Objective To study the influencing factors of digital image white balance of the biological microscope and the way to obtain optimized image.Methods Color temperature meter was used to measure the influences of the factors and their correlations, and SASS software was employed for statistical analysis.Results Object lens might increase the color temperature by 200 to 300 K, LBD color filter could enhance it by 400 to 1 800 K. Reducing field stop by 90% might decrease color temperature by 400 K. ND6 filter, ND25 filter and aperture diaphragm had no influences on color temperature.Conclusion The electric and optical parameters of biological microscope have to be adjusted to gain proper white balance for digital image.

Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Arteriovenous Malformations ; complications ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Magnetic Resonance Imaging ; Middle Aged ; Multiple Sclerosis ; Spinal Cord Diseases ; complications ; metabolism ; pathology

Objective To investigate the relationship between Klotho and autophagy in sepsisinduced acute kidney injury mice model.Methods The male healthy Balb/c mice were used to establish the model of sepsis-induced acute kidney injury by using cecal ligation and puncture (CLP).Mice were sacrificed at 3 h,6 h,12 h,1 d,2 d,3 d,and 5 d after CLP (n =12 for each interval) and on 1 d 6 mice in sham group as well as 6 mice in normal group were sacrificed at the same time.Scr and BUN in the blood serum were detected.The HE and PAS staining were employed for observation on the histopathological changes in kidney tissues under light microscope.The autophagosomes were observed under transmission electron microscope (TEM).The renal protein of Klotho,LC3 and P62 were detected by using Western blot and Immunohistochemistry.Statistical analyses were performed using Student's t-test by SPSS 23.0.software.Results Scr and BUN increased significantly after CLP,especially on 1 d,respectively (165.64 ± 20.56) μmol/L and (45.51 ± 4.05) mmol/L.HE and PAS staining showed renal tissue was damaged obviously 1 d after CLP,as indicated by desquamation of the brush border of proximal tubular epithelial cells,appearance of bare basement membrane,and interstitial inflammatory cell infiltration.Under TEM,autophagosomes and phagocytosis were observed.Compared with sham group,the expression of Klotho protein decreased gradually from 3 h to 1 d and dropped to the trough at 1 d (t =51.851,P ＜0.01),then resumed gradually from 2 d to 5 d.On the contrary,the activation of autophagy increased as indicated by the expression of LC3-Ⅱ/L3-Ⅰ and p62.Autophagy was induced gradually from 3 h to 1 d and reached peak at 1 d,then declined gradually from 2 d to 5 d (P ＜ 0.01).The protein of Klotho and LC3-Ⅱ mainly distributed in renal tubular cytoplasm,and Klotho was reduced significantly (t =-8.371,P ＜ 0.01) and LC3-Ⅱ appeared in high density remarkably (t =4.995,P =0.001) on 1 d after CLP.Conclusions Klotho protein reduction and autophagy protein increase were observed in sepsis-induced acute kidney injury,and the expressions of Klotho and autophagy acted out in certain extent of time dependence.

Objective:To prepare and preliminarily identify the monoclonal antibodies(mAbs) specifically against 3D protein of Enterovirus 71(EV71),using bioinformatics to predict the epitopes of 3D,with HBc protein as a carrier.Methods: Artificial screening of 3D protein epitope sequences by bioinformatic method,inserted into the major immunodominant region(MIR) area of Hepatitis B virus core protein(HBc),to construct the recombinant protein.BALB/c mice were immunized with the recombinant virus like particles(VLPs),to prepare the mAbs against 3D protein of EV71.Affinity chromatography technology was used to purify the mAb.The indirect ELISA,ELISPOT,immunofluorescence and immunohistochemistry staining methods were used to identify the characteristic of the mAb.Results: We displayed 3D(aa34-43),3D(aa61-76) and 3D(aa151-164) epitopes by constructing fusion protein using HBc VLPs as a vector,after hybridization,one positive hybridoma cell line(3E1) was selected by ELISA.The isotype of 3E1 was IgG2a.The results of immunofluorescence and immunohistochemistry staining assay showed that the mAb 3E1 could specifically recognize EV71.Conclusion: The prepared mAb 3E1 can specifically recognizes the EV71,which laid the foundation for the detection of virus and further study on 3D protein,and verified the bioinformatics technology combined with HBc carrier displaying peptides could prepare mAb quickly and efficiently.

