Objective:To Amplify,purify and identify the cDNA library of human K562 cell in E.coli DH5? and to transform it into yeast cell for subsequent target protein screening.Methods:The cDNA library of human K562 cell was amplified.Then the library plasmids were extracted and transformed into Y187 yeast cell.The results of transformation were verified by PCR and restriction enzyme analysis.Results:The cDNA library of human K562 cell was amplified successfully and the diversities were verified.The cDNA library of human K562 cell was also successfully transformed into Y187 yeast cell.Conclusion:Successfully amplification,purification and identification of the cDNA library of human K562 cell lay the foundation for the subsequent protein screening.

Objective To investigate the clinical features of hypotensive maintenance dialysis patients after total parathyroidectomy with autotransplantation (TPTX + AT).Methods The consecutive patients who received TPTX + AT were enrolled between January 1,2010 and May 1,2015 in the nephrology department at the central hospital of Jiangmen.According to level of the blood pressure after TPTX + AT,the patients can be divided into hypotensive-group and non-hypotensive-group.The clinical and laboratory examinations of them were compared.Results The probability of calcification of aortic valve (4.76％ vs 30％,P =0.045),bicuspid valve (9.52％ vs 40％,P =0.032) and pulmonary hypertension (14.29％ vs 45％,P =0.043) was lower in the hypotensive-group with statistical significance.Serum parathyroid hormone [(9.31 ± 5.41) pg/ml vs (60.12 ± 96.95) pg/ml,P =0.021] and angiotensinⅡ (Ang Ⅱ) [(34.26 ± 15.73) pg/ml vs (63.78 ±23.55) pg/ml,P =0.000] were lower in the hypotensive-group with statistical significance.Serum intact parathyroid hormone (iPTH) (P =0.036) and AngⅡ (P =0.002) were the affecting factors of hypotension after TPTX + AT.Conclusions Hypotension after TPTX + AT is associated with the lower level of serum iPTH and Ang Ⅱ.The probability of calcification of cardiac valve and pulmonary hypertension is lower in the patients with hypotension,indicate that they may have better cardiac prognosis.

Objective To investigate the potentiality of specific miRNA level in blood plasma of ovarian cancer patients as a molecular marker to predict sensitivity and response to platinum-based chemotherapy .Methods Quantitative polymerase chain reac-tion ( Q-PCR) was used to check the changing of miRNA level in blood plasma before and after chemotherapy , and explored the rela-tivity between variation and chemotherapy response and clinic outcome .Results Among preliminary screened 11 miRNAs, only 5 miRNAs were able to be detected in peripheral blood .There existed variations in 3 miRNAs ( miR-22, miR-106a, miR-142).Further detection of the changing of miRNA level in blood plasma before and after therapy , miR-22 was found significantly up-regulated in blood plasma that was underwent platinum-based chemotherapy .Meanwhile, ovarian cancer patients with high miR-22 level were most-ly sensitive to platinum-based therapy.Otherwise, miR-22 was positive relative to the clinic prognosis of patients with ovarian cancer . Conclusions miR-22, miR-106a, and miR-142 might participate in the generation and development of ovarian cancer .miR-22 in pe-ripheral blood was probably identified as a novel molecular marker of prediction of the sensitivity to platinum in ovarian cancer patients .

