at same time, real-time quantitative PCR was used to analysis BRCA1 and ERCC1 mRNA relative expression in forty-six specimens (1 frozen tumor samples mRNA were not detected due to serious degradation). The relationship between ATP-TCA test results, clinical indicators, and the effectiveness of the joint prediction on clinical chemosensitivity by combining these two methods were statistically analyzed using chi-square test. Results (1)The results showns that three programs of DDP,CBP and PTX + CBP were significantly related with clinical results(P<0.05) in vitro, in which the compliance rate in PTX + CBP program was the highest 83%(39/47) ,and the predictive sensitivity, predictive specificity, positive predictive value, negative predictive value and predictive accurate rate were 90%,71%,84% and 80% ,respectively.PTX + CBP combined in vitro test results was also related with residual tumor size and neoadjuvant chemotherapy, which was more prone to drug resistance with residual tumor larger than 2 cm (P = 0. 023) and with neoadjuvant chemotherapy (P = 0.011). (2) BRCA1 mRNA expression levels in the clinical-resistant group and the clinical-sensitive group was 0.673 ± 2.143 and - 1.436 ± 2.594 (P=0.008), ERCC1 mRNA expression levels in the clinical-resistant group and the clinical-sensitive group was -0.529 ± 1.982 and - 3.188 ±2.601 (P =0.001). There were also significant correlation among the expression levels of BRCA1 ,ERCC1 mRNA and clinical efficacy (P<0.01). (3)ATP-TCA and detection of drug resistance genes combined to predict the clinical application of PTX + CBP resistance may occur in 8/9 cases. Conclusions ATP-TCA may be an ideal method of in vitro drug sensitivity testing method, which could effectively predict clinical chemotherapy sensitivity. Combination of the drug-resistant associated genes detection method and the ATP-TCA method can increase the predictive effectiveness of ovarian cancer chemosensitivity and guide individual chemotherapy of ovarian cancer.

Objective To investigate the heterogeneity of chemosensitivity in colorectal cancer using an ATP-tumor chemosensitivity assay (ATP-TCA) and the feasibility of individual chemotherapy.Methods An ATP-TCA were used to determine the effect of 16 single or combined cytotoxic drugs in surgical specimens from 50 patients with colorectal cancer.Results There were considerable differences in chemosensitivity between individuals.The most active single drugs in the assay was identified as Navelbine, Hydroxycamptothecin, 5-Fluorouracil and Paclitaxel; 34.1%, 31.6%, 27.6% and 24.3% of specimens showed sensitivity to them, respectively.5-Fluorouracil+Mitomycin+Aytarabine was found to be the most effective combination, for 100% (11/11)specimens were sensitive to this regimen.5-Fluorouracil+Cisplatin+Adriamycin and Gemcitabine+Cisplatin were moderately active regimens.Conclusions There was the heterogeneity of the in vitro chemosensitivity in colorectal cancer.The use of the ATP-TCA provides a method of selecting appropriate anti-cancer drugs in colorectal cancer.

Objective: To study the biological effects of the HPV16Z-Hsp65-E6/E7 fusion protein vaccine on the tumor associated with HPV16 infection. Methods: We tested the cellular immune responsive intensity to the vaccine by the lymphocyte proliferation and CTL response, and studied the therapeutic effect of the vaccine on mouse TC-1 cell transplanted cancer in vivo and the influence on mouse lifetime. Results: The spleen lymphocytes from the C57BL/6 mouse immunized by the Hsp65-E6/E7 vaccine could proliferate obviously in the presence of the protein and TC-1 tumor cell could be lysed specifically by immune activated lymphocytes in vitro. This animal therapeutic experiment in vivo showed that the vaccine suppressed the growth of TC-1 cell transplanted tumor remarkably. Conclusion: The recombined vaccine can induce specific cellular immune response in vitro and suppress HPV16 positive TC-1 tumor cell growth obviously in vivo.

