A novel method for quickly cloning genes with multiple DNA fragments--one step cloning technique using isoschizomer-heterotail restriction endonuclease (IHRE) is described. Up to six DNA segments are ligated by using only one restriction endonuclease in this method. Comparing with routine method,it is simple, fast, economical and generates products with higher purity and achievement. Light chain of human enterokinase, DNA multi-epitope vaccine to HSV2 have been designed and successfully constructed via this method.

Objective To study teaching cases of health economics in the context of health reform and development in China.Method Preparing for lessons in groups,information collection,experiences summary,and field research.Result Three productions:a teaching plan,a case collection,and a guideline of case teaching.Discuss It is useful to improving students’ ability to contact theory with practices and enhance teacher’s teaching and researching ability,but it also needs further improvement.

Objective To evaluate the sensitivity, specificity and clinical significance of CK19 mRNA, CEA mRNA and LUNX mRNA for detecting micrometastasis by sampling the peripheral blood and regional lymph nodes of lung cancer patients. Methods Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect LUNX mRNA,CK19 mRNA, CEA mRNA for micrometastasis by sampling the peripheral blood of 48 lung cancer patients and 44 regional lymph nodes of such patients treated by curative resection. Peripheral blood of 30 patients with pulmonary benign lesions and 10 normal healthy volunteers and lymph nodes of 6 patients with benign pulmonary diseases served as control. Results (1) LUNX mRNA, CK19 mRNA, CEA mRNA were expressed in all(35/35) lung cancer tissues. (2) In the peripheral blood from 48 lung cancer patients, 30(62.5%) were positive for LUNX mRNA, 24 (50.0% ) positive for CK19 mRNA and 32(66.7%) positive for CEA mRNA. The positive detection rates of micrometastasis in 44 lymph nodes from lung cancer patients were 36.4% (16 out of 44) for LUNX mRNA, 27.3%(12 out of 44) for CK19 mRNA and 40.9%(18 out of 44) for CEA mRNA. (3) In the 30 blood samples from patients with pulmonary benign diseases, 2 (6.7%) expressed CK19 mRNA, but none expressed LUNX mRNA or CEA mRNA. All the 3 molecular markers were negative in the 10 blood samples from healthy volunteers. In 11 lymph nodes from patients with pulmonary benign lesions, none was positive for any of the three markers.(4)In 44 regional lymph nodes from lung cancer patients,6(13.6%) were positive for metastasis by histopathological examination, with a positive rate significantly lower than that of the RT-PCR ( P

The paper presented the principles and references for identifying services of the primary health care at townships and villages in Beijing, and proposed the screening criteria for primary health care package in rural Beijing. Studies made have identified the screening results for the package applicable to both townships and villages in Beijing, along with analysis for the rationale, applicability and operability of the package. Moreover, it probed into the assurance conditions for offering primary health care as a reference for other regions in the country.

Objective To understand the influence of the disease-based hierarchical medical system on inpatients flow covered by the new rural cooperative medical system ( NRCMS) , and that on the funding diversion and medical costs so incurred. Methods One county was selected from the eastern, central and western regions of China respectively, where the disease-based hierarchical medical system has been in place. Policy documents of the three counties were reviewed to analyze such changes as NRCMS inpatients flow, inpatients subsidy diversion, NRCMS fund surplus rate of the current year and medical costs per hospitalization before and after the system was in place. Results A comparison with 2014 found a 1. 26%drop of the out-of-county inpatients of county W of the western region, a 2. 00% increase of township hospitals inpatients of county D in the middle region, and the same ratio of out-of-county and in-county inpatients in county F of the eastern region in 2015. Compared with 2014, the fund surplus rate of county W increased 10. 46%, and the inpatient subsidy ratio of county D decreased 2. 51% for those in out-of-county medical institutions in 2015. Thanks for the quota payment of specific diseases under global budget in county W, the inpatient medical costs per hospitalization dropped at both county and township medical institutions. Conclusions The disease-based hierarchical medical system could optimize the NRCMS inpatients distribution among various medical institutions, conducive for establishment and operation of such a system.

[Abstrct] Objective To compare the short -term tumor response and adverse events of two chemoradio-therapy regimens for locally advanced non -squamous non-small cell lung cancer .Methods From March 2009 to January 2013 ,we recruited 42 patients with stage ⅢNSCLC who had received three -dimensional intensity -modulated radiotherapy combined with chemotherapy .group A treated with radiotherapy combined pemetrexed plus cisplatin and group B treated with radiotherapy combined docetaxel plus cisplatin .We compared the short -term tumor response and adverse events between the two regimens .Results There were 28 cases in group A and 14 cases in group B .There were no significant differences between the two groups in hematologic toxicities ,such as leukopenia,neutropenia,anemia and thrombocytopenia.In non-hematologic toxicities,radiation pneumonitis and cough were no significances respectively (P<0.05).There were no significant differences in other non -he-matologic toxicities,such as liver dysfunction,renal dysfunction,fever,dyspnea,radiation esophagitis,hypody-namia,weight loss,gastrointestinal reactions and skin reactions .The response rate displayed no differences be-tween two groups .Conclusion This study reveals Pemetrexed plus cisplatin group had less non -hematologic toxicities than Docetaxel plus cisplatin group in locally advanced non -squamous non -small cell lung cancer . But there are no differences in the short -term tumor response between the two regimens .

