Objective To construct pcDNA3 1(+)GDNF recombinant eukaryotic expression plasmid and to investigate its expression in eukaryotic cells. Methods The coding sequence of GDNF was amplified from rat astrocytes by reverse transcription PCR (RT PCR) and cloned into pcDNA3 1(+) eukaryotic expression vector The recombinant pcDNA3 1(+)GDNF plasmid was then transfected into eukaryotic cells mediated by using Fu Gene 6 method Analysis by restricting enzyme digestion and DNA sequencing were carried out to demonstrate the sequence of the plasmid GDNF protein and its activity were then determined using pcDNA3 1(+)GDNF plasmid transfected eukaryotic cells Results RT PCR product is 640 bp specific segment Analysis by restricting enzyme digestion and DNA sequencing of pcDNA3 1(+)GDNF recombinant showed results from restricting enzyme were 640 bp and 300 bp segments respectively DNA sequencing revealed that GDNF cloning was successful The recombinant plasmid can express active GDNF protein in eukaryotic cells Conclusion The study on the role of both GDNF and gene therapy is significant in the treatment of Parkinson disease

Objective To investigate the relationship between the biological characteristics and the expres-sion of stromlysin (MMP-7) and microvessel density (MVD) in breast cancer tissue and the effect of MMP-7 on MVD of breast cancer. Methods 60 fresh samples were obtained from patients with breast cancer,and then the ex-pression of MMP-7 and vascular endothelial cell CD34 were studied by means of immunohistochemical assay (SP method). The relationship between MMP-7, MVD and age, tumor size, histological type, lymph node metastasis and expression of ER, PR, PCNA, p53 and CerbB2 was studied. The relationship between the biological characteristics and the expression of MMP-7 and MVD in breast cancer were evaluated. Results The positive expression rote of MMP-7 in the d>2 cm group,the group with lymph node metastasis,the group with positive expression of PCNA, p53 and C-erbB-2 (78.1%,74.2%,71.8%,67.6% and 72.2%)were all higher than that in the d≤2 cm group, the group without lymph node metastasis,the group without positive expression of PCNA,p53 and C-erbB-2(32.1%, 41.3%,38.1%,38.5% and 33.3%) (P<0.05). MVD was higher in the positive breast cancer tissue of the group of d>2 cm(34.61±6.97), the group with lymph node metastasis (34.37±7.50), PCNA ( 33.24±8.39), p53 (33.28±8.94), C-erbB-2 (33.55±8.57) than in the negative breast cancer tissue of the d≤2 cm group ( 28.60±9.82), the group without lymph node metastasis (27.48±8.66), PCNA (26.88±7.89), p53 (21.71± 7.59),C-erbB-2(27.42±27.69) (P<0.05). MVD(33.62±7.36)/ high power lens in the MMP-7 positive group was higher than that (27.86±9.45)/high power lens in the MMP-7 of negative group(P<0.05). The posi-tive expression of MMP-7 was correlated with MVD (r=0.380, P<0.05). Conclusion The high expression of MMP-7 and MVD is associated with the development and metastasis of breast cancer. MMP-7 could promote the mi-crovessel development in mammary cancer.

Objective To investigate the mutations of CLCNKB gene in a family with classic Bartter syndrome. Methods Genetic DNA was extracted from peripheral blood leucocytes of family members.The coding exons and intron exon junctions of CLCNKB gene were amplyfied by PCR and sequenced directly.Fifty unrelated healthy subjects were selected to exclude the possibility of polymorphism. Results A heterozygous(missense)mutation(482T>G,L161R)was detected in the exon 4 of patients.The hetemzygous mutation(L161R)was found in the mother,while no mutation was found in the father of this family.L161R had not been reported and was a novel mutation when referring to literatures and human genomic database home and abroad.Conclusion A new CLCNKB gene mutation(L161R)is identified for the first time.

Objective To determine the relationship between normal weight obesity (NOW) and cardiovascular risk factors.MethodsA total of 940 adults who received a health examination in out hospital were recruited in a cross-sectional study,and 407 with a body mass index (BMI) of18.5 to 25 kg/m2were enrolled for further analysis.Body fat percentage ( BF％ ) was measured by bioelectrical impedance analysis (BIA),and the subjects were assigned to the NOW group ( BF％ ≥25％ for male or BF％ ≥35％for female) or the control group ( BF％ ＜ 25％ for male or BF％ ＜ 35％ for female).Cardiovascular risk factors and their detection rates were compared between the two groups by using independent sample t test and x2 test.The correlationbetweenNOW and cardiovascular risk factors was assessedbylogistic regression.Results The prevalence of NOW in men and women were13.1％ and14.9％,respectively.The prevalence of NOW was increased with age ( x2 =6.90,P ＜0.05 ).Systolic blood pressure (SBP),diastolic blood pressure ( DBP ),total cholesterol ( TC ),triglycerides ( TG ),low-density lipoprotein cholesterol ( LDL-C) and serum uric acid (SUA) were significantly increased in the NOW group (t values were 2.97,2.44,2.54,5.09,2.71and 3.91,respectively; all P ＜ 0.05 ) ; whereas high-density lipoprotein cholesterol ( HDL-C) was significantly decreased in the NOW group (t =-3.90,P ＜ 0.05 ).The prevalence of hypertension,hyperglycemia,high triglyceride,low HDL-C,dyslipidmia and hyperuricemia was increased in the NOW group in comparison with the control group ( x2 values were 6.76,5.58,14.50,11.97,10.97 and 8.76,respectively;allP＜ 0.05 ).LogisticregressionshowedNOWincreasedtheriskof hypertension,hyperglycemia,dyslipidmia or hyperuricemia by 2.186,2.120,2.088 or 4.175 times.After adjustment for age and gender,the risk for hyperuricemia was decreased to 3.491,but remained statistically significant higher.Conclusions NOW may be correlated with cardiovascular risk factors,and those with NOW could be at higher risk for cardiovascular diseases.