Objective To observe the therapeutic efficacy and safety of recombinant tissue plasminogen activator (rt-PA) for treatment of patients with acute cerebral infarction and investigate the prognostic factors of thrombolysis. Methods The clinical data of 60 patients with acute cerebral infarction from January 2009 to November 2013 in Department of Emergency, Hangzhou Traditional Chinese Medicine Hospital were retrospectively analyzed;of them, 30 cases received intravenous rt-PA thrombolytic treatment, being in the thrombolytic group, 0.9 mg/kg rt-PA was given to the patient within 4.5 hours after the disease onset, the total dosage could not exceed 90 mg, in which 10%was intravenously injected and the rest 90%was intravenously dripped slowly within 60 minutes. That another 30 cases did not undergo thrombolytic therapy was assigned as the control group, and they took aspirin, etc anti-platelet aggregation routine treatment. Before and after thrombolytic treatment for 1 hour, 24 hours and 14 days, the National Institutes of Health Stroke Scale (NIHSS) score was evaluated in the two groups;before and after thrombolytic therapy for 0.5, 1.0, and 1.5 hours, the patient's scores of Barthel index (BI) were observed in the two groups. In thrombolytic group, the situations of hemorrhage transformation, symptomatic hemorrhage and modified Rankin scale (mRS) score in 3 months of the patients with different ages, complications and NIHSS scores were observed. Results Before treatment, no statistically significant differences were found in the NIHSS score (15.2±3.6 vs. 15.5±3.3) and BI score (45.0±8.8 vs. 44.1±7.6) between the control group and thrombolytic group (both P>0.05);after treatment with the extension of time, the NIHSS score was gradually reduced, reaching the lowest level on the 14th day after thrombolytic treatment, while the BI score was gradually increased, reaching its peak at 1.5 hours after thrombolytic treatment, the changes being more prominent in thrombolytic group (NIHSS score:9.7±2.6 vs. 12.8±4.2, BI score:82.6±7.8 vs. 69.6±9.8, both P<0.05). In thrombolytic group, there were cerebral hemorrhage transformation 2 cases, gum bleeding 3 cases, skin bleeding 1 case, urethral bleeding 1 case, gastrointestinal bleeding and black stool 1 case. In control group, cerebral hemorrhage transformation was seen in 1 case. There was no obvious systemic hemorrhage in the two groups. In the 30 cases in thrombolytic group, the baseline NIHSS score of patients>80 years old was higher than that in cases≤80 years old (15 vs. 12); the age (years: 71.0±4.1 vs. 61.5±2.6), baseline NIHSS score (14 vs. 11) and bleeding conversion rate [37.50% (3/8) vs. 18.18% (4/22)] of cases with atrial fibrillation were higher than those not complicated with atrial fibrillation; the NIHSS score of cases with elevated international normalized ratio (INR) was lower than those without elevated INR (11 vs. 14);The bleeding conversion rate [16.67%(1/6) vs. 29.17%(7/24)] with NIHSS score≤4 were lower than those with NIHSS score>4;the mRS score in 3 months (4 vs. 2), and percentage of 3-month mRS score≤2 [11.1%(1/9) vs. 52.38%(11/21)] in cases with NIHSS score≥20 was higher than that in cases with NIHSS score<20 (all P<0.05). The fatality rate of two groups was 3.33%. Conclusions The intravenous rt-PA thrombolytic treatment can significantly promote the early recovery of neurologic impairment for patients with acute cerebral infarction. The therapy can improve the prognosis and its safety is relatively good. Meanwhile it is similarly effective for cases over 80, with complications such as atrial fibrillation, raised INR, and with different degrees of severity.

Janus kinase 2,myeloproliferative leukemia virus and tet encogene family member2 mutations affect a variety of cytokines signal transduction pathway in BCR/ABL-negative myeloproliferative neoplasms (MPN). In the influence of mutation load,co-mutation and genetic susceptibility,these mutations can induce different MPN phenotypes,and affect the characteristics of patients,the distribution of peripheral blood cells and prognosis. But how these mutations contribute to disease initiation,development,and transformation needs further reseach.

Objective: To reconstruct a substractive complementary deoxyribornucleic acid (cDNA) library of genes sensitive to mechanical stretch in human osteoblast like cells.Methods: Mechanical stretch at 12 cycles per minute was applied to human osteoblast like cells Saos-2 and the deformation of the stretched cells was 12%. Twelve hours after loading, mRNAs were isolated from both stretched and unstretched cells. Substractive cDNA library of the genes sensitive to stretch was constructed with the technique based on polymerase chain reaction (PCR) and substractive hybridization. Primary sequencing of clones in the library was carried out. Results: A substractive cDNA library of genes sensitive to stretch was constructed with a capacity of about 200 clones. According to the results of sequencing, most genes in the library were related to the mechanical stimulation. One novel gene fragment was obtained. Conclusion: The method used in the experiment is effective in cloning genes sensitive to mechanical stretch.