OBJECTIVETo investigate the effect of bone mesenchymal stem cell (BMSC) transplantation on repair of glomerular podocytes and on the Nephrin expression in rats with puromycin aminonucleoside (PAN) -induced nephrosis.

METHODSForty-five Sprague-Dawley rats were randomly divided into three groups (n=15 each): a nephrosis model group that received a single intraperitoneal injection of PAN (0.15 mg/g); a BMSC transplantation group that received a single intraperitoneal injection of PAN (0.15 mg/g) followed by BMSC transfusion; a control group that received a single intraperitoneal injection of normal saline. Ten days after injection, the rats were sacrificed. The 24 hrs urinary protein content and serum albumin and cholesterol levels were measured 24 hrs before sacrifice. Changes of glomerular podocytes were observed under an electron microscope. Brdu labeled positive cells in kidneys were measured by immunohistochemical technology. RT-PCR and Western blot were used to assess the expression of mRNA and protein of Nephrin.

RESULTSIn the nephrosis model group, urinary protein and blood cholesterol contents increased, plasma albumin content decreased compared with those in the control group. Extensive fusion of podocyte foot processes was observed in the nephrosis model group. The BMSC transplantation group had decreased urinary protein and blood cholesterol contents and increased plasma albumin content compared with the nephrosis model group. Fusion of podocyte foot processes was also improved. Brdu labeled positive cells were seen in kidneys in the BMSC transplantation group, but not in the nephrosis model and the control groups. Nephrin mRNA and protein expression decreased significantly in the nephrosis model group compared with that in the control group. The BMSC transplantation group had increased Nephrin mRNA and protein expression compared with the nephrosis model group.

CONCLUSIONSBMSCs can repair glomerular podocytes in PAN-induced nephrosis rats, and the changes of Nephrin expression may be involved in the process.

Animals ; Bromodeoxyuridine ; metabolism ; Kidney ; pathology ; Male ; Membrane Proteins ; genetics ; Mesenchymal Stem Cell Transplantation ; Nephrosis, Lipoid ; chemically induced ; pathology ; therapy ; Podocytes ; pathology ; Puromycin Aminonucleoside ; toxicity ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

Objective: To analyze the effect of acupuncture versus hormone replacement therapy (HRT) for premature ovarian insufficiency (POI). Methods: China National Knowledge Infrastructure (CNKI), Wanfang Academic Journal Full-text Database (Wanfang), Chongqing VIP Database (CQVIP), China Biology Medicine Disc (CBM), Web of Science, Cochrane Library, PubMed, and Excerpta Medica Database (EMBASE) were searched up to January 31st, 2019 to identify randomized controlled trials (RCTs) evaluating the effect of acupuncture for POI. The primary outcome was the level of basal serum follicle- stimulating hormone (FSH). Secondary outcomes included serum levels of luteinizing hormone (LH), estradiol (E2) and anti-Müllerian hormone (AMH). Two authors extracted data independently and assessed the risk of bias and the methodological quality using the Cochrane's tool. Meta-analysis was conducted by RevMan version 5.3. Results: Eight eligible RCTs with a total of 496 POI patients were included in the meta-analysis. The pooled results showed that there was a significant reduction in the basal serum FSH level (MD=-5.82, 95％CI:-9.76 to -1.87, I2=82％, P=0.004) and a remarkable elevation in the basal E2 level (SMD=0.93, 95％CI: 0.34 to 1.52, I2=88％, P=0.002) in the acupuncture group when compared with the control. Subgroup analysis showed that compared with HRT, a significant decrease in the FSH level was observed in both acupuncture alone (MD=-4.53, 95％CI:-8.96 to -0.10, I2=73％, P=0.04) and acupuncture plus HRT (MD=-9.60, 95％CI:-17.60 to -1.61, I2=50％, P=0.02), while a remarkable elevation of E2 was only found in acupuncture plus HRT (SMD=1.43, 95％CI: 1.03 to 1.82, I2=0％, P<0.00001). There was no significant difference in the LH level between acupuncture and HRT (MD=-3.16, 95％CI:-9.41 to 3.10, I2=0％, P=0.32), only one trial reported AMH, and no significant difference was found between acupuncture and HRT. Conclusion: The present study indicated that acupuncture had an advantage over HRT in reducing serum FSH level and increasing serum E2 level in women with POI. However, evidence supporting the finding is limited due to the small sample size, potential methodological flaws and significant heterogeneity. Hence, this conclusion still needs to be verified by high-quality RCTs.

