Objective To develop a scale for evaluating the curative effect of traditional Chinese medicine (TCM) for liver disease under the guidance of the theory of TCM, and to improve the TCM efficacy evaluation standard system for liver disease. Methods According to the TCM theory, the concept model of the liver disease-specific scale was built in combination with the international connotation of QOL and patient-reported outcomes ( PRO). A primary clinical scale was established after item pool screening, field test, culture adaptation and clinical pre-survey by the research staff. Finally, 200 questionnaires were distributed and 198 questionnaires were reclaimed. Statistical analysis was conducted to choose the items. Results All the selected items were analyzed by the different statistical methods, including significance grading, discrete tendency ( coefficient of variation ) method, stepwise regression analysis, discriminatory analysis, Cronbach’s al pha, and principal component analysis and factor analysis. A self-reported subjective scale and a perceived objective scale were developed, including 17 self-reported subjective items and 13 objective items. Conclusion According to the TCM theory and the international connotation of QOL, the scale of assessing curative effect of TCM for liver disease is practical and repeatable. It will be worthy of further evaluation and promotion to clinical application.

Objective To clone and express the polymorphic membrane protein I(PmpI)gene of Chlamydia trachomatis(Ct), and to assess the immunogenicity and biological characteristics of PmpI. Methods A bioinformatic software was used to analyze the sequence of the PmpI gene of Ct, and to predict B cell epitopes in PmpI. With Ct serovar D DNA as the template, PCR was performed to amplify the N?terminal region(from position 90 to 1464)of the PmpI gene, which was cloned into a prokaryotic expression vector pET28a to express the recombinant protein PmpI. A Ni?ion affinity chromatography column was used to purify the recombinant protein, which was used to immunize New Zealand rabbits for preparation of polyclonal antibodies. Western blot analysis was conducted to evaluate the immunogenicity of this protein. Results A comprehensive analysis was carried out on the secondary structure, flexible regions, hydrophilicity plot, antigenic index and surface probability plot of the protein, which suggested that PmpI had 8 dominant B?cell epitopes. The product of PCR targeting the PmpI gene of Ct serovar D showed a total length of 1 375 bp. The recombinant prokaryotic expression vector pET28a?PmpI was successfully constructed. A recombi?nant protein with a relative molecular mass of approximately 50 000 was successfully expressed after isopropylβ?d?1?thiogalactopyranoside (IPTG) induction, and purified by affinity chromatography. Polyclonal antibodies against the recombinant protein were successfully prepared. Conclusion The N?PmpI protein of Ct serovar D is cloned and expressed successfully, laying a foundation for further studies on its biological functions.

[Objective] To investigate the variation of Q promoter (Qp) in nasopharyngeal carcinoma (NPC) cells, and to compare the existing two mutant sites [62 225 site(g→a)and 62 422 site (g→c) ] Qp in NPC cells with the Qp in B95.8 cell line in the functional and biological difference. [Methods] The Qp sequence was amplified in the samples from 29 cases of paraffin-embedded tissues of NPC suffers and 14 cases of peripheral blood of healthy adults by polymerase chain reaction (PCR) method (totally 43 cases). The point mutations on specified sites were analyzed and statistically compared from sequencing results. The sequences of variant and prototype Qp were amplified by PCR and cloned into luciferase reporter vector (pGL3-basic), then transfected into HaCat cells respectively. The transcriptional activity was compared between variant and prototype Qp using luciferase reporter system. The DNA binding affinity of mutant and prototype Qp to Sp1 was compared through chromatin immunoprecipitation (CHIP) method since mutation of nt 62 225 located in a Spl binding site. [Results] The mutation rate of Qp was significantly higher in NPC compared with healthy controls (P=0.039 5, <0.05), which suggested the variant Qp was closely associated with NPC. The transcription of the luciferase gene promoted by variant Qp was significant more than that of prototype Qp in transient transfection assay (2.5:1, P<0.05). The binding affinity of variant Qp to Sp1 was about 1.52 times higher than that of prototype Qp as determined by quantitative ChIP assay. [Conclusions] The transcriptional activity was enhanced in variant Qp in NPC cells compared with prototype, which possibly through the higher binding affinity to Sp1. We suggest that the mutated Qp may play an important role during the EBV infection and transformation of nasopharyngeal epithelium.

