Cryptosporidium andersoni ATP-binding cassette (CaABC) is an important membrane protein involved in substrate transport across the membrane. In this research, the nucleotide binding domain (NBD) of CaABC gene was amplified by PCR, and the eukaryotic expression vector of pEGFP-C1-CaNBD was reconstructed. Then, the recombinant plasmid of pEGFP-C1-CaNBD was transformed into the mouse intestinal epithelial cells (IECs) to study the iron transportation function of CaABC. The results indicated that NBD region of CaABC gene can significantly elevate the transport efficiency of Ca2+, Mg2+, K+, and HCO3 - in IECs (P<0.05). The significance of this study is to find the ATPase inhibitors for NBD region of CaABC gene and to inhibit ATP binding and nutrient transport of CaABC transporter. Thus, C. andersoni will be killed by inhibition of nutrient uptake. This will open up a new way for treatment of cryptosporidiosis.
ATP-Binding Cassette Transporters/*chemistry/*genetics/metabolism ; Adenosine Triphosphate/metabolism ; Amino Acid Sequence ; Animals ; Calcium/metabolism ; *Cloning, Molecular ; Cryptosporidiosis/parasitology ; Cryptosporidium/chemistry/genetics/*metabolism ; Humans ; Iron/metabolism ; Mice ; Molecular Sequence Data ; Protein Structure, Tertiary ; Protozoan Proteins/*chemistry/*genetics/metabolism ; Sequence Alignment

OBJECTIVETo study the epidemiologic characteristics of hand-foot-mouth disease (HFMD) in Guiyang between 2008 and 2010.

METHODSThe epidemiologic characteristics of HFMD were analyzed by descriptive statistical methods based on the data from the China Information System for Disease Control and Prevention.

RESULTSA total of 27383 cases of HFMD were recorded in Guiyang between 2008 and 2010. The incidence of HFMD increased from 66.4439/100000 in 2008 to 163.9276/100000 in 2009 and 471.5515/100000 in 2010 (P<0.01). The mortality rate was 0.1026/100000 in 2010, which was significantly lower than in 2009 (0.2821/100000) (P<0.05). HFMD occurrence showed seasonality and reached a peak between April and June. HFMD cases were commonly noted in children under 5 years old, and especially in children under 3 years old. The main detected pathogen was human enterovirus 71 (EV17) in 2009. Whereas in 2010 the disease was mainly caused by CoxA16 and other intestinal viruses.

CONCLUSIONSThe incidence of HFMD in Guiyang increased year by year from 2008 to 2010, but the mortality rate decreased year by year. HFMD occurrence showed an obvious seasonality. HFMD was common in children under the age of five. The main pathogens of this disease included EV17, CoxA16 and other intestinal viruses.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Enterovirus A, Human ; isolation & purification ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans ; Infant ; Middle Aged ; Time Factors

Objective To study the dynamic air quality of clean ICU so as to provide evidence for hospital infection management in clean ICU.Methods Flat natural sedimentation method,six percussive determination of planktonic bacteria and dust particle counting method were used to get samples at three different time periods,different regions for four consecutive days in 100 000 air clean ICU.Results The counts of 0.5μm,5 μm dust particles at different time ( morning,afternoon,evening) were significantly different( F =78.85,89.94 ;P ＜ 0.01 ) and the monitor results of different regions( single rooms,double rooms,hall) were significantly different( F =20.21,16.17; P ＜ 0.01 ).The number of planktonic bacteria at different time (morning,afternoon,evening) were significant different(F =14.21,P ＜0.01 ),while there was no difference in different regions ( single rooms,double rooms,hall) ( F =0.98,P ＞ 0.05 ).There was significant difference of depositing bacterial counts at different time and regions( F =5.68,17.05,P ＜ 0.01 ) and there was a positively correlation between planktonic bacterial counts and depositing bacterial counts ( r =0.612,P ＜ 0.05 ).Each level of bacterial average counts of six percussive samplers measured was significantly different (F =8.35,P ＜ 0.01 ),with fifth grade most and fourth grade following.Conclusions Air quality of ICU is not good especially when making ward round; Less than 5 μm particles dominant; the counts of planktonic bacteria and depositing bacteria is increasing when dust particles counts increase,and the air quality of single rooms and double rooms is better than that of the hall.

