With reverse transcriptase PCR, the transcripton of copper homeostasis relative gene Afe0329 in Acidithiobacillus ferrooxians standard strain ATCC23270 was investigated. The further analysis of genes in this transcripton was analyzed employed by Vector NTI, Blast, TMHMM Server, PSORTb software and so on. From the DNA of different strains, the transcripton of Afe0329 was amplified using special primer pairs to identify the universality of it in the genome of A.ferrooxidans strains. The results showed that gene Afe0330 and Afe0331 were cotranscribed with Afe0329, and they were in a single transcripton. Gene Afe0329 was supported to express a P1b3-type ATPase which is a heavy metal ion pumping transmembrane protein, protein AFE0330 which expressed by gene Afe0330 was a cytoplasmic protein, no significant ho- mologous sequences of Afe0330 or Afe0331 had been obtained by Blast analysis. And the transcripton of Afe0329 was universal in genome of A. ferrooxidans strains.

Objective To explore the regulation of ROS level and ROS-triggered downstream events on SK-N-MC Ewing sarcoma cells upon apoptasis induction by 2-Methoxyestradiol (2-ME). Methods To detect the reversibility of apoptosis and the alternation of activity of respiratory chain, mitechondria transmembrane potential (△ψm), and cellular ROS level and to explore their association with flow cytometry, clark oxygen electronic node analysis, drug-removal design, and permeability transition (PT) pore stablizing agent. Results SK-N-MC cells were induced to ROS-dependent apoptosis. Apoptosis occured irreversibly after2-ME treatment for 3 h. Upon 2-ME treatment, the activity of respiratory chain was inhibited and the ROS generation was accelerated; the △ψm underwent the increasing within 3h but decreasing after 3h which could be reversed by PT pore stablizing; the ROS level underwent the continuous increasing and PT pore stablizing had no obvious effect on it. Conclusion 2-ME causes the acceleration of ROS generation via inhibiting the activity of respiratory chain and elevating the level of △ψm. ROS plays a signaling role and when total ROS accumulate to a threshold, the PT pore opening and the collapse of △ψm could be induced irreversibly and cell is eventually introduced to death.

OBJECTIVE: To observe the protective effect of Huaxia shallot preparation on human umbilical vein endothelial cell (HUVEC) injury induced by oxidized low density lipoprotein (Ox-LDL) in vitro. METHODS: Ox-LDL was prepared and identified, and HUVECs were cultured. After 2-hour intervention of different drugs and 24-hour following intervention of Ox-LDL, the number of HUVECs was observed by phase contrast optical microscope and the activity of the HUVECs was observed by methyl thiazolyl tetrazolium (MTT) technique. Superoxide dismutase (SOD) activity and nitric oxide (NO) content were assayed by respective kit. The protein expressions and mRNA levels of peroxisome proliferators activated receptor gamma(PPAR-gamma) and endothelial nitric oxide synthase (eNOS) were measured by western blot technique and reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Ox-LDL could increase the apoptosis rate of the HUVECs and decrease the NO release as compared with the blank control group (P<0.05). These effects induced by Ox-LDL were all significantly inhibited by Huaxia shallot preparation. It could up-regulate the protein expressions and mRNA levels of PPAR-gamma and eNOS significantly (P<0.05). Huaxia shallot preparation could decrease the apoptosis rate of the HUVECs. CONCLUSION: Ox-LDL may be involved in the initiation and progression of atherosclerosis by injuring the endothelial cells directly and may cause the endothelial dysfunction. Huaxia shallot preparation can protect against Ox-LDL induced endothelial cell injury by up-regulating the protein expressions and mRNA levels of PPAR-gamma and eNOS. It suggests that Huaxia shallot preparation may play a role in the prevention and treatment of cardiovascular disease.

