Objective To prepare optimization of notoginsenoside R1 chitosan nanoparticles,to provide a theoretical basis for clinical application of the drug.Methods Notoginsenoside R1 chitosan nanoparticles were prepared,HPLC method was used to detect the content of notoginsenoside R1 chitosan nanoparticles,preparation technology of nanoparticles were optimized by orthogonal experiment,and the optimized preparation technology of nanoparticles was verified.Results HPLC standard curve equation was A =911.49C -1803.4(r =0.999 9),linear range was from 25 to 900 g/mL.The intra day precision were 1.520%,0.884% and 0.969%(n =6),and the inter day precision were 1.591%,1.447% and 1.269%(n =6).The recovery rates of low,medium and high concentra-tions were (98.11 ±1.16)%,(101.27 ±0.59)% and (100.97 ±0.82)%.4 factors of orthogonal experiment:the concentration of chitosan,the mass ratio of drug and carrier,temperature and rotational speed,and 3 levels of each factor were selected.The average particle size,encapsulation efficiency and drug loading were selected as control indexes.The test results were determined by the method of comprehensive weighted scoring.The orthogonal design was designed according to L9(34)orthogonal design.The optimization process was 2% of chitosan concentration,20% of the weight ratio of drug and carrier,35 ℃ of temperature,600 r/min of rotational speed.According to the optimized process,the average particle size was (123.40 ±7.68)nm,the encapsulation efficiency was (58.41 ±1.59)%,and the drug loading amount was (10.46 ±0.53)%.Conclusion The optimized preparation process of notoginsenoside R1 chitosan nanoparticles is simple and easy to operate,the entrapment efficiency and drug loading amount were high. As a new dosage form,it has a good clinical application prospect.

Objective To screen and optimize the preparation process of Radix Ginseng Capsules.This paper compares the reflux extraction and effects of infiltration percolation extraction to the renshenshouwu capsules.Methods Based on the Orthogonal test,the content of ginsenosides Rg1 and Stilbene glycoside were measured by HPLC.The transferring rates of ginsenosides Rg1 and Stilbene glycoside (1:1 )were regarded as the index to evaluate extracting conditions ,such as the ethanol concentration (%),consumption of alcohol (times),extraction time and extraction times (times). Results The optimum extract conditions of ginsenosides Rg1 and Stilbene glycoside are:with 8 times volume of 50%concentration ethanol as the extract liquid,to refluxing extract 1 times,1.5 h every time.Conclusion The preparation method is simple,practical,high stability and provides the basis for industrial production.

The esophageal cancer mouse models have three categories:chemical carcinogenesis model,xenograft tumor model and genetic engineering model.Chemical carcinogenesis model can simulate well the pathological processes of human esophageal cancer and is prepared simply.Xenograft tumor model is currently the most commonly used esophageal cancer animal model,which has advantages of rapid tumor formation,higher tumor formation rate and lower cost.Genetic engineering model involves mainly p53,Cyclin D1,Brca1 and p120ctn,and it helps to understand the biological characteristics of tumors,but it is expensive and the preparation time is long.

Objective:To compare the detective effect between color Doppler twinkling artifact and B-mode ultrasonography for kidney stones in order to develop an ultrasonic model that can more easily and more accurately detect kidney stone.Methods: 9 patients with kidney stone who were confirmed by CT scan in recently were selected to accepted two methods (color Doppler twinkling artifact and B-mode ultrasonography) to detect kidney stone. The video fragments of upper pole, interpolar and lower pole were created by three radiologists and the results were diagnosed by them.Results: In dual kidneys of the nine patients, 32 stones with average size of (8.9±7.5) mm were found. The sensitivity, specificity, positive predictive value and negative predictive value of the B-mode ultrasonography were 71%, 48%, 52% and 68%, respectively, while those of color Doppler twinkling artifact were 56%, 74%, 62% and 68%, respectively.Conclusions: In the detection of kidney stones, B-mode ultrasonography is more sensitive when is used alone, but color Doppler twinkling artifact has more high specificity. The color Doppler twinkling artifact should be applied in more clinical practices, and a new commercial ultrasonography mode that used basic acoustic difference and is special for detection of kidney stone should be researched and developed so as to improve the detection of kidney stone.

Radiotherapy is one of the most important treatment method for esophageal cancer. Cene expression profile of cancer cells may be one of key factors in determining the effectiveness of radiation therapy To provide guidance for individual therapy, research advances in gene expression involved in radiosensitivity of esophageal cancer are discussed from three aspects: radiation damage repair, cell cycle distribution and signal traneduetion.

Objective To investigate the effects of monocyte CD141 and platelet CD36 on peripheral vascular endothelial function (VEF) in patients with hypothyroidism (PWH). Methods The number of mono-cyte CD141and platelet CD36 from 52 patients wtih PWH and 40 healthy persons (control group) was calculated by flow cytometry. VEF was detected in terms of brachial artery flow-mediated dilatation (FMD) using Doppler ultrasonography. The differences of expressions of monocyte CD141 and platelet CD36 were compared between the two groups, so was FMD. The association of monocyte CD141 and platelet CD36 with FMD (%) was analyzed by Pearson analysis. Results Expression of monocyte CD141 (%) was significantly higher in the PWH group than in the control group (21.79 ± 9.45 vs. 18.84 ± 9.64, P < 0.001), so was expression of platelet CD36 (%) (34.26 ± 10.23 vs. 28.65 ± 9.53, P < 0.001). FMD% was lower in the PWH group than in the control group (8.65 ± 1.97 vs. 11.25 ± 2.72, P < 0.001). CD141 was negatively correlated with FMD% (r = -0.657, P < 0.01), so was CD36 (r = -0.557, P < 0.01). Conclusions Both monocyte CD141 and platelet CD36 are significantly higher in the PWH group than in the control group. CD141 and platelet CD36 are negatively related with VEF.