Objective: To develop a therapeutic adjuvantfree protein vaccine against HPV16 which is closely related to cervical cancer of China. Method: First the E6/E7 gene by PCR technology from HPV16z virus strain was isolated in the highrisk cervical cancer area of Shanxi province of China in1990s, and again got the gene segment of Hsp65 from BCG by the same method, mutated the transforming codes in sequences of HPV 16 E6/E7 genes and thus constructed the expression vector pET28aHsp65E6/E7, expressed the Hsp65E6/E7 fusion protein in E.coli BL21(DE3) strain and researched optimal protein purification procedures. Results: The expression vector pET28aHsp65E6/E7 was constructed successfully and E6/E7 gene was mutated correctly. Hsp65E6/E7 fusion protein was renatured and purified on the affinity chromatography column simultaneously. The protein purity achieved 95% after the anionic exchange chromatograph purification. conclusions: This research laid a foundation for further functional study of the therapeutic adjuvantfree protein vaccine——Hsp65E6/E7.

OBJECTIVE:To observe the clinical efficacy and safety of Zidan yinxie granules combined with Acitretin capsule and Compound flumetasone ointment in the treatment of psoriasis vulgaris. METHODS:A total of 97 patients with psoriasis vulgaris were divided into control group(48 cases)and observation group(49 cases)according to number random method. Control group was given Acitretin capsule 10 mg,tid,for consecutive 8 weeks,and then 10 mg,bid,for consecutive 4 weeks and Compound flumetasone ointment for external use,bid. Observation group was given Zidan yinxie granule 4 g,tid,on the basis of control group. Treatment courses of 2 groups lasted for 12 weeks. Clinical efficacy was observed in 2 groups. Psoriasis area and severity index(PASI)score,VAS score,dermatology life quality index(DLQI) score,the expression of neutrophil elastase(NE),Trappin-2 and placental calcadin(P-cad)in serum and tissue fluid of skin lesion were observed before and after treatment. Recurrence followed up for 6 months in total effective patients and the occurrence of ADR were observed. RESULTS:Two patients of observation group withdrew from the study,and 47 patients completed the study;three patients of control group withdrew from the study,and 45 patients completed the study. Total response rate of observation group was 97.87%,which was significantly higher than 84.44% of control group,with statistical significance(P<0.05). After treatment,PASI,VAS and DLQI scores,the content of NE,Trappin-2 and P-cad in serum and tissue liquid of skin lesion were significantly lower than before treatment;the observation group was significantly lower than the control group. After 6-month follow-up,recurrence rate of total effective patients in observation group(28.26%)was significantly lower than control group(47.36%),and average recurrence time of observation group [(17.91 ± 3.10)weeks]was longer than that of control group [(9.80± 3.41)weeks],with statistical significance(P<0.05). There was no statistical significance in the total incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:Zidan yinxie granules combined with Acitretion capsule and compound flumetasone ointment show clinical efficacy in the treatment of psoriasis vulgaris by lowering the expression of NE, Trappin-2 and P-cad in serum and tissue liquid of skin lesion with good safety.

Objective To investigate the preventive and inhibitory effects of tumor cell lysate(TCL) combined with IL-2 on melanoma and the potential immune mechanism. Methods The B16F10 melanoma TCL cell were prepared using an ultrasonic disruptor. Twenty-four C57BL/6 mice were randomly divided into four groups which were immunized with PBS, IL-2, TCL and TCL+IL-2 for three weeks, and contra lateral tumors were implanted in the fourth week. We observed onset time of tumor and tumor size, collected peripheral blood continuously and monitored the expression of CD4⁺T and CD8⁺T cell subsets dynamically by flow cytometry. Spleen and tumor tissues of mice were also tested for CD4⁺T and CD8⁺T cell subsets by flow cytometry and immunohistochemistry, respectively. Results The preventive immunization of the TCL+IL-2 group significantly delayed the onset time of tumor (P=0.034); moreover, the tumor volume (P=0.023) and tumor weight (P=0.0015) were also significantly smaller than those in the control group. The expression of CD8⁺T cell subsets in the TCL+IL-2 group and the TCL-only group were significantly higher than that in the control group (P=0.0016, P=0.012). However, the CD4⁺T cell subsets of the TCL+IL-2 group decreased after tumor implantation, compared with the control group (P=0.0089). The expression of CD4⁺T and CD8⁺T cell subsets in spleen and tumor tissues were as same as those in peripheral blood. Conclusion The tumor vaccine of TCL combined with IL-2 could prevent the occurrence of melanoma in mouse and effectively inhibit tumor growth by activating CD8⁺T cells.