Mild cognitive impairment (MCI) is a pre-clinical stage of Alzheimer's disease (AD). In recent years, transcranial direct current stimulation (tDCS), as a neuromodulation technique, has being applied in the field of cognitive intervention for MCI, but its effect is controversial because of many factors. In order to promote the application of tDCS in intervention for MCI, this study performed a systematic review of the previous studies that used tDCS to improve cognitive functions of MCI individuals. The results indicate that tDCS could improve episodic memory, working memory, and language of individuals with MCI, while there is a lack of strong evidence that supports a positive effect of tDCS on attention of individuals with MCI. The placement of electrodes, time course of treatment, and current intensity all affected the intervention effect of tDCS. Future studies should take brain networks underlying cognitive processes and personalized factors such as age and education level into consideration to design better stimulation protocols, and they should be conducted in combination with neuroimaging technologies to evaluate the intervention effect of tDCS more accurately and objectively and to discover the neural mechanisms of tDCS intervention.

Objective : To explore the relationship between alcohol consumption and hypertension in adults of Zhejiang Province, so as to provide scientific evidence for the prevention and control of hypertension. Methods: Data were collected from 10 national surveillance sites in Zhejiang Province in the 2018 China Chronic Disease and Risk Factors Surveillance. The multivariate logistic regression model was used to explore the relationship between alcohol consumption and hypertension in adults of Zhejiang Province. Results : Among 5 908 people included, 2 641 were males, accounting for 50.38%; and 3 267 were females, accounting for 49.62%. There were 2 721 cases of hypertension ( 28.54% ). The number of the cases with no, light and excessive alcohol consumption was 3 945, 1 117 and 846, accounting for 66.28%, 23.65% and 10.07%, respectively, after complex sampling weighting. The results of the multivariate logistic regression model showed that compared with those with no alcohol consumption, the adults with excessive alcohol consumption had higher risk of hypertension ( OR=2.126, 95%CI: 1.649-2.741 ); men with light ( OR=1.875, 95%CI: 1.075-3.270 ) and excessive alcohol consumption ( OR=2.752, 95%CI: 2.021-3.748 ) had higher risk of hypertension; the 18-<45-year-old adults with light ( OR=2.441, 95%CI: 1.171-5.087 ) and excessive alcohol consumption ( OR=3.368, 95%CI: 1.609-7.049 ), and the ≥60-year-old adults with excessive alcohol consumption ( OR=1.632, 95%CI: 1.174-2.269 ) had higher risk of hypertension. Conclusions Light and excessive alcohol consumption are associated with hypertension in Zhejiang adults. The association is stronger between excessive alcohol consumption and hypertension than between light alcohol consumption and hypertension.

OBJECTIVETo explore the expression of Survivin (SVV) protein in colorectal carcinogenesis and its clinical significance. METHODS Immunohistochemistry staining was performed by two-step EnVision technique for the paraffin sections, which included 90 adenomas, 25 ademomas with high-grade glandular intraepithelial neoplasia, and 108 colorectal adenocarcinomas.

RESULTSExpressions of SVV, P53, and Bcl-2 were observed in tumor cells of the sections. The positive rate of SVV in tubular adenomas, villous adenomas, and tubulovillous adenomas were 30% (12/40), 40.9% (9/22), and 35.8% (10/28), respectively. The positive rate of SVV in tubulovillous adenomas with high-grade glandular intraepithelial neoplasia were 68% (17/25). The positive rate of SVV in carcinomas of stage A, B, and C were 75% (27/36), 81.3% (26/ 32), and 95% (38/40), respectively. SVV expressions among the three types of adenomas without neoplasia were not significantly different (P > 0.05). SVV expression between each type of the above-mentioned ademoma and tubulovillous adenoma with high-grade glandular intraepithelial neoplasia or different Dukes stages of colorectal carcinoma was significantly different (P < 0.05). SVV expressions in adenocarcinomas and adenomas with high grade glandular intraepithelial neoplasia were significantly higher than those in adenomas (P < 0.01). The expressions of P53 and Bcl-2 had no significant difference among them. No association was noted between SVV expression and P53 or Bcl-2 expression (P = 0.487, P = 0. 437).

CONCLUSIONSSVV is abnormally expressed in the early stage of colorectal carcinogenesis, which may be correlated with the carcinogenesis of colorectal ademoma. SVV expression may be useful to distinguish adenocarcinoma from adenoma in colorectal carcinogenesis.