Objective To investigate the characteristics of hepatitis C viurs (HCV)infection and its genotypes in Yancheng area . Methods A total of 20 185 cases of subjects receiving healthy examination were collected ,and fasting blood levels of serum anti‐HCV were detected .Clinical data of patients with HCV infection were statistically analysed .HCV genotypes and levels of HCV RNA were detected ,and their clinical prognosis was judged by type‐B ultrasonic .Results The total infection rate of HCV was 1 .22% .The infection rate of male was higher than that of female and the infection rate was increased with the elevation of age .The genotype 1b was accounted for 73 .17% .The results of type‐B ultrasonic shown that all patients infected with genotype 6 and 1b/2a HCV only had liver damage .80 .77% of patients infected with genotype 2a HCV had liver damage ,which was higher than that of patients infected with 16 and 3a+3b genotypes .Conclusion Most of HCV infected patients are male ,and the infection rate might be increased with the elevation of age .The prognosis is in various different genotypes of HCV ,which indicates that the prognosis could be evaluated by genotyping .

Objective To probe into the gene therapy of psoriasis using antisense oligonucleotides to attenuate the expression of K14 gene and protein in keratinocytes and evaluate the inhibitory effects of liposome conjugated antisense oligonucleotides on the proliferation of keratinocytes. Methods The antisense, sense and mismatched oligonucleotides for K14 gene were synthesized and conjugated with lipofectin respectively. Finally they were subsequently transfected into cultured keratinocytes in vitro. The expression of K14 gene was tested by reverse transcription polymerase chain reaction (RT-PCR). The expression of K14 protein was measured by immunohistochemistry. The variation of cell growth cycle was detected by flow cytometry. Results The expression of K14 gene and protein was markedly decreased in keratinocytes treated with K14 antisense oligonucleotides. The cell growth cycle was inhibited effectively by antisense oligonucleotides with lipofection, but not by sense and mismatched oligonucleotides. Conclusions Antisense oligonucleotides conjugated with lipofectin might be a hopeful method to inhibit the proliferation of keratinocytes by inhibiting the expression of K14 mRNA and protein.

Objective To investigate the immunological effects of human keratin 17 (K17) on T lymphocytes from psoriatic patients. Methods Total RNA was extracted from the epidermis of psoriatic lesions, and mRNA was reversely transcribed to cDNA. The K17 gene was amplified by PCR and inserted into pGEX-4T-1 expression vector, and then K17 was expressed and purified. T cells were isolated and purified by gra-dient centrifugation and AET-SRBC (2-aminoethylosothiouronium bromide-treated sheep red blood cells) precipitation. MTT assay and direct cell counting method were used to detect the proliferation of T cells. The concentration of IFN- in culture supernatant was measured by ELISA. Results Compared with the control group, K17 significantly enhanced the proliferation of T cells from psoriatic patients and increased the expression level of IFN- (P

BACKGROUND:Study on antisense drug is still one of hotspots in the current field of biomedicine. Due to high-efficiency and specificity, antisense drug used for gene therapy has been paid more attention by many scholars.OBJECTIVE: To observe the effect of liposome-mediated keratin 14 (K14) antisense oligonucleotide on K14 gene and protein expressions as well as in vitro proliferation activities in human keratinocytes (KC).DESIGN: Single sample observation.SETTING: Department of Dermatology, Xijing Hospital, Fourth Military Medical University of Chinese PLA.MATERIALS: Human KC, K14 oligonucleotide gene fragments (modified with phosphrothioate, and above sequence was synthesized by Shanghai Shenggong Bioengineering Company). Reverse transcriptase and TaqDNA polymerase were purchased from Invitrogen Company, K14 monoclonal antibody was purchased from Antibody Company, and SABC kit was purchased from Boster Company. EPICS-PRO-FILE Ⅱ flow cytometer was purchased from Coulter Company (USA).METHODS: Human epidermal KCs were primarily cultured, and their 3rd to 10th generations were used for the experiment.Artificially synthesized sense and antisense as well as mismatched K14 oligonucleotide gene fragments were introduced into KCs by means of liposome. Blank control group were set. The effects of antisense oligonucleotide on the cell cycle,K14 gene and protein expressions of KCs were detected by flow cytometer, reverse transcription polymerase chain reaction and SABC methods.MAIN OUTCOME MEASURES: Effect of oligonucleotide transfecting human KCs on the proliferation of KCs and K14 expression.RESULTS: [1]The electrophoresis of reverse transcription polymerase chain reaction products: Specific K14 gene band appeared in each group, and K14 gene expression in the antisense group was significantly lower in the sense group,missense group and blank control group. K14/β-actin value was similar among sense group, missense group and blank control group (P ＞ 0.05), But K14/β-actin value was significantly lower in the antisense group than in the above-mentioned 3 groups (F =47.554, P ＜ 0.01). ②K14 protein expression detected by immunohistochemical method:K14 was expressed in all the cultured KCs at different levels, and was obviously reduced after antisense oligonucleotide being added. 20 μmol/L antisense oligonucleotide could markedly inhibit K14 expression; K14 expression did not change in the control group. ③ DNA level change detected by flow cytometer: After being treated by K14 antisense oligonucleotide for 48 hours, human epidermal KCs were significantly increased at G1 stage (74.6%), and were markedly decreased at S stage (19.4%). Such changes were not found in the antisense group, missense group and blank control group.CONCLUSION: Antisense oligonucleotide can specifically inhibit K14 synthesis, and thereby, inhibit the proliferation of human KCs.