Objective:To determine the effects of silencing glucose regulated protein ( GRP94 ) on the proliferation and apoptosis of breast carcinoma MCF7 cells. Methods:Chemically synthesized siRNA targeting GRP94 gene and transfected into MCF7 cells used by Liopfectamine RNAIMAX. The mRNA and protein expression levels of GRP94,cyclinD1,Bax and Bcl-2 were detected by Real-time PCR and Western blot. CCK8 assay was used to detect the effect of specific GRP94 siRNA on cell proliferation and the effect on cell cycle and apoptosis were analyzed by flow cytometry and Hoechst 33258 staining. Results:Compared with the siRNA-NC cells, the expression of GRP94 was significantly down-regulated in MCF7 cells. Knockdown of GRP94 in MCF7 cells decreased cell proliferation and promoted cell apoptosis. The expression of cyclinD1and Bcl-2levels were significantly reduced, and Bax level was increased in siRNA-GRP94 MCF7 cells. Conclusion: The siRNA-mediated GRP94 silence significantly inhibits MCF7 cell proliferation,promoted cell apoptosis by down-regulating cyclin D1,Bcl-2 expression and up-regulating the Bax expression in MCF7 cells.

Objective To investigate the expressions of TLR2 and TLR4 in patients with psoriasis vulgaris and the efrect of methotrexate(MTX)on them.so as to explore the therapeutic mechanism of MTX in psoriasis vulgafis.Methods Forty-three patients with psoriasis vulgaris were recruited into the study together with 30 normal human controls.Oral MTX was given to patients with an interval of 12 hours for three times per week until the control of conditions followed by 4 weeks of mainmining treatment.The dosage of MTX was 5 mg initially and decreased to 2.5 mg in the maintaining period.Flow cytometry was used to detect the expression of TLR2 and TLR4 in peripheral blood CD14~+ cells from the controls and patients at baseline,4 and 8 weeks after the beginning of treatment.Results The expression rate of TLR2 and TLR4 in CD14~+ cells was(92.6±4.3)%and(48.5±4.6)%,respectively,in untreated patients,significantly higher than that in normal controls(botll P＜0.01).A significant increase was observed in the expression rate of TLR2 and TLR4 in patients with active psoriasis compared with those with inactive psoriasis [(97.5±4.1)%vs(87.6±5.6)%,(55.3±5.8)%vs(40.7±7.1)%,both P＜0.05].Eigh weeks after the beginning of treatment with MTX.the expression rate of TLR2 and TLR4 significantly decreased to (79.6±6.7)%and(34.6±5.9)%.respectively(both P＜0.05).The psoriasis area and severity index(PASI)score had no significant correlation with the expression rate of TLR2 or TLR4(r=0.24.0.27,both P＞0.05).Conclusions TLR2,TLR4 and innate immune response mediated by both receptors play an important role in the pathogenesis of psoriasis.MTX may exert its therapeutic effect on psoriasis by inhibiting the expression of TLR2 and TLR4.

Aim To investigate ALEX1 gene expres-sion in cervical cancer tissues and adjacent non-can-cerous tissues, and to explore the ALEX1 genetic influ-ence on cell proliferation,cycle and apoptosis of human cervical cancer cell line HeLa. Methods ALEX1 protein expression in cervical cancers and in non-can-cerous cervical tissues was evaluated using immunohis-tochemical method. A small interference RNA targeting ALEX1 gene was transfected into HeLa cells′, and the effect of ALEX1 interference on HeLa cells′ cycle and apoptosis was analysed by flow cytometry. The effect of ALEX1 interference on HeLa cells′ proliferation and sensitivity to resveratrol was analysed by CCK-8 assay. Results ALEX1 protein expression was significantly increased in cervical cancer tissues compared with non-cancerous tissues. HeLa cells′proliferation was inhibi-ted compared with control group and blank group. He-La cells′ sensitivity to resveratrol was enhanced com-pared with control group blank group. Conclution SiRNA silencing of ALEX1 gene could significantly in-hibit HeLa cells′ proliferation and enhance resveratrol ability of inhibiting HeLa cells′proliferation.

