An effective RT-PCR method was developed to detect exogenous gene xylB transcript levels in Zymomonas mobilis CP4. Total RNAs without genomic DNA contamination were purified from wild-type and gene engineering strains, and were quantified to the same concentration. Then, cDNAs synthesis and PCR analysis of these samples were conducted by reverse transcription PCR. The optimal number of cycles was determined by observing amplification profile of target gene xylB and internal control gene 16s rRNA, and relative expression levels of xylB in various samples were analyzed by RT-PCR. The results indicated that the xylB transcript was not be detected in CP4, however that could be found in recombinant strains, in which xylB transcription abundance was similar. The enzyme assay furthermore confirmed that effective ex-pression of the target gene. The method provided a useful and rapid tool for detecting transcript levels of target genes from various samples of Z. mobilis.

OBJECTIVETo detect the toxicity and teratogenicity of 2, 2-dinitroethene-1, 1-diamine (FOX-7) in rats.

METHODS125 adult SD rats were randomly divided into five groups, which are negative control (0 mg/kg) , positive control (280 mg/kg aspirin) , and three dose groups (5, 15, and 45 mg/kg) . They were administrated by gavage once a day from the 5th days to 19th days after pregnancy. The weight changes and toxicity of pregnant rats are recorded within the study, and the skeleton and internal organs malformations are detected by the recommended methods.

RESULTSAfter 5 or 6 days being poisoned, the pregnant rats appear significantly toxicity symptoms, such as exciting, irritability, and so on. The net weight raise in high dose group is less than the negative group, while the numbers of dead foetus in median and high dose groups are both more than that of negative group. Comparing with the negative group, the body weight and body lenghth of foetus rats in median and high dose groups, and the tail lenghth in high dose group are lower significantly. There are no external malformations in negative group and three dose groups. However, the foetus of high dose group appear significant skeleton and internal organs malformation prevalences that are significant more than negative group, including lateral cerebral ventricles enlarged, which accounts for 9.17%, occipital bone lost, which accounts for 2.59%.

CONCLUSIONFOX-7 can induced maternal reproductive toxicity, foetus toxicity and teratogenicity hazards to rats.

Animals ; Body Weight ; Female ; Nitro Compounds ; toxicity ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Teratogens ; toxicity ; Toxicity Tests

Objective:To prepare loratadine nanosuspension lyophilized powder (LTD-NLP) and investigate the dissolution and stability. Methods:LTD-NLP was prepared by an anti-solvent precipitation method; the particle size and morphology were characterized by a laser nanometer particle sizer and a transmission electron microscope (TEM). The optimal lyophilized protective agent was screened out; the dissolution and stability of LTD-NLP were determined by HPLC. Results:The average particle size of LTD-NLP was 175.6 nm with PDI of 0.200, and the zeta potential was -56.3 mV. The shape of LTD-NLP was spherical. The nanosuspension lyophilized with 10% sucrose presented optimal properties. The dissolution of LTD-NLP was greater than raw material, and LTD-NLP had good stability at 4℃. Conclusion:The anti-solvent precipitation method for LTD-NLP is simple,and the product is expected to become a new preparation of loratadine.

Objective: To construct the cell models of TNF-α rs1800620 (SNP1) and TNF-α rs4645843 (SNP2) stable transgen-ic lines and study the effects of SNPs on the content of TNF-α in LX-2 cell line. Methods: TNF-α SNP1/TNF-α SNP2 mutant plasmid was synthesized by wild-type TNF-α targeting and then transfected into LX-2 cell line after lentivirus packaging. RT-qPCR and Western blot were used to detect the mRNA and protein levels of TNF-α in each recombinant cell. Results: The gene sequencing showed that plasmids were successfully constructed. Photographs from an inverted microscope showed that the recombinant cells had stable expres-sion of GFP. The results of RT-qPCR and Western blot showed that compared with the wild group, SNP1 and SNP2 could significantly reduce the mRNA (P<0. 05) and protein (P<0. 05) levels of TNF-α. Conclusion: The recombinant cells stably expressing TNF-α/TNF-α SNP1/TNF-α SNP2 LX-2 are constructed. SNP1 and SNP2 can significantly reduce the mRNA and protein levels of TNF-α in LX-2 cell line.