Cestode Infections ; parasitology ; Humans ; Infant ; Male

Objective To investigate the influencing factors of chronic brucellosis.Methods Nested case control method was used to study newly diagnosed patients (n =600) with brucellosis in a cohort study in 2012.Data of general characterstics,clinical presentation,treatment and prognosis of those patients were collected.These patients were followed up for one year,and the chronic patients as the case group (n =248) and the healed patients as a control group (n =260).By means of Logistic multivariate analysis,factors turned brucellosis into chronic were screened.Result The chronic brucellosis-related factors were:gender,veterinary or epidemic prevention staff,muscle and joint pain,fatigue symptoms,and substandard treatment (x2 =5.163,16.445,14.977,17.154,8.813,all P ＜ 0.05).Conclusion Gender (female),veterinary or epidemic prevention staff,muscle and joint pain,fatigue symptoms,and substandard treatment are probably the chronic brucellosis-related factors

Alzheimer’ s disease ( AD) , the most prevalent type of neurodegenerative disease, characterized by extracellular β-amyloid ( Aβ) plaque deposition, intracellular neurofibrillary tangles, and the loss of diverse neurons. Neuron loss might be the main cause that induces irreversible decline of cognitive function in AD patients. At present, AD therapy only relieves symptoms instead of fundamentally affecting the major pathologi-cal characteristics of the disease. Recently, rapid advances in neurogenesis and stem-cell biology have provided a new and pro-spective potential for AD treatment. Stem-cell types in the treat-ment of neurodegenerative diseases include neural stem cells ( NSCs ) , embryonic stem cells ( ESCs ) , mesenchymal stem cells ( MSCs ) , and induced pluripotent stem cells ( iPSCs ) . Stem cell therapy shows anti-AD function by replacing the dam-aged and lost neurons, rebuilding the cellular loop, inhibiting amyloidogenesis, promoting neurotrophic factor release, and reg-ulating immune reaction. This review highlights the recent pro-gress, mechanisms, and preclinical evidence of exogenous trans-planted stem cell, and addresses the current major challenges of stem cell transplantation in clinic. In addition, this review also summarizes the research status of compound promoting endoge-nous neurogenesis in brief.

Objective: To compare the effects of Cox proportional hazard regression model (Cox model) and extreme gradient boosting model ( XGBoost model ) on the prediction of the mortality of acute paraquat poisoning (APP). Methods: The APP cases admitted to Qingdao Eighth People's Hospital and Shandong Provincial Hospital from January 1st of 2018 to December 1st of 2020 was recruited and divided into a training group and a verification group by a random number table. The Cox model and XGBoost model were established to select the predictors for APP mortality. Receiver operating characteristic ( ROC ) curve was drawn to analyze the predictive power of the two models, and the calibration was evaluated using Hosmer-Lemeshow test. Results: Totally 150 APP cases were recruited. There were 75 cases each in the training group and in the verification group, with 52 and 55 cases died respectively, accounting for 69.33% and 73.33%. The Cox model showed that paraquat intake, the time from taking poison to seeing a doctor, the time for the first perfusion, the time for the first vomiting, aspartate aminotransferase, alanine aminotransferase, serum creatinine, blood urea nitrogen, white blood cell, lactic acid, creatine kinase isoenzymes, glucose, serum calcium and serum potassium were the predictors of APP mortality ( all P<0.05 ). The XGboost model showed that the predictive power of the factors in a descending order were the time from taking poison to seeing a doctor, the time for the first vomiting, the time for the first perfusion, lactic acid, white blood cell, paraquat intake, serum creatinine, serum potassium, serum calcium, creatine kinase isoenzymes, glucose, aspartate aminotransferase, blood urea nitrogen and alanine aminotransferase. The area under curve ( AUC ) of the XGBoost model for predicting was 0.972, which was greater than 0.921 of the Cox model ( P<0.05 ). The predicted results of the Cox model and XGBoost model were consistent with the actual situation ( P>0.05 ). Conclusion The Cox model and XGBoost model are consistent in predicting the mortality of APP, but the latter is better.

Objective To apply the microbial transformation to transforming the total saponins in the fruit of Panax ginseng (SFPG) and preparing ginsenoside compound K (C-K). Methods The four microbial strains m14, m3, m8, and m9 were screened and isolated from the soil in the botanic garden planted for P. ginseng and they were used for the microbial transformation of SFPG to optimize the strains. Taking C-K contents as index the microbial transformation was detected and analyzed by TLC and HPLC. Results The strain m14 was found to transform the SFPG efficiently to C-K at first. The optimal culturing and transformation conditions of m14 were obtained: time, 6 d; temperature, 30 ℃; revolution of cradle, 160 r/min; initial pH value, 5.5; substratum concentration, 120 mg/mL. Under the optimal condition, the content of C-K was 41.65 times as much as before transformation by m14. Conclusion The m14 is the most effective strain among the four fungal strains. It is the new way available for the C-K industrialized production.

Objective To study the mechanism of enhancing effect of pretreatment with Geqin Tablets (GT). Methods The rat preconditioning models of myocardial ischemia were established by an improved pushing tube method. Cell apoptosis was determined with TUNEL. The expression of PKC was examined by immunohistochemical method. The serum SOD activity was tested by hydroxylamine hydrochloride method and MDA activity by TBA method. Results The amount of apoptotic cells increased in ischemia /reperfusion (IR) group significantly(P