Objective To investigate the effect of employers' respirator and hat wearing method on class 100 000 clean ICU air quality,in order to provide basis for hospital infection management in ICU.Methods For the control group,the first day (d1) and third day (d3),every employer was demanded to wear a respirator and a hat before entering the ICU.For the observation group,the second day (d2) and forth day (d4),employers were demanded not to wear any respirator or hat.During all four days,they must wear respirator and hat before any nursing operation.Air quality was sampled by class 100 000 clean air using flat panel natural settlement method,dust planktonic bacteria method and particle counting method.The amount of dust,plankton bacterium and descending bacteria were monitored for 4 days and compared between two groups.Results The difference of 0.5 μm dust particles in ICU between the control group and the observation group were statistically significant ( F =40.95,P ＜ 0.05 ).As to 5.0 μm dust particles,there was no significant difference between two groups (F =2.86,P ＞ 0.05 ).0.5 μm dust particles in ICU was lower at d1 and d3,and the difference was statistically significant ( F =40.95,P ＜ 0.05).The number of 0.5 μm dust particles had significant difference between different periods of a day:the morning time segmcnt＞ evening ＞ afternoon ( F =80.72,P ＜0.05 ).The number of 0.5 μm dust particles in the observation group was higher than that in the control group,and the difference was statistically significant ( F =68.84,P ＜ 0.05 ).The number of 5.0 μm dust particles had significant difference between different time periods of a day:the morning time segment ＞afternoon ＞ evening ( F =98.17,P ＜ 0.01 ).The number of dust particles at hall was larger than that at single and double rooms (P ＜0.01 ),but the number had no difference between single room and double room.More subsidence bacteria and floatingbacteria was detected during dl and d3 in the control group,and the difference was statistically significant ( P ＜ 0.01 ).Besides,the difference of the number of subsidence bacteria and floating bacteria in ICU between different time periods was also statistically significant ( P ＜ 0.05 ).Conclusions The number of 0.5 μm dust particles in ICU is smaller when workers wear hats and respirators,while the number of 5.0 μm dust particles remains the same.Under the other conditions remain unchanged,the mainly cause of number increase of dusts,planktonic bacteria and sedimentation bacteria in the morning rounds is the increasing number of working staff.ICU staff has no apparent effect on dynamic class 100 000 clean ICU air quality whether they wear respirators or hats.This kind of behavior is not the key management elements in clean ICU environment.

OBJECTIVEThis study aims to establish and evaluate the methodology of isolated rabbit eye (IRE) test.

METHODSIRE test was performed according to modifications of the in vitro toxicology (INVITTOX) Protocol No.85: Rabbit enucleated eye test by European Centre for the Validation of Alternative Methods (ECVAM), and then 26 chemicals and 26 cosmetic products were tested in both in vitro IRE and in vivo Draize tests. A statistical analysis was conducted to determine the relevance of the IRE test to the data generated in the Draize test.

RESULTSIRE test was established successfully in our laboratory. It was shown that ranking correlation and class concordance were fairly well between the IRE test and the Draize test for 26 reference chemicals (Fisher's Exact Test χ(2)=51.314, P<0.001; McNemar P=0.261; Gamma=0.960, P<0.001; Kappa=0.843, P<0.001) and 26 cosmetic products (Fisher's Exact Test χ(2)=15.522, P<0.001; McNemar P=0.311; Gamma=0.967, P<0.001; Kappa=0.611, P<0.001).

CONCLUSIONIRE test was established successfully for in vitro testing of eye irritation as an alternative to Draize test.

Animal Testing Alternatives ; Animals ; Cosmetics ; toxicity ; Eye ; drug effects ; Irritants ; toxicity ; Rabbits ; Toxicity Tests ; methods

