Objective To optimize the extracting process of the mixture of Feining.Methods Orthogonal design was used to optimize extracting process.The content of glycyrrhizicac,Ephedrine and Pseudoephedrin,and the extractive rate of water and n-butyl alcohol were indexes.Results The optimum water-extracting factors were 8 times of water,refluxing and extracting for 1 h,for 2 times.Conclusion The optimal extraction process is precise and repeatable,which can be used in industrial production.

Objective To optimize the extraction process of Xuanfei Zhike Mixture. Methods The preparation process was investigated by L9(34) orthogonal design with content of ephedrine hydrochloride, the extraction rate of n-butanol and total extraction as indexes. The main factors influencing the extraction efficiency such as the amount of added water, the time of extraction and times of boiling were investigated. Results The optimum technical condition was as follows:8 times amount of water, extract 2 times and 60 min for each time. Conclusion The result is reliable. It can be used for mass production.

Objective To develop an efficient method for detecting the short tandem repeat(STR) of fetal DNA in maternal plasma by miniSTR technique.Methods A total of 9 blood samples form pregnant women from 11 to 27 weeks of gestation were collected.Each isolated total plasma DNA was amplified in single multiplex using the ABI MiniFilerTM kit,which could simultaneously genotype the 9 miniSTR loci,including D13S317,D7S820,D2S1338,D21S11,D16S539,D18S51,CSFIPO,FGA and Amelogenin,and the PCR products were detected by using ABI PrismTM 3100 DNA Sequencer.The allelic designation of each STR locus was accomplished using the GeneMapper ID 3.2 software.Results Father-origin fetal STR allele was detected in all the 9 plasma DNA samples.An average of 3.1 fetal STR alleles of the 8 autosomal STR loci was observed in each of the 9 plasma DNA samples.As for the Amelogenin locus,Amelogenin Y allele was detected in 5 plasma DNA samples from pregnancies with male fetus,and allelic peak height values were all over 50 RFU,according to ABI Mini FilerTM PCR conditions,and the ratio of Amelogenin Y allele peak height value to Amelogenin X allele peak height value was 8.45%.However,no Amelogenin Y allele was detected in other 4 plasma DNA samples from pregnant women with female fetus.Conclusion The miniSTR technique is suitable for STR genotyping using fetal DNA in maternal plasma,and it suggests a broad application in noninvasive molecular prenatal diagnosis.

Objective To explore the role of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in hyperoxia-induced lung injury in preterm rats. Methods At the 2 nd postnatal day Sprague-Dawley preterm rats were randomly assigned to air group and hyperoxia group (exposed to about 85% of O 2). At 3,7,14 and 21 days after exposure, six rats of each group were used to assess lung histologic changes and expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 in lungs by immunohistochemistry. At 3,7 and 14 days after exposure, gelatinase activity in bronchoalveolar lavage fluid (BALF) of another six rats in each group by gelatin zymography was examed. Results Except 3 d after exposure, hyperoxia group showed lung injury characterized by subacute alveolitis and inhibition of lung development. Expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 in hyperoxia group was stronger than that in air group at every time (P

OBJECTIVE:To establish the quality standard of Xiaofeng granules.METHODS:Rehmannia glutinosa,Schizonepetae tenuifolia,Radix et Rhizoma Glycyrrhizae,Rehmanniae glutinosa in the formulation were identified by TLC,and the content of decloxizine hydrochloride in Xiaofeng granules was determined by HPLC.RESULTS:The TLC characteristics were distinctive.At the corresponding position,the sample and its reference substance showed identical color of TLC spots,and there was no interference from negative control.The linear range of decloxizine hydrochloride was 2.45～78.4 ?g?mL-1(r=0.999 9)and its average recovery was 99.11%(RSD=1.30%,n=6).CONCLUSION:The established quality standard can be used for the quality control of Xiaofeng granules.