OBJECTIVE:To establish a method of microbial limit test for liquid preparation of Iron sucrose injection before filtration and sterilization.METHODS:According to the microbial limit test in the 1005 and general rules 1006 of 2015 edition of Chinese Pharmacopoeia (vol.Ⅴ),plate method and membrane filtration method were used to measure total number of aerobic bacteria (Staphylococcus aureus,Pseudomonas aeruginosa,Bacillus subtilis,Candida albicans,Aspergillus niger) and total number of molds and yeasts (C.albicans,A.niger).The optimal test method was obtained by comparing the bacterial recoveries.RESULTS:By plate method,the recoveries of P aeruginosa and B.subtilis were 2％ and 5％.The test sample was diluted 10 times with pH 7.0 sodium chloride-peptone buffer solution,and the bacterial recoveries were in the range of 88％ to 96％;but he medium was dark in color.By membrane filtration method,without rinse solution,the bacterial recoveries in the range of 88％ to 95％.Add rinse solution,the bacterial recoveries were in the range of 91％ to 103％.After validated,the recoveries of menbrane filtration method with tlushing fluid ranged 50％-200 ％,which was in line with the requnements.CONCLUSIONS:The membrane filtration method established in this experiment has higher bacterial recovery rate than the plate method.The bacterial recoveries rate were higher after adding rinse solution,and no dark substance in the surface of filter membrane affect the accotmt.It can be used as the microbial limit test method for preparation liquid of Iron sucrose injection before filtration and sterilization.

Objective:To investigate the effects of ALEX1 overexpression on cell proliferation and apoptosis of human breast cancer cell line MCF-7.Methods: MCF-7 cells were infected recombinant lentivirus LV5-ALEX1 and the negative control lentivirus LV5-NC,respectively.After 72 h, the expression of ALEX1 was detected by Real-Time PCR and Western blot.CCK8 assay were performed to observe the proliferation ability after 24 h, 48 h, 72 h, 96 h.The effect of overexpression ALEX1 on apoptosis was determined by flow cytometry.The level of Bax,BCL-2 and active caspase3 was detected by Western blot.Results:The mRNA level of ALEX1 markedly increased after 72 h(165.81±12.14 vs 52.29±2.32,P<0.01).In CCK8 assay,the results revealed that the cell pro-liferation was inhibited compared with control group in 48 h,72 h,96 h( P<0.05).The results revealed that overexpression of ALEX1 enhanced MCF-7 apoptosis(20.55%±2.50 % vs 3.60%±1.614%,P<0.05).The results by Western blot showed that the protein levels of Bax and active caspase were increased in LV5-ALEX1 group compared with LV5-NC group.However,the protein levels of BCL-2 was decreased in LV5-ALEX1 group compared with LV5-NC group.Conclusion:Overexpression of ALEX1 significantly reduced MCF-7 cancer cells proliferation and induced MCF-7 cells apoptosis.

OBJECTIVETo diagnose chromosomal abnormalities in amniotic fluid cells by combining karyotyping and single nucleotide polymorphism array (SNP-array) analysis, and to explore the application of SNP-array in routine clinical practice.

METHODSConventional G banding was used to karyotype a fetal amniotic fluid sample and the corresponding peripheral blood samples from the parents, followed by SNP-array analysis of the fetal genomic DNA from the amniotic fluid.

RESULTSThe karyotype of the amniocytes was 47, XX, +mar. The marker chromosome was further identified as psu idic (22) (q11.2) by SNP-array analysis, revealing tetraploidy of a 1.7 Mb fragment in 22q11.1-q11.2 interval that involves the critical region for Cat eye syndrome.

CONCLUSIONA rare chromosomal abnormality was identified by combining conventional G banding and SNP-array. The high resolution SNP-array could provide more detailed information for determining the origin of chromosomal abnormalities.