Radiotherapy not only acts as an important local treatment of cancer, but also plays a key role in regulation of immune function. Radiotherapy regulates anti?tumor immune responses by promoting the generation of neoantigens, regulating the release of cytokine, and enhancing the sensitivity of tumor to cell?mediated immunity. Recently, several studies and clinical practice reported the abscopal effect in some patients undergoing radiotherapy combined with immunotherapy, which showed partial or complete response of metastases outside the irradiation field, suggesting the combination therapy as a promising strategy. However, further studies are needed for the understanding of the mechanism and influencing factors for immunity such as radiation dose and fractionation scheme. This paper reviews the research advances in the mechanism of radiotherapy?immunotherapy interaction and the combination therapy for cancer.

Objective:To investigate the association between p53 codon 72 polymorphism and the prognosis of breast cancer pa-tients receiving chemotherapy and radiotherapy after surgery. Methods:A total of 427 breast cancer patients treated with chemo-radio-therapy after surgery at Beijing Cancer Hospital were selected for this study. Polymerase chain reaction–restriction fragment length polymorphism was adopted to analyze the p53 codon 72 polymorphism. Survival analysis was conducted to compare the disparities of recurrence and survival among the patients with different p53 codon 72 polymorphic variants. Results:The distribution of three geno-types of p53 codon 72 in our cohort is as follows:Pro/Pro 18.3%(78/427), Pro/Arg 44.0%(188/427), and Arg/Arg 37.7%(161/427). No significant difference was observed among the local recurrence-free survival (LRFS), loco-regional recurrence-free survival (LR-RFS), distant disease-free survival (DDFS), and overall survival (OS) among the three genotypes (all P>0.05). Among the 303 estro-gen receptor (ER)-positive patients, OS was significantly better in patients with Arg/Arg genotype than those with Pro/Pro genotype (χ2=6.33, P=0.042). The multivariate analysis showed that the p53 codon 72 polymorphism is an independent factor of prognosis for LRFS, LRRFS, DDFS, and OS of ER-positive patients. For the ER positive patients with Pro/Pro genotype, the local recurrence, local-regional recurrence, distant metastasis, and mortality risks were 5.9 (HR=5.9, 95%CI 1.1-31.1, P=0.036), 3.1 (HR = 3.1, 95% CI 1.1-9.1, P=0.039), 2.8 (HR=2.8, 95% CI 1.3-6.0, P=0.010), and 4.0 (HR=4.0, 95% CI 1.3-12.0, P=0.013) times higher than those with Arg/Arg genotype, respectively. Conclusion:For ER-positive breast cancer patients who underwent surgery and chemo-radiotherapy, the local recurrence, loco-regional recurrence, distant metastasis, and mortality risk with Pro/Pro genotype are significantly higher compared to those with Arg/Arg genotype.

Objective To establish the methods for controlling quality of Yiqi Congming Pills, and to improve the quality standard for the pills. Methods Yiqi Congming Pills were sampled from the products of 3 mainpharmaceutical companies in China market, 2 batched from each company. Radix Puerariae, Radix Astragali, Fructus Viticis, Cortex Phellodendri Chinensis, Radix Codonopsis in Yiqi Congming Pills were observed under microscope for microscopic identification. Radix Astragali, Rhizoma Cimicifugae, Radix Paeoniae Alba, and Cortex Phellodendri Chinensis were identified by thin-layer chromatography (TLC) for qualitative differentiation. The contents of puerarin and paeoniflorin in Yiqi Congming Pills were determined by high performance liquid chromatography(HPLC). Results The microscopic characteristics of Radix Puerariae, Radix Astragali, Fructus Viticis, Cortex Phellodendri Chinensis, Radix Codonopsis in the pills were obvious and were easy for discrimination. The spots in thin-layer chromatography (TLC) chromatograms of Radix Astragali, Rhizoma Cimicifugae, Radix Paeoniae Alba, and Cortex Phellodendri Chinensis were clear without interference of negative reference. Puerarin had a good linearity in the range of 0.00112~3.50676μg, and the average recovery rate was 98.2%(sR=1.4%, N=9 ). Paeoniflorin had a good linearity in the range of 0.00333 ~ 3.328 μg, and the average recovery rate was 98.4%(sR = 1.8%, N = 9). Great differences existed in the contents of puerarin and paeoniflorin of Yiqi Congming Pills from the three pharmaceutical companies. Conclusion The established qualitative and quantitative methods are of accuracy, credibility and repeatability, and are effective for controling the quality of Yiqi Congming Pills.

In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor (LIF) in mice with embryonic implantation dysfunction (EID), 120 Kunming mice post coition were randomized into three groups: normal control group, model group and traditional Chinese medicine group (TCM group) (n=40 in each group). Uterus was collected on the pregnancy day (Pd) 4, 5, 6 after an intravenous injection of Evan's blue. The endometrium was dyed by Evan's blue and the mean points of response were observed on Pd 5. The expression of LIF mRNA and protein was detected by RT-PCR and immunohistochemistry respectively and analyzed statistically by image system. The results showed that the number of implantation sites in model group was remarkably less than in normal control group and TCM group. There was no significant difference between normal control group and TCM group. The expression of LIF mRNA and protein in model group was delayed. Bushenantai recipe could increase the expression of LIF mRNA and protein in endometria of mice with EID. It was suggested that Bushenantai recipe could improve embryo implantation of mice with EID by promoting the endometrial LIF expression and endometrial decidualization.
Blastocyst/cytology ; Embryo Implantation ; Endometrium/*metabolism ; Gene Expression ; Gene Expression Regulation, Developmental ; Leukemia Inhibitory Factor/*biosynthesis ; Leukemia Inhibitory Factor/*genetics ; Medicine, Chinese Traditional ; Models, Biological ; Plant Extracts/pharmacology ; RNA, Messenger/metabolism ; Time Factors