Objective To explore the prognostic value of APACHE Ⅱscore in patients with Severe acute organophosphorus poi-soning .Methods 42 patients with Severe acute organophosphorus poisoning ,in which 34 cases survived ,8 cases dead ,were select-ed .The APACHE Ⅱscores of patients in first 24 h of admission were collected ,and receiver operating characteristic curves (ROC curve) were drawn .Results APACHE Ⅱ score of the 42 patients with Severe acute organophosphorus poisoning was 18 ~30 (20 .11 ± 6 .32) ,in which the survival group was(16 .10 ± 3 .12) ,the dead group was(28 .01 ± 4 .46) (P<0 .01) .With the increase of APACHE Ⅱ score ,the fatality rate gradually increased .The total area under the ROC curves of APACHE Ⅱ score for death judgment was 0 .922 ,APACHE Ⅱ score of 21 .2 was the best diagnostic point ,the sensitivity was 95% ,and specificity was 89% . Conclusion The APACHE Ⅱscore could predict severity of patients with Severe acute organophosphorus poisoning ,and APACHEⅡscore ≥21 .2 could be used as the prognosis for death of the patients .

Two sample pretreatment methods of pesticide residues in Panax notoginseng of Chinese traditional medicine were developed. For Method I, the residues were extracted from homogenized tissue with n-hexane-dichloromethane (6:4) by means of ultrasonication, the crude extract was purified by an Envi-carb/NH2 solid-phase extraction (SPE) column. For Method II, matrix solid-phase dispersion (MSPD) technique was used for extracting and cleaning up. The eluates were concentrated by rotary evaporation, and then were redissolved in dichloromethane prior to GC-MS determination. The determination was performed in selected ion monitoring (SIM) mode with the external calibration for quantitative analysis. Under the optimal conditions, the results indicated that the methods are easier and faster, the recoveries of method I for the spiked standards at concentration of 0.01, 0.5, and 2.0 mg x kg(-1) were 81.90%-102.10% with the relative standard deviations (RSDs) of 3.60%-7.10%. The recoveries of method II were 96.26%-104.20% with the RSDs of 3.52%-7.94%. The detection limits (S/N) for residues of pesticides were in the range of 0.48-1.34 ng x g(-1). The results indicated that these multiresidue analysis methods can meet the requirements for determination of residue pesticides and can be appropriate for trace analysis of residue pesticides in Panax notoginseng.

Objective To observe the family cohesion, cancer pain sensation and belief-perception of patients with primary liver cancer and to analyze the correlations among them. Methods From December 2014 to December 2017, we selected 129 inpatients with primary liver cancer of the First Bethune Hospital of Jilin University as subjects by convenience sampling. The family cohesion, cancer pain sensation and belief-perception of patients were evaluated with the family adaptability and cohesion evaluation scale, second edition (FACESⅡ), Pain Intensity Numerical Rating Scale (PINRS) and Pain Beliefs and Perceptions Inventory (PBPI). Besides, Pearson correlation analysis was used to analyze the correlations among them. Results The scores of family cohesion and pain sensation of patients with primary liver cancer were (56.00±5.12) and (5.53±1.18) respectively. In pain belief and perception, the highest score was (1.35±0.05) in the dimension of "feeling pain is very mysterious". Pearson correlation analysis showed that primary liver cancer patients' family cohesion had negative correlations with cancer pain as well as the items of pain belief and perception (r=-0.426, -0.523,-0.434, -0.451, -0.420;P＜0.05). Conclusions The family cohesion of primary liver cancer patients is in a low level affecting the cancer pain sensation and belief-perception of patients, which suggests nurses attach importance to patients' family cohesion, encourage family members to pay attention to patients and increase communication so as to improve treatment effects of patients.