Objective To investigate the effect of hairy and enhancer of split 5(HES5)on transdifferentiation and apoptosis of renal tubular epithelial cells induced by TGF-β1 and its potential mechanism.Methods The differentially expressed genes in GSE66494 data were analyzed and screened.The mouse model of unilateral uretera obstruction(UUO)was established,and the expression level of HES5 was detected in the renal tissue.HK-2 cells were treated with 10 ng/mL TGF-β1 for 24 h to establish a tubular epithelial-mesenchymal transition(EMT)model,and then qRT-PCR and Western blotting were performed to detect the expression of HES5 at mRNA and protein levels.After HK-2 cells were transfected with the plasmid overexpressing HES5,the protein levels of fibronectin,collagen Ⅰ,vimentin,apoptosis markers Bax and Bcl2 were detected in 24 h later.Then,HK-2 cells were divided into Control group,siHES5 group,TGF-β 1 group,and siHES5+TGF-β1 group.The protein level of fibrosis and apoptosis markers were measured in above groups with Western blotting.TUNEL staining and flow cytometry were employed to detect cell apoptosis.Western blotting was applied to determine the protein levels of AKT,p-AKT,PI3K and p-PI3K.HK-2 cells overexpressing HES5 were treated with PI3K inhibitor LY294002,and the expression of vimentin was detected.Results The expression of HES5 was significantly up-regulated in both chronic kidney disease(CKD)and fibrotic kidneys of mice.Overexpression of HES5 promoted the synthesis of fibronectin,collagen Ⅰ,vimentin and Bax in HK-2 cells,and inhibited the expression of Bcl2(P＜0.05).HES5 knockdown not only down-regulated the expression of fibrosis markers,but also inhibited the apoptosis of HK-2 cells.Furthermore,HES5 knockdown inhibited the activation of PI3K/AKT signaling pathway induced by TGF-β1 in HK-2 cells(P＜0.05).Inhibitors of the PI3K/AKT signaling pathway inhibitor attenuated the induction of HES5 on vimentin.Conclusion HES5 knockdown inhibits the transdifferentiation and apoptosis in TGF-β1-induced renal tubular epithelial cells,which may be related to the decreased activity of the PI3K/AKT signaling pathway.

OBJECTIVETo explore the correlation between single nucleotide polymorphisms (SNPs) of interleukin-28B (IL-28B) gene and the susceptibility to primary hepatocellular carcinoma (HCC).

METHODSA total of 300 histologically confirmed HCC cases (from November 2001 to April 2010) and 310 healthy controls with no history of chronic hepatitis B or hepatocellular carcinoma (2009-2010) were selected from a hospital in Guilin and a hospital in Beijing for this case-control study.139 HCC patients in the case group had complete clinical tracking data. All the subjects were Han Chinese, with no age or gender restrictions.2 ml peripheral blood samples were drawn from each subject with informed consent. SNP of rs12972991, rs4803223, rs8099917 and rs12979860 four loci in IL-28B gene were analyzed by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF).

RESULTSThe frequencies of C allele at rs12972991, G allele at rs8099917 and G allele at rs4803223 were 6.7% (40/598), 7.9% (47/598) and 10.0% (59/588) respectively in case group; all higher than the corresponding frequencies in control group, separately 2.9% (18/618), 4.1% (25/616) and 3.6% (21/608). The differences were statistically significant (χ2=9.542, 7.858, 20.736, P values all<0.05). The above alleles could increase the risk of HCC, and the OR (95%CI) values were separately 1.67 (1.13-2.46), 1.49 (1.08-2.06) and 2.91 (1.79-4.72). The genotype frequencies of AC+CC at rs12972991, GT+GG at rs8099917, GA+GG at rs4803223 were 13.0% (39/299), 14.7% (44/299) and 19.0% (56/296) respectively in case group; while the frequencies were lower in control group, separately 5.8% (18/309), 8.1% (25/308) and 6.6% (20/304). The differences were statistically significant (χ2=9.319, 6.557, 20.948, P values all<0.05). These genotypes may increase the risk of HCC, and the adjusted OR (95%CI) values were 2.24 (1.31-3.83), 1.81 (1.14-2.88) and 2.90 (1.78-4.70), respectively. The stratified analysis of the clinical data indicated that the frequency of genotype GA+GG at rs4803223 was 50.0% (13/26) in patients of tumor thrombosis in portal vein (TTPV), higher than the frequency of genotype AA (21.1%, 23/109). The difference was statistically significant (χ2=8.965, P=0.003).

CONCLUSIONThe results suggested that IL-28B gene polymorphisms was correlated to the susceptibility to HCC in Chinese Han ethnic population. Among them, GA + GG genotype at rs4803223 could increase the risk of TTPV in HCC patients.

Alleles ; Carcinoma, Hepatocellular ; genetics ; Case-Control Studies ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Interleukins ; genetics ; Liver Neoplasms ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide

Scutellarin is a flavonoid extracted from breviscapus， a traditional Chinese medicine. Pharmacological studies have shown that scutellarin has anti-inflammatory， antioxidant， anti-fibrosis， anti-tumor， improving cardiac and cerebral ischemia. In recent years， with the deepening of research on scutellarin， it was found that it could inhibit the tumor through multi-target and multi-pathway， and the anti-human colorectal cancer was related to the regulation of p53 pathway， Hedgelog pathway and erythropoietin generates liver cancer interactivator B2（EphrinB2）.The anti-esophageal squamous cell carcinoma is related to protein kinaseB1 /protein kinaseB2（ Akt1/Akt2）.Anti-renal carcinoma and melanoma are associated with phosphatase and tension protein homologues（PTEN） and phosphatidylinositol 3-kinase（PI3K）/Akt/mammalian target of rapamycin（mTOR） pathway. Anti-lung cancer is related to Akt/mTOR/4E binding protein1（4EBP1） and signal transduction and transcriptional activator（STAT3 ）signaling pathway. Anti-cervical cancer is related to pyruvate kinase 2（PKM2）.Anti-breast cancer is associated with Hippo/YAP pathway. At the same time， scutellarin was found to prevent diabetic microangiopathy， atherosclerosis and non-alcoholic fatty liver disease by regulating glucose and lipid metabolism， but the mechanism of action was not well studied. A review of the literature found that scutellarin anti-tumor， atherosclerosis， diabetic microangiopathy， non-alcoholic fatty liver disease， osteoarthritis， osteoporosis mechanism of action lack of detailed summary. In this paper， the research progress of pharmacological action and mechanism of scutellarin in recent 5 years is reviewed， and Suggestions on its current research status and future direction are put forward， in order to speed up the discovery of pharmacological mechanism of scutellarin and provide scientific basis for its further development and utilization.

Objective: To analyze the change trend of underground dust concentration, the incidence and survival status of pneumoconiosis patients, and provide reference for improving the working environment of dust-exposed workers and the prevention and treatment of pneumoconiosis patients in the future. Methods: In February 2020, a retrospective investigation was conducted on the dust data of underground mining operations in a steel enterprise in Jiangsu Province from 1991 to 2019, and the case data of patients diagnosed with pneumoconiosis from 1956 to 2019 were collected. The time trends of the number of pneumoconiosis patients and dust concentration, the stage of pneumoconiosis and survival status of patients were analyzed. Results: From 1956 to 2019, a total of 241 patients with pneumoconiosis were diagnosed in the steel enterprise. From 1991 to 2019, the annual average dust concentration in the mine showed a downward trend as a whole. Compared with the transportation platform (14.28%, 1447/10132) , the average dust concentration exceeding rate of the mining platform (43.68%, 8415/19263) was significantly higher (χ(2)=2674.84, P<0.01) . The average age of pneumoconiosis patients was (73.54±10.42) years old, and the average working age of dust exposure was (21.41±8.68) years, of which 85 cases (35.27%) survived and 156 cases (64.73%) died. The main type of pneumoconiosis was silicosis (90.46%, 218/241) , and the main stage of pneumoconiosis was the stage I (96.68%, 233/241) ; The higher the stage of pneumoconiosis, the younger the diagnosis age (P<0.01) . The average survival time of patients was (27.264±1.982) years, and the median survival time was 28 years. The cumulative survival rates of patients with pneumoconiosis in different diagnosis time periods were significantly different (χ(2)=35.57, P<0.01) . Conclusion: The improved dust-proof measures have a significant effect on reducing the concentration of underground dust. We need to focus on the dust control of underground mining platforms and the treatment of patients with stage Ⅲ pneumoconiosis.
Humans ; Middle Aged ; Aged ; Aged, 80 and over ; Child ; Adolescent ; Young Adult ; Adult ; Incidence ; Coal Mining ; Steel ; Retrospective Studies ; Pneumoconiosis/diagnosis* ; Dust ; Occupational Exposure/adverse effects*

Objective: To establish a method for the determination of acetylacetone in the air of workplace by gas chromatography. Methods: In August 2020, acetylacetone in the air of workplace was collected by silica gel tube, eluted with methanol, separated and detected by gas chromatography with flame ionization detector. The detection limit and precision of the method were also analyzed. Results: The linear range of acetylacetone was 1.95-1950.60 μg/ml with the regression equation of y=0.815x-3.667, and the correlation coefficient was 0.99993. The limit of detection of the method was 0.18 μg/ml and the minimum detection concentration was 0.12 mg/m(3) (collected sample volume was 1.50 L). The within-run precisions were 1.08%-4.11% and the between-run precisions were 1.98%-2.80%. The desorption rates were 99.68%-100.45%. The sealed samples could be kept at least 15 days at room temperature without significant loss. Conclusion: The solvent desorption-gas chromatography method for the determination of acetylacetone has good precision, high sensitivity and simple operation, and is suitable for the determination of acetylacetone in the air of the workplace.
Solvents ; Air Pollutants, Occupational/analysis* ; Chromatography, Gas/methods* ; Workplace