Background and purpose:Cervix cancer is the second most common cancer in gynecologic malignant tumors. Chemotherapy has been used more and more lately, and concurrent chemo-radiotherapy are receiving increasing attention. In this study, we investigated the clinical effect of concurrent PCF chemotherapy plus radiotherapy for moderate and advanced cervical cancer. Methods:One hundred and ninty-six patients with moderate and advanced cervical cancer were randomly divided into two groups: only radiotherapy group and chemo-radiotherapy group (98 cases for each group). Two groups underwent the same fractionation irradiation, the chemo-radiotherapy group was given PCF regimen that consisted of (DDP) at the dose of 20 mg daily intravenously for 1-5 days, (5-FU) at the dose of 500 mg daily for 1,3,5 days and (CTX) at the dose of 400 mg daily for 2,4 day for 2-4 cycles. Both groups received radiotherapy with 30 Gy first and were irradiated by the combination of external radiotherapy plus intracavitary irradiation.Results:The immediate response rates of the two groups were 100 %, there was no statistical difference. The 5 year survival rate was 76.5 % in chemo-radiotherapy group and 50 % in only radiotherapy group (P

Objective To establish a HPLC method for the determination of amygdalin in Semen Pruni. Methods The analytical column was Supelco ODS Cis column {4.6 mm ? 250 mm, 5 ?m), with methanol-water (20 : 80) as mobile phase. The flow rate was 1.0 mL/min, and the detection wavelength was 210 nm. Results The linear range for amygdalin was 0.06613～4.232 ?g (r=1.0000). The average recovery was 95.91% (RSD=2.29%). Conclusion The method is accurate and reliable, and can be used in the determination of amygdalin in Semen Pruni.

Objective:To explore the expressions of miRNA-17-92 (miR-17-92) cluster and mitofusin 2 (MFN2) protein in endometrial cancer (EC) and their clinical significances.Methods:A total of 72 EC tissues, 36 endometrial lesions of patients with endometrial atypical hyperplasia, and 22 normal endometrial tissues from total hysterectomy for grade Ⅲ cervical intraepithelial neoplasia in the Second Affiliated Hospital of Shandong First Medical University from January 2008 to December 2014 were collected; at the same time, all patients' paraffin-embedded tissues were collected. Real-time quantitative polymerase chain reaction was used to detect the expression level of miR-17-92 in each tissue. Immunohistochemical SP method was used to detect the localization and expression level of MFN2 protein in each paraffin-embedded tissue. The correlation of miR-17-92 and MFN2 protein with clinicopathological features of EC patients was analyzed. Kaplan-Meier method was used to draw survival curve of patients with different miR-17-92 and MFN2 levels, and log-rank test was made; Cox proportional hazard regression model was used for multivariate survival analysis.Results:The relative expression of miR-17-92 in EC, atypical hyperplasia and normal endometrial tissues were 1.49±0.46, 1.01±0.30 and 0.69±0.20, respectively. The expression of miR-17-92 in EC tissues was higher than that in the other endometrial tissues, and the differences were statistically significant (both P < 0.01). The high-expression rates of MFN2 protein in EC, atypical hyperplasia and normal endometrial tissues were 20.8% (15/72), 39.4% (13/33) and 85.0% (17/20); the high-expression rate of MFN2 protein in EC tissue was lower than that in the other endometrial tissues, and the differences were statistically significant (both P < 0.012 5). In EC patients, the relative expression of miR-17-92 in patients with histological type Ⅱ was higher than that in patients with histological type Ⅰ ( P < 0.05); the relative expression of miR-17-92 in patients with myometrial invasion depth ≥1/2 were higher than that in patients with myometrial invasion depth <1/2 ( P < 0.05). The high-expression rate of MFN2 protein in patients with histological type Ⅰ was higher than that in patients with histological type Ⅱ ( P < 0.05); the high-expression rate of MFN2 protein in patients with International Federation of Gynecology and Obstetrics (FIGO) grade Ⅰ was higher than that in patients with FIGO grade Ⅱ and Ⅲ ( P < 0.05). When the EC patients were grouped according to the median relative expression of miR-17-92 (1.421), Kaplan-Meier survival analysis showed that the median overall survival (OS) time of the miR-17-92 low-expression group (36 cases) was not reached, and the high-expression group (36 cases) was 36 months (95% CI 32-42 months), and the difference in OS between the two groups was statistically significant ( P = 0.049); the median OS time of the MFN2 high-expression group (15 cases) was not reached, and the low-expression group (57 cases) was 38 months (95% CI 33-41 months), and the difference in OS between the two groups was statistically significant ( P = 0.046). Multivariate Cox regression analysis showed that the expression levels of miR-17-92 and MFN2 were independent influencing factors for the survival of EC patients ( HR = 3.10, 95% CI 1.36-7.07, P = 0.007; HR = 0.30, 95% CI 0.09-0.99, P = 0.048). Conclusion:The high-expression of miR-17-92 and low-expression of MFN2 protein in EC tissues may be involved in the occurrence and development of EC, and they can be used as indicators for judging the prognosis of EC patients.