OBJECTIVETo investigate the distribution of burn pathogens and their antibiotic resistance in a burn unit, so as to provide reference for clinical practice.

METHODSThree hundred and forty-eight burn patients hospitalized in our department were enrolled in this study. The pathogens isolated from the wounds, blood, venous catheter, sputum, urine, purulent discharge of wounds in these patients, and their antibiotic resistance were surveyed by retrospective analysis from Jan, 2001 to Dec, 2006.

RESULTSTotal-ly 464 strains were isolated, among which Gram negative (G-) bacilli accounted for 52.6%, Gram positive microorganisms (G+) accounted for 40.5%, and fungi accounted for 6.9%. The main pathogens were Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter species and Escherichia coli, among which Staphylococcus aureus (MRSA) was predominant (93.5%). MRSA was 100% resistant to levofloxacin, penicillium, oxacillin, and it was also resistant to other antibiotics except Vancomycin. The resistance rate of Pseudomonas aeruginosa to Cefoperazone/Sulbactam, Imipenem and cefepime were 15.8%, 36.8%, 33.3%, respectively.

CONCLUSIONStaphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter species and Escherichia coli were predominant in the burn unit,among them Staphylococcus aureus and Acinetobacter were more resistant to antibiotics.

Acinetobacter baumannii ; drug effects ; isolation & purification ; Burn Units ; Burns ; microbiology ; Cross Infection ; microbiology ; Drug Resistance, Bacterial ; Escherichia coli ; drug effects ; isolation & purification ; Humans ; Pseudomonas aeruginosa ; drug effects ; isolation & purification ; Retrospective Studies ; Staphylococcus aureus ; drug effects ; isolation & purification

The purpose of the present study is to establish a rapid and effective PCR method for the identification of B. multicinctus. Based on sequence alignment of B. multicinctus and its adulterants, we found that Cyt b gene is a good molecular genetic marker for the authentication of B. multicinctus. On the basis of the sequence data, a pair of highly specialized primers was designed. The templates were extracted by the DNA purification system. Key factors such as annealing temperature, concentration of Taq enzyme and cycle numbers were analyzed and optimized. The modified PCR program consisted of an initial denaturation step at 95 degrees C for 5 min, followed by 30 cycles of 95 degrees C for 30 s and 55 degrees C for 45 s and a final extension at 72 degrees C for 5 min. Thirteen samples of B. multicinctus were identified accurately from their 20 adulterants in 4 hours. The results indicated it is a highly accurate, rapid and applicable method for the authentication of B. multicinctus.
Animals ; Bungarus ; classification ; genetics ; Cytochromes b ; genetics ; DNA Primers ; genetics ; Drug Contamination ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Sequence Alignment ; Sequence Analysis, DNA

OBJECTIVETo investigate the effect of farnesoid-X-receptor (FXR) antagonist Z-guggulsterone in an in vivo high-fat fed apolipoprotein E knockout (ApoE(-/-)) mice model of myocardial ischemia/reperfusion (I/R).

METHODSMale ApoE(-/-) mice were randomly divided into three groups: standard ApoE(-/-) group (fed with standard mouse diet for 12 weeks before myocardial I/R procedure, n = 18), high-fat ApoE(-/-) group (fed with high-fat mouse diet for 12 weeks before myocardial I/R procedure, n = 22), and high-fat ApoE(-/-) + FXR antagonist group(fed with high-fat mouse diet for 12 weeks and received FXR antagonist Z-Guggulsterone 30 minutes before myocardial I/R procedure, n = 17). The expression of FXR was detected by real-time quantitative-PCR. Myocardial infarct size was determined by Evans blue/TTC double staining methods. Myocardial apoptosis was determined by in situ TUNEL technique. Markers of the mitochondrial-mediated apoptotic pathway (cytochrome c release, caspase-9 activity, and BAX and BCL-2 levels), endoplasmic reticulum stress apoptotic pathway (caspase-12 activity and CHOP level), and death receptor apoptotic pathway (caspase-8 activity, and Fas and FasL levels) were also measured.