Objective To evaluate the value of color Doppler two-dimensional ultrasound combined with real-time ultrasonic shear wave elastography (SWE) in the early diagnosis of breast cancer.Methods A total of 110 suspected breast tumor patients from October 2015 to April 2017 were selected as subjects.Comprehensive detailed examinations on the patients were conducted by routine two-dimensional ultrasound,color Doppler and SWE,and the lesion location,size,boundary,capsule,internal echo,blood flow,elastic value and other related data were recorded.The BI-RADS classification was conducted based on these data.Pathological diagnosis was used as the gold standard to compare the diagnostic effects of color Doppler two-dimensional ultrasound and the combination of color Doppler two-dimensional ultrasound and SWE.Results Of the 110 patients,78 were malignant and 32 were benign.The sensitivity of color Doppler two-dimensional ultrasound for breast cancer diagnosis was 83.3％,the specificity was 81.2％,and the accuracy was 82.7％.The sensitivity of color Doppler two-dimensional ultrasound combined with SWE for breast cancer diagnosis was 88.5％,the specificity was 90.6％,and the accuracy was 94.5％.Conclusion Color Doppler two-dimensional ultrasound combined with SWE can complement each other and improve the qualitative and quantitative diagnosis of early breast cancer.

OBJECTIVETo investigate whether 3,4-methylenedioxymethamphetamine (MDMA) abuse produces another neurotoxicity which may significantly inhibit the acetylcholinesterase activity and result in severe oxidative damage and liperoxidative damage to MDMA abusers.

METHODS120 MDMA abusers (MA) and 120 healthy volunteers (HV) were enrolled in an independent sample control design, in which the levels of lipoperoxide (LPO) in plasma and erythrocytes as well as the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and acetylcholinesterase (AChE) in erythrocytes were determined by spectrophotometric methods.

RESULTSCompared with the average values of biochemical parameters in the HV group, those of LPO in plasma and erythrocytes in the MA group were significantly increased (P < 0.0001), while those of SOD, CAT, GPX and AChE in erythrocytes in the MA group were significantly decreased (P < 0.0001). The Pearson product-moment correlation analysis between the values of AChE and biochemical parameters in 120 MDMA abusers showed that significant linear negative correlation was present between the activity of AChE and the levels of LPO in plasma and erythrocytes (P < 0.0005-0.0001), while significant linear positive correlation was observed between the activity of AchE and the activities of SOD, CAT and GPX (P < 0.0001). The reliability analysis for the above biochemical parameters reflecting oxidative and lipoperoxidative damages in MDMA abusers suggested that the reliability coefficient (alpha) was 0.8124, and that the standardized item alpha was 0.9453.

CONCLUSIONThe findings in the present study suggest that MDMA abuse can induce another neurotoxicity that significantly inhibits acetylcholinesterase activity and aggravates a series of free radical chain reactions and oxidative stress in the bodies of MDMA abusers, thereby resulting in severe neural, oxidative and lipoperoxidative damages in MDMA abusers.

Acetylcholinesterase ; metabolism ; Adolescent ; Adult ; Amphetamine-Related Disorders ; blood ; enzymology ; metabolism ; Catalase ; blood ; Cholinesterase Inhibitors ; adverse effects ; urine ; Erythrocytes ; enzymology ; Female ; Humans ; Lipid Peroxidation ; drug effects ; Lipid Peroxides ; blood ; Male ; N-Methyl-3,4-methylenedioxyamphetamine ; adverse effects ; urine ; Oxidative Stress ; drug effects ; Superoxide Dismutase ; blood

OBJECTIVETo investigate whether hypertension, abnormal lipometabolism, obesity, cigarette smoking and alcohol drinking affect the intracerebral hemorrhagic volumes (IHV) in patients with spontaneous intracerebral hemorrhage (SIHP), and to explore the roles of these factors in spontaneous intracerebral hemorrhage (SIH).

