1.Genotyping of candida glabrata clinical isolates by repetitive sequence-based PCR and multilocus sequence typing
Danfeng DONG ; Cen JIANG ; Beiqin YU ; Qishi FAN ; Yibing PENG
Chinese Journal of Laboratory Medicine 2011;34(9):810-813
Objective To assess the application value of REP-PCR in genotyping of candida glabrata strains in clinical pratice. MethodsFrom 2009 to 2010, thirty-eight candida glabrata strains were isolated from Shanghai Ruijin Hospitals, Shanghai Renji Hospital, Shanghai Huashan Hospital, Anhui Medical University Hospital, Shenzhen People's Hospital. Six loci in housekeeping genes (FKS, LEU2,NMT1, TRP1, UGP1 and URA3 ) were amplified and sequenced. The sequences were compared with the MIST database and allele profile and sequence type (ST) were obtained. With primers Ca21, Ca22 and Com21 used to amplify the adjacent variable gene regions,the amplicons were analyzed through electrophoresis to generate different REP-PCR types. Finally, the results of these two genotyping methods were compared. ResultsFor REP-PCR, Ca22-Com21 has the best genotyping effect. REP-PCR and MLST have the same genotyping results. Five REP-PCR types were found in 38 candida glabrsta isolates. Type A,B, C, D and E strains from REP-PCR were genotyped as ST 7, 3, 19, 45 and new type respectively byMIST. REP-PCR saves time compared with MIST. Conclusions REP-PCR offers a simple and rapid method for molecular typing, with similar discriminatory power with MIST. Therefore, REP-PCR can be the preferred choice in laboratory, especially for a large number of isolates.
2.Effects of tobacco extract on the proliferation and HSP70 expression of human periodontal ligament cells
Wei LI ; Dong CHEN ; Kunyang LI ; Yun FAN ; Fangfang YU
Journal of Practical Stomatology 2014;(2):211-214
Objective:To study the effects of smokeless tobacco extract(ST)on the proliferation and the heat shock protein 70 (HSP70)expression of human periodontal ligament cells(hPDLCs).Methods:hPDLCs were cultured in vitro and identified by im-munohistochemistry(IHC).The cells were stimulated with ST at 0.01 6 -50 g/L respectively for 24 h,the proliferation was examine by MTT assay,HSP70 expression was detected by immunehistochemical staining and Western Blot.Results:ST inhibited the prolifer-ation and increased HSP70 expression in cytoplasm and nucleus at 0.4 -50 g/L dose dependantly.Conclusion:ST may inhibit the proliferation and increase HSP70 expression of hPDLCs in a dose depandant manner.
3.Protection of sericin to retinal microvessels and its mechanism in diabetic rats
Hongfei, YU ; Zhijun, DONG ; Tiemin, ZHANG ; Fan, YANG
Chinese Journal of Experimental Ophthalmology 2017;35(1):32-37
Background Diabetic retinopathy (DR) is a chronic inflammatory disease,with pathological changes of retinal microvessels.Studies demonstrated that sericin has anti-inflammation and anti-oxidation effects,inferring that sericin might play a protective effect on diabetic microangiopathy.Objective This study was to investigate the effects of sericin on intercellular adhesion molecule-1 (ICAM-1),vascular cell adhesion molecule-1 (VCAM-1) and Cx43 expressions in retina and explore the protection of sericin to retinal microangiopathy in diabetic rats.Methods Forty-eight specific pathogen free male SD rats were randomly divided into normal control group,diabetic model group,sericin-treated group and calcium dobesilate-treated group,with 12 rats for each group.The diabetic models were established by intraperitoneal injection of streptozotocin (STZ) for 3 consecutive days and feeding up with high lipid foods.Normal saline solution,2.4 g/(kg · d) sericin