RESULTFXR expression (3.7-fold higher, P < 0.01), myocardial infarct size [(62.1 ± 7.0)% vs. (33.8 ± 5.8)%, P < 0.01] and myocardial apoptosis index[ (36.8 ± 5.7)% vs. (17.2 ± 3.8)%, P < 0.01]were all significantly higher in high-fat ApoE(-/-) group than those in standard ApoE(-/-) group. Compared with high-fat ApoE(-/-) group, myocardial infarct size [(24.4 ± 4.7)% vs. (62.1 ± 7.0)%, P < 0.01] and myocardial apoptosis index [(13.8 ± 2.7)% vs. (36.8 ± 5.7)%, P < 0.01] were significantly reduced in high-fat ApoE(-/-) + FXR antagonist group. Moreover, levels of mitochondrial-mediated apoptotic pathway markers (cytochrome c release, caspase-9 activity, and BAX/BCL-2 levels) and endoplasmic reticulum stress apoptotic pathway markers (caspase-12 activity and CHOP level) were significantly lower in high-fat ApoE(-/-) + FXR antagonist group than those in high-fat ApoE(-/-) group (all P < 0.01). Levels of death receptor apoptotic pathway markers (caspase-8 activity, and Fas and FasL levels) were similar between high-fat ApoE(-/-) group and high-fat ApoE(-/-) + FXR antagonist group.

CONCLUSIONFXR antagonist alleviates myocardial reperfusion injury in cholesterol-fed ApoE(-/-) mice via inhibition of the mitochondrial-mediated and endoplasmic-reticulum stress pathway.

Animals ; Apolipoproteins E ; genetics ; Apoptosis ; drug effects ; Caspase 9 ; metabolism ; Cholesterol, Dietary ; administration & dosage ; Cytochromes c ; metabolism ; Disease Models, Animal ; Endoplasmic Reticulum Stress ; Male ; Mice ; Mice, Knockout ; Myocardial Reperfusion Injury ; metabolism ; pathology ; prevention & control ; Pregnenediones ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Receptors, Cytoplasmic and Nuclear ; antagonists & inhibitors ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo investigate co-expression of CD99/MIC2 and anaplastic lymphoma kinase (ALK) protein in anaplastic large-cell lymphoma (ALCL) tissues and Karpas 299 cells and its significance.

METHODSClinical prognoses and ALK protein expressions of 25 cases of ALCL were reviewed retrospectively, the median duration of survival was analyzed for patients with ALK(+) ALCL and ALK(-) ALCL. Histological and immunohistochemical staining were applied to other 25 cases of ALCL and paraffin-embedded tissue from human anaplastic large-cell lymphoma Karpas 299 cells to detect the protein of CD99 and ALK.

RESULTSOf former 25 cases of ALCL, median duration of survival for ALK(+) patients was 59 months, whereas 20 months for ALK(-) patients. The prognosis of ALK(+) group was better than that of ALK(-) group, survival curves of these two groups showed statistically significant (P < 0.05). CD99 was positive in 18 cases (72.0%) while negative in 7 cases (28.0%) of the latter 25 ALCL, ALK was positive in 19 cases (76.0%) while negative in 6 cases (24.0%); Of 19 ALK(+) ALCL, 16 (84.2%) cases co-expressed CD99-ALK; and in 6 ALK(-) ALCL, 2(33.3%) were CD99-ALK double negative, the expression of CD99 protein strongly correlated with that of ALK protein (P < 0.05). ALK and CD99 protein expressed in Karpas 299 cells with diffuse distribution.

CONCLUSIONSCD99 highly expressed in ALCL, and showed high rate of co-expression with ALK. CD99 protein expression could be considered as a helpful diagnostic and prognostic factor of ALCL, especially for ALK(+) ALCL.

12E7 Antigen ; Adolescent ; Adult ; Aged ; Antigens, CD ; metabolism ; Cell Adhesion Molecules ; metabolism ; Cell Line, Tumor ; Female ; Humans ; Lymphoma, Large-Cell, Anaplastic ; diagnosis ; metabolism ; Male ; Middle Aged ; Prognosis ; Receptor Protein-Tyrosine Kinases ; metabolism ; Retrospective Studies ; Young Adult