Polyethylenimine (PEI) is one of the most characterized non-viral vectors. It can condense DNA in a good manner and achieve high transfection efficiency. Minicircle DNA (mc-DNA) is a novel kind of supercoiled DNA which is devoid of bacterial backbone. mc-DNA is superior to conventional DNA for its higher transfection efficiency and longer time-span. In this study, we combined PEI and mc-DNA in gene delivery system. We investigated the physicochemical and biochemical effects of this non-viral system and further explore its potential in tumor gene therapy. mc-DNA was obtained by recombination of parental plasmid in the presence of L-arabinose, and complexed with PEI. The results of transmission electron microscopy and scanning electron microscopy showed that the particles were spherical and homogeneous. Through gel retardation assay and MTT assay, we found that there were no obvious differences in binding capability of PEI to mc-DNA and plasmid DNA, as well as in cytotoxicity. The results of dynamic light scattering showed that the size of PEI/mc-DNA was about 68 nm, a slight larger than that of PEI/plasmid DNA. Furthermore, the tumor cells transfected with mc-GFP showed higher GFP expression level than that of conventional plasmid. The same results were achieved in the cells treated with tumor-suppressor gene pten, assayed by RT-PCR and Western blot. It indicates that the system of PEI/minicircle DNA is promising in gene transfer.
DNA, Circular ; genetics ; Gene Transfer Techniques ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Nanoparticles ; chemistry ; PTEN Phosphohydrolase ; genetics ; Polyethyleneimine ; metabolism ; Transfection

Radiotherapy is a treatment for cancer with undesired by-effects. In order to develop a new radiation protective agent that could reduce the by-effects, we tried to express and purify human cryptochrome 1 (hCRY1). The coding sequence of hCRY1 was inserted into prokaryotic expression plasmid pET28a(+), and this protein was purified from Escherichia coli BL21(DE3) after IPTG induction, ultrasonication, inclusion body dissolution, gradient dialysis, nickel column purification and ultrafiltration. The yield of hCRY1 in 1 L E. coli culture (LB medium) was about 10-15 mg. The radiation protective efficiency of hCRY1 was monitored by detecting X-ray-induced H2A.X foci in HaCaT cells. The results of immunofluorescence show that hCRY1 significantly reduces X-ray stimulated DNA damage response. The apoptosis of HaCaT cell was also detected, and the repression of H2A.X foci formation was not due to hCRY1's cytotoxity. All these data suggest a potential application of recombinant hCRY1 as a protective agent for radiotherapy.
Cryptochromes ; biosynthesis ; Escherichia coli ; Humans ; Plasmids ; Radiation-Protective Agents ; Recombinant Proteins ; biosynthesis

RNA silencing is a conserved eukaryotic pathway involved in the suppression of gene expression via sequence-specific interactions that are mediated by 21-23 nt RNA molecules. During infection, RNAi can act as an innate immune system to defend against viruses. As a counter-defensive strategy, silencing suppressors are encoded by viruses to inhibit various stages of the silencing process. These suppressors are diverse in sequence and structure and act via different mechanisms. In this review, we discuss whether RNAi is a defensive strategy in mammalian host cells and whether silencing suppressors can be encoded by mammalian viruses. We also review the modes of action proposed for some silencing suppressors.
Animals ; Gene Expression Regulation, Viral ; Gene Silencing ; Host-Pathogen Interactions ; Humans ; Mammals ; virology ; MicroRNAs ; genetics ; metabolism ; Plant Viruses ; physiology ; Plants ; virology ; RNA, Small Interfering ; genetics ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism ; Viruses ; growth & development

Objective To explore the clinical nursing effect of nursing intervention in elderly patients with hypertension.Methods A total of 100 elderly hypertensive patients were randomly divided into two groups.The patients in the control group were given routine nursing,and patients in the experimental group were treated with systematic nursing.Compliance,blood pressure,blood glucose,blood lipid level,social support,hypertension knowledge score and satisfaction were compared before and after treatment.Results The compliance after intervention in the experimental group was significantly higher than that in the control group,SBP,DBP and 2 h PPG levels were significantly lower than that of the control group,and the total score of social support after intervention was significantly higher than that of the control group.K score,A score and KAB scores of experimental group were significantly higher than that of control group,and B score was significantly lower than that of the control group.The overall satisfaction of experimental group was significantly better than that of the control group,and there was significant difference (P ＜ 0.01).Conclusion Nursing effect of nursing intervention in elderly patients with hypertension is significant,and it has draw great significance.