METHODSFive hundred patients with acute SIH and 200 healthy adult volunteers (HAV) were enrolled in a study of independently randomized controlled design, in which the levels of systolic pressure (SP) and diastolic pressure (DP), and total cholesterol (TCH), triacylglycerols (triglycerides, TG), high density lipoprotein cholesterol (HDL-CH), low density lipoprotein cholesterol (LDL-CH) in serum as well as the level of erythrocytic membrane cholesterol (EM-CH) were measured, and the body mass index (BMI), daily cigarette smoking consumption (DCSC) and daily pure alcohol consumption (DPAC) were calculated.

RESULTSCompared with the average parameters in the HAV group, those of SP, DP, TG, LDL-CH and BMI in the SIHP group were significantly increased (P < 0.0001), while those of HDL-CH and EM-CH were significantly decreased (P < 0.0001). The linear regression and correlation analysis showed that with increased SP, DP, LDL-CH, BMI, DCSC, DPAC and aging as well as decreased HDL-CH and EM-CH, the IHV levels in SIHP were increased gradually (P < 0.0001-0.01). The linear stepwise regression analysis suggested that there existed a close correlation among the values of SP, DP, TCH, TG, HDL-CH, LDL-CH, EM-CH, BMI, DCSC, DPAC, age and IHV of the SIH patients, and that Y = -12.4583 + 0.1127SP -1.1977EM-CH + 0.9788LDL-CH + 0.2477BMI + 0.0382DCSC + 0.0248DP, P < 0.0001 approximately 0.05.

CONCLUSIONSThe findings in the present study suggest that significantly increased systolic and diastolic pressure, low density lipoprotein cholesterol, body mass index and daily cigarette smoking consumption, and significantly decreased erythrocytic membrane cholesterol may be likely the main factors affecting intracerebral hemorrhagic volumes in patients with acute spontaneous intracerebral hemorrhage.

Aged ; Blood Pressure ; Body Mass Index ; Case-Control Studies ; Cerebral Hemorrhage ; etiology ; Cholesterol, LDL ; blood ; Female ; Humans ; Hyperlipidemias ; complications ; Hypertension ; complications ; Life Style ; Male ; Middle Aged ; Obesity ; complications ; Regression Analysis ; Risk Factors ; Smoking ; adverse effects

OBJECTIVE: To evaluate the prospective association between cumulative resting heart rate (cumRHR) and rapid renal function decline (RRFD) in a cohort of individuals aged 60 and older. METHODS: In the Tianjin Chronic Kidney Disease Cohort Study, the individuals who underwent three consecutive physical examinations between 2014 and 2017, with estimated glomerular filtration rate (eGFR) greater than 60 mL/min per 1.73 m² and aged 60 years or older were enrolled. A total of 27,564 patients were prospectively followed up from January 1, 2017 to December 31, 2020. The 3-year cumRHR was calculated. The primary outcome was RRFD, defined as an annualized decline in eGFR of 5 mL/min per 1.73 m² or greater. Logistic and restricted spline regression models and subgroup analysis were used to investigate the association of cumRHR with RRFD after adjusting for all confounders. RESULTS: During a median follow-up of 3.2 years, a total of 4,347 (15.77%) subjects developed RRFD. In fully-adjusted models, compared with the lowest quartile of cumRHR, the odds ratio (OR) for the highest was 1.44 (1.28-1.61), P < 0.001. Furthermore, each 1-standard deviation (27.97 beats/min per year) increment in cumRHR was associated with a 17% (P < 0.001) increased risk of RRFD, with a linear positive correlation (P for non-linear = 0.803). Participants with a 3-year cumRHR ≥ 207 (beats/min) * year (equivalent to ≥ 69 beats/min per year in 3 years) were found to be at a higher risk of RRFD. CONCLUSIONS The cumRHR is significantly associated with a higher risk of RRFD among older adults. These results might provide an effective goal for managing and delaying the decline of renal function in the older adults.