Adult ; Amniotic Fluid ; cytology ; Aneuploidy ; Chromosome Disorders ; genetics ; Chromosomes, Human, Pair 22 ; genetics ; Eye Abnormalities ; genetics ; Female ; Humans ; Isochromosomes ; Karyotyping ; Polymorphism, Single Nucleotide ; Pregnancy ; Tetraploidy

Objective:To study the binding characteristics of horse-derived P[12] rotavirus GST-VP8*-Horse P[12]E403 protein to oligosaccharides and saliva receptors, provides an important scientific basis for the cross-species transmission and the mechanism of interaction between the bodies.Methods:The E. coli expression system was used to express and purify the horse-derived P[12] rotavirus GST-VP8*-Horse P[12]E403 protein. The receptor binding characteristics of this genotype were analyzed by saliva and oligosaccharide binding experiments. Results:Horse-derived GST-VP8*-Horse P[12]E403 protein binds well with mucin core 2 sugar, but does not bind to other oligosaccharides such as A, B, Lewis, and HBGAs in saliva.Conclusions:The potential receptor of VP8*-Horse P[12]E403 protein may be mucin core 2, and it did not bind to human saliva.

BACKGROUND: Patients with mass-forming pancreatitis (MFP) or pancreatic ductal adenocarcinoma (PDAC) presented similar clinical symptoms, but required different treatment approaches and had different survival outcomes. This meta-analysis aimed to compare the diagnostic performance of contrast-enhanced ultrasound (CEUS) and contrast-enhanced computed tomography (CECT) in differentiating MFP from PDAC. METHODS: A literature search was performed in the PubMed, EMBASE (Ovid), Cochrane Library (CENTRAL), China National Knowledge Infrastructure (CNKI), Weipu (VIP), and WanFang databases to identify original studies published from inception to August 20, 2021. Studies reporting the diagnostic performances of CEUS and CECT for differentiating MFP from PDAC were included. The meta-analysis was performed with Stata 15.0 software. The outcomes included the pooled sensitivity, specificity, positive likelihood ratio (+LR), negative likelihood ratio (-LR), diagnostic odds ratio (DOR), and summary receiver operating characteristic (SROC) curves of CEUS and CECT. Meta-regression was conducted to investigate heterogeneity. Bayesian network meta-analysis was conducted to indirectly compare the overall diagnostic performance. RESULTS: Twenty-six studies with 2115 pancreatic masses were included. The pooled sensitivity and specificity of CEUS for MFP were 82% (95% confidence interval [CI], 73%-88%; I2  = 0.00%) and 95% (95% CI, 90%-97%; I2  = 63.44%), respectively; the overall +LR, -LR, and DOR values were 15.12 (95% CI, 7.61-30.01), 0.19 (95% CI, 0.13-0.29), and 78.91 (95% CI, 30.94-201.27), respectively; and the area under the SROC curve (AUC) was 0.90 (95% CI, 0.87-92). However, the overall sensitivity and specificity of CECT were 81% (95% CI, 75-85%; I2  = 66.37%) and 94% (95% CI, 90-96%; I2  = 74.87%); the overall +LR, -LR, and DOR values were 12.91 (95% CI, 7.86-21.20), 0.21 (95% CI, 0.16-0.27), and 62.53 (95% CI, 34.45-113.51), respectively; and, the SROC AUC was 0.92 (95% CI, 0.90-0.94). The overall diagnostic accuracy of CEUS was comparable to that of CECT for the differential diagnosis of MFP and PDAC (relative DOR 1.26, 95% CI [0.42-3.83], P  > 0.05). CONCLUSIONS CEUS and CECT have comparable diagnostic performance for differentiating MFP from PDAC, and should be considered as mutually complementary diagnostic tools for suspected focal pancreatic lesions.
Humans ; Contrast Media ; Bayes Theorem ; Tomography, X-Ray Computed/methods* ; Pancreatic Neoplasms/diagnostic imaging* ; Carcinoma, Pancreatic Ductal/diagnostic imaging* ; Sensitivity and Specificity ; Pancreatitis/diagnostic imaging* ; Ultrasonography/methods*