solution and 0.2 g/(kg · d) calcium dobesilate were used by gavage administration in the rats of the diabetic model group,sericin-treated group and calcium dobesilate-treated for 3 months group,respectively.The rats were sacrificed and retinal sections were prepared,and the retinal morphology was examined by hematoxylin-eosin staining.The expressions of ICAM-1,VCAM-1 and Cx43 proteins and mRNA in retinas were detected by Western blot assay and reverse transcription PCR.The use and care of the rats complied with Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission and ARVO statement.Results The retinal structure was normal in the normal control group.The swell and rupture of inter limiting membrane (ILM),scatter vascular endothelial cell nuclei breakthrough ILM and decrease of retinal ganglion cells (RGCs) were displayed in the diabetic model group;while in the sericin-treated group and calcium dobesilate-treated group,the mild thickening of ILM and disorder of retinal cells were obtained.The relative expression levels of ICAM-1 and VCAM-1 were significantly raised and those of Cx43 were reduced in the diabetic model group,sericin-treated group and calcium dobesilate-treated group when compared with the normal control group (all at P<0.05).Compared with the diabetic model group,the expressions of ICAM-1 and VCAM-1 proteins and mRNA in the sericin-treated group were significantly reduced (ICAM-1 protein:0.834 3±0.032 1 vs.0.918 9±0.042 4;VCAM-Ⅰ protein:0.726 4±0.011 2 vs.1.235 0±0.078 9;ICAM-1 mRNA:0.716 3±0.008 6 vs.0.956 8±0.012 5;VCAM-1 mRNA:0.393 7±0.035 0 vs.0.477 9±0.020 6) and those of Cx43 protein and mRNA were evidently elevated (Cx43 protein:0.133 1 ±0.015 3 vs.0.039 2±0.002 0;Cx43 mRNA:0.676 8 ±0.064 8 vs.0.430 8±0.111 3) (all at P<0.05).Conclusions Sericin can relieve retinal microangiopathy and protect retina against the pathogenesis and development of DR by down-regulating the expressions of ICAM-1,VCAM-1 and upregulating the expression of Cx43 in retinas of diabetic rats.
4.Inhibition effect of adiponectin on the levels of PC Ⅲ and HA in HSC-T6 cell
Fujun YU ; Peihong DONG ; Xufei FAN ; Yongping CHEN ; Ji LI
Journal of Chinese Physician 2011;13(2):150-153
Objective To observe the effects of adiponectin on mRNA and protein expressions of connective tissue growth factor (CTGF) in hepatic stellate cells (HSCs) and the levels of procollagen type Ⅲ (PC Ⅲ) and hyaluronic acid (HA).Methods Cultured rat HSCs were treated with different concentrations of adiponectin.CTGF mRNA and protein expressions were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western Blot.The levels of PCⅢ and HA were detected by enzymelinked immunosorbent assay (ELISA).Results Compared with the control group,the result of RT-PCR showed that the four groups had different degrees of inhibitory effect,of which group D exhibited the strongest inhibitory effect.The absorbance ratio was 1.54 ±0.18,1.21 ±0.14,0.96 ±0.10,and 0.79 ± 0.08,respectively (t =2.42,P <0.05;t =2.73,P <0.05;t =3.28,P <0.01;t =4.67,P <0.01).Western Blot also indicated that four groups had different degrees of inhibitory effect,of which D group exhibited the strongest inhibitory effect.The ratio of integral absorption was 1.54 ± 0.18,1.21 ±0.14,0.96±0.10,and 0.79 ±0.08,respectively (t =2.84,P <0.01;t =4.05,P <0.01;t =6.25,P <0.01;t =9.72,P <0.01).The levels of PCⅢ and HA secreted in culture media were also decreased.It was significantly decreased with the concentration of adiponectin increased.Conclusion Adiponectin can inhibit the levels of PC Ⅲ and HA,which may be achieved through reducing CTGF mRNA and protein expressions.