Adenocarcinoma ; metabolism ; pathology ; Adenoma ; metabolism ; pathology ; Colorectal Neoplasms ; metabolism ; pathology ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; biosynthesis

OBJECTIVETo observe the therapeutic effect of local antibiotic injection into the female prostate on female urethral syndrome (FUS), and search for an effective treatment for this disease.

METHODSThis study included 163 FUS patients treated in the out-patient department between July 2009 and December 2010. According to the visiting order, the patients were randomly assigned to Groups A (n = 58), B (n = 55) and C (n = 50). All underwent routine treatment. Inaddition Group A received local injection of 2 ml of 80 000 U gentamycin + 2 ml of lidocaine, and Group B 2 ml of normal saline + 2 ml of lidocaine, both injected into the distal segment of the urethral back wall where the female prostate is located, twice a week for 3 weeks. The therapeutic effects were evaluated according to the changes of the patients' independent symptom scores at 2 and 4 weeks after the treatment. Disappearance of the symptoms was considered as "curative" , > 1/2 reduction in the symptom score as "obviously effective", 1/2 - > 1/4 reduction in the symptom score as "effective", and < 1/4 reduction or increase in the symptom score as "ineffective".

RESULTSAt 2 weeks after the treatment, the total effectiveness rate was significantly higher in Group A (77.5%) than in B (67.3%) and C (68.0%) (P < 0.05), but with no statistically significant difference between B and C (P > 0.05). At 4 weeks, the total effectiveness rate of Group A was slightly decreased, but still remarkably higher than that of group B or C (P < 0.05).

CONCLUSIONLocal injection of gentamycin into the female prostate is effective for the treatment of female urethral syndrome.

Administration, Topical ; Adult ; Aged ; Aged, 80 and over ; Female ; Gentamicins ; administration & dosage ; therapeutic use ; Humans ; Injections ; Middle Aged ; Prospective Studies ; Treatment Outcome ; Urethral Diseases ; drug therapy ; Young Adult

OBJECTIVETo explore the influence of different thawing temperatures on the morphology and type I collagen metabolism of the human fibroblasts processed at - 10 degrees C in vitro.

METHODSIn vitro cultured human fibroblasts were randomly divided into control, 20 degrees C thawing, and 37 degrees C thawing groups. After being frozen at -10 degrees C, the cells in the latter two groups were thawed at 20 degrees C and 37 degrees C, respectively. The cell proliferation was assessed with MTT method and was expressed by absorption under 570nm (A ), The morphological change of the cells was observed with inverted phase contrast microscope, the change in the intracellular content of collagen was determined with immunohistochemistry, and the extracellular content of collagen was assayed with ELISA.

RESULTSIn 20 degrees C thawing group, the absorbance decreased at first and increased thereafter, and they were obviously lower than that before freezing (0.95 +/- 0.16, P < 0.05 or 0.01). Cell dehydration and shrinking, cytoplasm loss and increased ratio of cytoplasm to nucleus were found in the survived fibroblasts. The cells proliferated actively at 72 and 96 hours after injury, with increased mitotic index and disordered arrangement. Compared with that before freezing (96.4 +/- 2.9) , the extracellular collagen content increased at first, decreased thereafter, and increased again slowly later (P < 0.05), while the intracellular collagen content decreased at first and increased thereafter (P < 0.05). The collagen metabolism in 37 degrees C thawing group was no difference compared with that in control group. Some cells undergone a floating period before adhering to the culture dish walls.

CONCLUSIONCell dehydration after low temperature treatment could protect the cells from damage. Proper thawing temperature could be beneficial to the cell resuscitation. Comparing with slow thawing, rapid thawing could minimize the cell damage.

Cells, Cultured ; Collagen Type I ; metabolism ; Cryopreservation ; Fibroblasts ; cytology ; metabolism ; Freezing ; Humans

BACKGROUNDThe Borg scale is most commonly used to measure dyspnea in China. However, many patients that find it is difficult to distinguish the labeled numbers corresponding to different dyspnea scores. We developed a new method to rate dyspnea, which we call the count scale (CS). It includes the count scale number (CSN) and count scale time (CST). The aims of the present study were to determine the reproducibility and sensitivity of the CS during exercise in patients with chronic obstructive pulmonary disease (COPD).