BACKGROUND: The judgment of the engraftment of hematopoietic stem cells after transplantation mainly depends on various genetic labeling in vivo, which are different in sensitivity and effectiveness, thus a method with powerful differential ability, high sensitivity and not restricted by sex is to be established.OBJECTIVE: To observe the DNA genetic loci of short tandem repeat in the blood samples of both donors and recipients before allo-hematopoietic stem cell transplantation and those of recipients at different time points after transplantation.DESIGN: An observation measurement.SETTING: Laboratory of Immunogenetics, Shenzhen Institute of Transfusion Medicine, Shenzhen Blood Center.PARTICIPANTS: Blood samples of 18 pairs of donors and recipients, who were successfully matched and accepted hematopoietic stem cell transplantation, were selected from the Laboratory of Immunogenetics, Shenzhen Institute of Transfusion Medicine, Shenzhen Blood Center from February 2004 to December 2005. Among the 18 patients, there were 10 males and 8 females, with a mean age of 35 years old, including 6 cases of them were donated by relatives with blood relationship, and 12 cases by volunteers without blood relationship. Informed consents were obtained from all the participants.METHODS: The blood samples of both donors and recipients before transplantation and the blood samples of recipients after transplantation were collected, and the fluorescence labeling short tandem repeat technique was used to detect the 15 loci for short tandem repeat and Amelogenin sex locus, so that the differential loci between the donor and recipient could be screened. The engraftment and dynamic changes of the short tandem repeat genes of the donors in the recipients after transplantation were observed, the times for the earliest occurrences of short tandem repeat genes of the donors and the complete chimerism were recorded.MAIN OUTCOME MEASURES: ① Differential genes between the donors and recipients before transplantation;②Times for the earliest occurrences of short tandem repeat genes of the donors and the complete chimerism.RESULTS: All the 18 pairs of donors and recipients were involved in the final analysis of results. Satisfactory results of the typing at the 15 loci for short tandem repeat and 1 sex locus in the 18 pairs of samples of both donors and recipients before transplantation and the sample of the recipients after transplantation respectively. Averagely 12.4 (8-15) differential loci for short tandem repeat could be distinguished between the donors and recipients. ②After transplantation, short tandem repeat genes could be detected the earliest at 8 (5-14) days averagely, It took 14 (9-23) days averagely for short tandem repeat loci to convert from recipient type completely into donor type, and the engraftment converted from the recipient chimerism types completely into the donor types.CONCLUSION: The fluorescence labeling compound amplification of short tandem repeat technique can precisely measure the number of PCR products, describe the engraftment of hematopoietic stem cells and the whole process of development. It can also provide accurate and timely information for the early judgement of engraftment, predicting failure of transplantation and controlling recurrence.

Objective To study the molecular genetic background of Bel subtype at ABO blood group.Methods Three samples and fifteen samples were diagnosed as Bel subgroup and normal control samples by serological test,respectively.The extracted DNA was genotyped by sequence specific primer- polymerase chain reaction foilowed by sequencing for Exon6 and exon7 at ABO locus and clones were sequenced.Results A novel Bel variant allele(GenBank EF117687) was identified in a Bel individual.The Bel allele was different from the regular B101 allele by single 952G>A missense mutation in exon7.resulting in an amino acid subsfitution of Val for Met at 318 locus.No mutations were detected in the fifteen control samples and the other two Bel allele samples.Conclusions The mutation position was fimt found to lie on coding region of ABO gene behind nucleotide 930.The mutation of G952A in the al,3 galactosyhransferase gene may be one of the molecular genetic basis of Bel ohenotype.