5.Imaging features of the arteriovenous malformation in maxilla
Mengda JIANG ; Xindong FAN ; Qiang YU ; Minjun DONG ; Xiaofeng TAO
Journal of Practical Radiology 2016;32(5):682-685
Objective To summarize the radiography ,CT and MRI findings of maxillary arteriovenous malformation (AVM ) . Methods Seventeen patients with maxillary AVM underwent panoramic radiography (3 patients) and enhanced MRI examination (6 patients) ,all the patient underwent enhanced CT examination .The clinical manifestation and imaging findings of maxillary AVM ,in‐cluding the location ,shape ,margin ,inner texture ,involvement of adjacent structures ,the density and signal intensity of the lesions , were analyzed .Results The lesions mainly located in molar areas (15/17) .The major clinical manifestation were repeated bleeding and acute bleeding .Other symptoms included swelling of the face ,pulsatile soft mass and anesthesia .The panoramic radiography only showed increased density .According to the change of the maxilla on enhanced CT images ,the lesions could be devided into two types :type Ⅰ ,showed intraosseous osteolytic destruction and cortical expansion (n=12);type Ⅱ ,showed“ground glass”appearance (n=5) .Bone destruction and soft tissue involvement ,widened feeding artery and draining veins ,elevated maxillary sinus were shown in all patients .External jugular vein enlargement and early enhancement (n=14) and root resorption (n= 6) were also detected . Flowing void on T1 and T2 weighted images (type Ⅰ) and hypo‐or isointense on T1WI ,hyperintense on T2WI and obvious enhance‐ment after injecting contrast material (type Ⅱ ) could observed .Conclusion Enhanced CT examination could be primarily recom‐mended and observe the extent of lesions and situation of vessels invaded of maxillary AVM .
6.Surveillance of Pathogens Distribution and Antibiotic Resistance in Lower Respiratory Tract Infection in ICU
Tianmei ZHOU ; Xiaoqin DONG ; Daojun YU ; Qiang SHEN ; Jianzhong FAN
Chinese Journal of Nosocomiology 2004;0(10):-
OBJECTIVE To investigate the distribution of pathogens and their antibiotic resistance in lower respiratory tract infection(LRTI) from intensive care unit(ICU) in our hospital,and provide basis for rational selection of clinical drugs.METHODS Pathogens were detected from qualified sputum specimens in LRTI from ICU and identified by VITEK-AMS60 automatic microbial analyzing system.Drug susceptibility was determined by KB test.RESULTS From 320 sputum specimens 367 pathogens were detected between from Jan 2007 to Mar 2008,including 261 strains(71.1%) of Gram-negative bacilli,70 strains(19.1%) of fungi,and 36 strains(9.8%) of Gram-positive cocci.21.8% Of the isolated pathogens were Acinetobacter baumannii,with 16.7% of drug-resistant rate to cefoperazone/sulbactam and over 71% to other 13 antibiotic agents.The rate of extended spectrum ?-lactamases(ESBLs) producing Escherichia coli isolates and Klebsiella pneumoniae ones were 65.2% and 72.0%,respectively,comparing to 84.6% for meticillin-resistant Staphylococcus aureus(MRSA).CONCLUSIONS Gram-negative bacilli are the major pathogens of LRTI in ICU,in which A.baumannii shows with a high rate of drug-resistance,followed by fungi,which should attract the clinician′s more attention.