METHODSFourteen male patients with COPD (aged 58.00 ± 7.72 years) participated in this study. A progressive incremental exercise and a 6-minute constant work exercise test were performed every 2 to 3 days for a total of 3 times. The CS results were evaluated at rest and at 30% and 70% of maximal workload (Wmax) and Wmax. The Borg scales were obtained during exercise.

RESULTSNo significant differences occurred across the three trials during exercise for the CS and Borg scores. The CSN and CST were more varied at Wmax (coefficient of variation (CV) = (22.28 ± 16.96)% for CSN, CV = (23.08 ± 19.11)% for CST) compared to 30% of Wmax (CV = (11.92 ± 8.78)% for CSN, CV = (11.16 ± 9.96)% for CST) and 70% of Wmax (CV = (9.08 ± 7.09)% for CSN, CV = (12.19 ± 12.32)% for CST). Dyspnea ratings with either CSN or CST tended to decrease at the higher workload compared to the lower workload. CSN and CST scores were highly correlated (r = 0.861, P < 0.001). CSN was negatively correlated with Borg scores (r = -0.363, P = 0.001). Similar results were obtained for the relationship between CST and Borg scores (r = -0.345, P = 0.003).

CONCLUSIONWe concluded that the CS is simple and reproducible when measuring dyspnea during exercise in patients with COPD.

Aged ; Dyspnea ; diagnosis ; Exercise ; Humans ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; physiopathology ; Reproducibility of Results

Objective:To analyze the clinical characteristics and the risk factors of acute mechanical small bowel obstruction in adults.Methods:The clinical data of 487 adult patients with acute mechanical small bowel obstruction treated in Drum Tower Hospital Clinical College of Nanjing Medical University from June 2010 to December 2020 were retrospectively analyzed. There were 259 cases of strangulated obstruction (strangulation group) and 228 cases of non-strangulated obstruction (simple group). The cases in the strangulation group were confirmed by operation or pathology, the cases in the simple group were confirmed by non-operative therapy ( n=167) or operation ( n=61). The clinical data, including age, abdominal pain, vomiting, tenderness, rebound pain and muscle defense; the CT signs, including transitional zone, dilatation of intestinal loop, high density of intestinal wall; laboratory test results, including white blood cell count, lactate dehydrogenase (LDH) and C-reactive protein (CRP) were analyzed in both groups. Univariate and multivariate regression were used to identify risk factors of strangulated small bowel obstruction. Results:There were significant significances in gender (male vs.female: χ2=4.35, OR=0.67), rebound pain (χ2=170.98, OR=45.12), muscle tension(χ2=113.79, OR=29.32), American Society of Anesthesiologists score (ASA score)≥3 (χ2=12.94, OR=51.58), white blood cell count ( t=6.19, OR=1.14), LDH ( t=2.37, OR=1.00), CRP ( t=2.23, OR=1.01), albumin ( t=2.04, OR=0.97), mesenteric fluid sign (χ2=49.44, OR=5.40), increased bowel wall density (χ2=222.78, OR=62.66), bowel wall thickening sign (χ2=43.81, OR=3.49), ascites (χ2=237.29, OR=43.01), beak sign (χ2=231.50, OR=56.62), mesenteric fat stranding (χ2=242.65, OR=34.90), and stranding sign(χ2=224.79, OR=53.48) between strangulation group and simple group ( P<0.01). The multivariate regression analysis showed that mesenteric fluid sign ( OR=12.94), muscle tension ( OR=7.28), ascites ( OR=6.42), increased bowel wall density ( OR=4.30), bowel wall thickening sign ( OR=1.85), white blood cell count ( OR=1.14) and gender (male vs. female: OR=0.50) were risk factors of strangulated small bowel obstruction. Conclusion:In acute mechanical small bowel obstruction,for female patients presenting mesenteric fluid sign, muscle tension, ascites, increased bowel wall density, bowel wall thickening sign and increased white blood cell count, the strangulated obstruction is likely to occur.