Objective To investigate the role of Th cells in dextran sulfate sodium (DSS) induced murine colitis. Methods The ratios of Th1/Th2 cells intestinal mucosa and splenic monocytes (SMC) of mouse with DSS induced murine colitis were detected by FCM with two color intracellular cytokine staining. Results (1) In lamina propria mucosa cell (LPMC) of murine colonic mucosa from the acute model, Th1 cell was (8.90?1.23)%, significantly higher than that in the control( P 0.05). The ratio of Th1/Th2 was 3.09?1.18, significantly higher than that in the controls( P

A simple and efficient method, based on ultrasound-assisted polymer surfactant-enhanced emulsification microextraction followed by gas chromatography-mass spectrometry( GC-MS) , was developed for the determination of eight aromatic amines ( AAs ) in aqueous sample. The main parameters affecting the performance of the purposed method were optimized, and the optimized conditions were obtained as follows:150 μL of extractive solvent ( dichloroethane ) , polymer surfactant ( sodium alginate ) concentraction of 0. 20 g/L, pH 7. 0, salt addition of 3%, and ultrasound time of 1 min. Under the optimal conditions, the linear ranges of the method were 0. 1-200 μg/L for 3,3’-dichlorobenzidine, 0. 3-200 μg/L for 2,4,5-trimethylaniline, 4-chloro-o-toluidine, 3,3’-dimethyl-4,4’-diaminodiphenylmethane, 3,3’-dimethylbenzidine and 4 , 4’-methylene-bis-( 2-chloroaniline ) , 0 . 5-200 μg/L for 4-aminoazobenzene and 3 , 3’-dimethoxy-benzidine. The correlation coefficients (R2) and the detection limits were 0. 9961-0. 9997 μg/L and 0. 08-0. 3 μg/L, respectively. The intra- and inter-day RSDs were less than 10. 6% and 11. 0%. The purposed method could be applied to the analysis of AAs in water sample collected from tap water and river water. In comparison with the common SA-USAEME, conventional surfactant was replaced with water-soluble polymer surfactant to solve the problems of potential pollution, and the polymer surfactant was natural and insoluble in extractive solvent. Compared with other solid-phase extractions, the proposed method had the advantages such as simple operation and low cost.

Objective To analyze the clinical and radiographic outcomes of staged reduction and fixation in a consecutive series of patients with the old Lisfranc injuries. Methods Fifty patients (16 feet) with Lisfranc injuries were treated with staged reduction. Mean duration between injury and surgery was 4. 8month ( 3 to 8 month) . In first stage an external fixator was applied across the Lisfranc joint and distraction was done at 1 milliliter per day to 2 milliliter per day. In the second staged the ORIF ( open reduction and internal fixation) was doneand we were able to reduce all the fractures and dislocations. Extra-Articular screws and staple fixation were used for fixation. We compared categorical variables using Fisher’ s exact test and continuous variables using paired t-test or Wilcoxon signed-rank test. Results All patients were followed up 1 to 3 years ( mean 2. 2 years) in the clinic. The visual analogue scale score averaged 3. 1 points at the final follow-up, the average AOFAS scores for these patients were 55. 8 points ( range, 43 to 98 points), with a significant increase than before surgery ( P=0. 001). The mean duration between two surgeries was 3. 2 weeks (range 2. 5-4. 5 weeks). Anatomic reduction was obtained in all 15 patients. At the last follow-up, 2 patients had lost reduction. Posttraumatic osteoarthritis was observed in 5 patients, and all of them were scheduled for arthrodesis because of persistent pain. Conclusions The study have displayed that staged reduction and Extra-Articular fixation should be considered for old Lisfranc injuries with a good reduction, the firm stability, low risk of intraoperative fracture. The short-term effectiveness is good, but the long-term effectiveness needs further follow-up.

Objective To discuss the value of ABCD2 score in predicting medium and long-term prognosis in patients with ischemic stroke.Methods 184 patients with ischemic stroke admitted to department of neurology from March 2014 to December 2014 were selected as the research object.According to the ABCD2 score points prior to admission, the patients were divided into 3 groups: 40 cases in the low risk group, 76 in the moderate group and 68 in the high risk group based on whether or to what extent he was able to do self care.The phone call reviews were made for the clinical symptoms of patients discharged from the hospital after six months..According to the modified Rankin scale classification standard, the patients discharged from the hospital after six months were divided into 2 groups: 126 cases in the self-care group and 56 cases in care-needing group.The groups were compared in terms of conditions.Results The self-care rate in the low risk group was 90.00%, the moderate group 77.63% and the high-risk group 45.59%.The care needing rates in the three groups was 5%, 19.74% and 48.53%, respectively.The care needing rate in the low risk group was obviously lower than that in the moderate-risk group and the high risk group (P<0.05).The care needing rate in the moderate risk group was obviously lower than that in the high risk group (P< 0.05).The self-care rates in the low risk group and the moderate group were higher than that in the high risk group (P<0.05).Conclusion The ABCD2 score has a higher predictive value for medium and long-term prognosis in patients with ischemic stroke.