7.A study on the factors influencing the prevalence of echinococciasis in Gansu Province
Dawei YU ; Pengfei GE ; Yu FENG ; Fan LI ; Yandong HOU ; Dong WANG
Chinese Journal of Endemiology 2015;34(10):746-749
Objective To investigate the factors influencing the prevalence of echinococciasis in Gansu Province.Methods With the method of stratified random sampling,all the administrative villages in pastoral areas,half pastoral areas,agriculture and urban areas in the 72 counties in Gansu Province were selected;according to the layers of the population of the county population proportion calculated,the layer investigation numbers were determined,and a total of 16 administrative villages were selected in each county.In the 16 administrative villages,more than 200 people of permanent residents were surveyed in each village (shortage was made up from a nearby village),at least 3 200 people were investigated in a county.By using the combined method of inquiry and field observation,natural factors,such as drinking water,farming and animal husbandry production data were surveyed.All data were analyzed using SPSS 19.0 statistical software.Unauy linear and multiple linear regression analysis of influencing factors and the relationship between the prevalence were analyzed.Results Unary linear regression analysis showed that longitudes,latitudes,altitudes,average annual temperature,annual rainfalls,secondary industries,animal husbandry towns,drinking water sources,the number of epidemic towns,epidemic villages,per capita net incomes in animal husbandry regions,the total number of livestock and the number of sheep were factors influencing the prevalence of echinococciasis (F =4.705,P <0.05).Latitude,altitude,animal husbandry towns,the number of epidemic towns,epidemic villages,per capita net incomes in animal husbandry regions,the total number of livestock and the number of sheep and the prevalence of echinococciasis was positively correlated (r =0.282,0.285,0.387,0.508,0.540,0.317,0.475,0.594,all P <0.05);longitude,average annual temperature,annual rainfall,secondary industries,drinking water sources and the prevalence of echinococciasis was negatively correlated (r =-0.311,-0.244,-0.244,-0.389,-0.311,all P <0.05).Multiple linear regression analysis showed that differences of interactions of annual rainfalls,drinking water sources,epidemic villages,per capita net incomes of herdsmen,secondary industries and the total number of livestock between groups were statistically significant (t =-1.822,-3.920,3.013,1.715,-1.609,3.264,all P <0.05).Conclusion The factors influencing the prevalence of echinococciasis in Gansu Province are correlated with annual rainfalls,drinking water sources,epidemic villages,per capita net incomes of herdsmen,secondary industries and the total number of livestock.
8.Development and evaluation of an inactivated bivalent vaccine against duck viral hepatitis.
Fenggui YIN ; Li JING ; Shuang ZHANG ; Meng YU ; Wanlin ZHANG ; Guobing FAN ; Xiukai DONG ; Wenjun LIU
Chinese Journal of Biotechnology 2015;31(11):1579-1588
The rapid mutation and widely spread of duck hepatitis A virus (DHAV) lead to the vast economic loss of the duck industry. To prepare and evaluate bivalent inactivated vaccine laboratory products of DHAV, 6 strains were screened from 201 DHAV-1 strains and 38 DHAV-3 strains by using serotype epidemiological analysis in most of the duck factory. Vaccine candidate strains were selected by ELD50 and LD50 tests in the 6 strains. Continuously passaged, the 5th passaged duck embryos bodies grinding fluid was selected as vaccine virus seeds. The virus seeds were treated with formaldehyde and water in oil in water (W/O/W) emulsions, making into three batches of two bivalent inactivated vaccine laboratory products. The safety test, antibody neutralization test, challenged protection and cross immune protection experiment suggested that the vaccines possessed good safety, and neutralizing antibodies were detected at 7th day and the challenged protection rate reached 90% to 100% at the 14th and 21st day. Moreover, immune duration of ducklings lasted more than five weeks. However, cross-immunity protection experiments with DHAV-SH and DHAV-FS only had 20%-30%. The two bivalent inactivated vaccine laboratory products of duck viral hepatitis were effective and reliable, providing a new method as well as a new product for DHAV prevention and control.
Animals
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Antibodies, Neutralizing
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blood
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Ducks
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virology
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Hepatitis Virus, Duck
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Hepatitis, Viral, Animal
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prevention & control
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virology
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Neutralization Tests
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Picornaviridae Infections
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prevention & control
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veterinary
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Poultry Diseases
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prevention & control
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virology
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Vaccines, Inactivated
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immunology
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Viral Hepatitis Vaccines
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immunology
9.NMR-based analysis of water soluble extracts of different Astragali Radix.
Dong TIAN ; Zhen-Yu LI ; Sheng-Ci FAN ; Jin-Ping JIA ; Xue-Mei QIN
Acta Pharmaceutica Sinica 2014;49(1):89-94
Water soluble extract (WSE) is an important index for the quality evaluation of Astragali Radix (AR). In this study, the WSE of the wild AR from Shanxi province (SX) and the cultivated AR from Gansu Province (GS) were compared. The WSEs of two types of AR were determined according to the appendix of Chinese pharmacopoeia. Then the WSEs were subjected to NMR analysis, and the obtained data were analyzed using HCA, PCA, OPLS-DA, microarray analysis, and Spearman rank analysis. In addition, the Pearson correlation of differential metabolites were also calculated. The results showed that the WSE content of GS-AR (37.80%) was higher than that of SX-AR (32.13%). The main constituent of WSE was sucrose, and other 18 compounds, including amino acids, organic acids, were also detected. Multivariate analysis revealed that SX-AR contained more choline, succinic acid, citric acid, glutamate, taurine and aspartate, while GS samples contained more sucrose, arginine and fumaric acid. In addition, the Pearson correlations between different metabolites of the two types of AR also showed apparent differences. The results suggested that the WSE of two types of AR differs not only in the content, but also in the chemical compositions. Thus, the cultivation way is important to the quality of AR. This study supplied a new method for the comparison of extract of herbal drugs.
Arginine
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analysis
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Aspartic Acid
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analysis
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Choline
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analysis
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Citric Acid
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analysis
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Drugs, Chinese Herbal
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analysis
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chemistry
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Fumarates
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analysis
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Glutamic Acid
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analysis
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Magnetic Resonance Spectroscopy
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Multivariate Analysis
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Phylogeography
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Plant Roots
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chemistry
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Plants, Medicinal
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chemistry
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Succinic Acid
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analysis
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Sucrose
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analysis
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Taurine
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analysis
10.Multi-ligaments reconstruction with single tendon with for the treatment of acute joint dislocation of the first carpometacarpal.
Yu ZHANG ; Wei-min FAN ; Nan-wei XU ; Wei-dong GU
China Journal of Orthopaedics and Traumatology 2013;26(12):1037-1040
OBJECTIVETo explore the clinical outcomes of multi-ligaments reconstruction with single tendon in treating acute joint dislocation of the first carpometacarpal.
METHODSFrom December 2008 to October 2012, 4 patients with acute dislocation of the first carpometacarpal were treated with single carpi radialis longus tendon to reconstruct periarticular four ligaments, which included dorsal ligament, palmar ligament, dorsal radiocarpal ligament, and intermetacarpal ligament between the first and second. There were 3 males and 1 female aged from 22 to 63 years old with an average of 38.7. X-ray, JAMAR grid strength testing, range of motion of carpometacarpal joint and VAS score were used to evaluate clinical outcomes.
RESULTSAll patients were followed up from 6 to 40 months with an average of 19 months. The wound were healed well at stage I. No dislocation of the first carpometacarpal joint and signs of joints degeneration occurred on X-ray at 1, 3 and 6 months after operation. JAMAR grip strength recovered from 60 percent to 90 percent of health wide. The results of ROM showed 3 cases recovered flexion and extension of joints and 1 case limited adduction. VAS score was 0 to 1.
CONCLUSIONFor acute dislocation of the first carpometacarpal joint, reconstruction four ligaments with single carpi radialis longus tendon can be considered the selected treatment,which can restores joint stability and improve joint function.
Adult ; Carpal Joints ; injuries ; surgery ; Female ; Hand Injuries ; surgery ; Humans ; Joint Dislocations ; surgery ; Ligaments, Articular ; surgery ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Tendons ; surgery ; Young Adult