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AIM: To establish a ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to determination the plasma concentration of clozapine and compare the bioequivalence of a generic clozapine tablet with Clozaril
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Objective:To explore the effect and mechanism of microRNA (miRNA)-4320 on the proliferation and migration of gastric cancer MGC803 cells.Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-4320 in four gastric cancer cell lines(MGC803, HS-746T, SGC7901, BGC823) MGC803 cells were infected with recombinant lentivirus carrying miR-4320 interference fragments or blank lentivirus, and set as si-miR-4320 group and NC group. Thiazole blue colorimetry and Transwell small box experiment were used to detect the proliferation and migration of MGC803 cells after miR-4320 was down-regulated. The bioinformatics software RNAhybrid was used to predict the target gene of miR-4320. The targeting relationship between miR-4320 and target gene was verified by dual-luciferase reporter gene experiment. qRT-PCR and Western blot were used to detect the expression of miR-4320 target gene. Measurement data were expressed as mean ± standard deviation ( ± s), and t-test or one-way ANOVA was used for comparison between groups. Results:The expression of miR-4320 in the four gastric cancer cell lines was significantly higher than that of normal gastric mucosal epithelial cells ( P<0.01). The expression of miR-4320 in MGC803 cells in the NC group and the si-miR-4320 group were 8.19±1.00 and 1.09±0.31, respectively. The miR-4320 interference fragment significantly reduced the expression of miR-4320 ( P<0.01). The absorbance of MGC803 cells in the si-miR-4320 group was significantly lower than that of the NC group ( P<0.05), and the migration ability was significantly lower than that of the NC group ( P<0.01). Suppressor of cytokine signaling1 ( SOCSI) is the target gene of miR-4320. Compared with the NC group, the SOCS1 gene expression in the si-miR-4320 group was significantly up-regulated ( P<0.01). Conclusions:The expression of miR-4320 is increased in gastric cancer cell lines. Down-regulating the expression of miR-4320 can inhibit the proliferation and migration of gastric cancer MGC803 cells by inducing the expression of SOCS1 gene.
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Objective:To explore the effect of down-regulation of long non-coding RNA (lncRNA) CTB-191K22.5 on the proliferation and invasion of colorectal cancer SW480 cells and the molecular mechanism.Methods:The TCGA database was used to analyze the expression differences of CTB-191K22.5 in colorectal cancer tissues and normal tissues. The CTB-191K22.5 inhibitor (Anti-CTB-191K22.5) and negative inhibitor (Control) were transfected into colorectal cancer SW480 cells, denoted as Observation group and Control group, real-time quantitative polymerase chain reaction (qRT) -PCR) was used to evaluate the inhibitory effect. MTT method and Transwell chamber method were used to evaluate the proliferation and invasion of SW480 cells. Western blot was used to evaluate the protein levels of PI 3K/AKT/mTOR signaling pathway in SW480 cells. The bioinformatics software starBase v2.0 was used to predict the target genes of CTB-191K22.5. qRT-PCR was used to evaluate the expression of CTB-191K22.5 target gene in SW480 cells. Measurement data were expressed as Mean±SD, and t-test was used for comparison between two groups. Results:Compared with normal tissues, the expression of CTB-191K22.5 in colorectal cancer tissues was significantly increased ( P<0.01). The expression of CTB-191K22.5 in SW480 cells of the Control group and Observation group were 6.60±0.85 and 1.08±0.21, respectively. The expression level of CTB-191K22.5 decreased after transfection with Anti-CTB-191K22.5 ( P<0.01). Compared with the Control group, the SW480 cell proliferation ability of the Observation group decreased ( P<0.01). The invasion numbers of SW480 cells in the Control group and Observation group were (135.4 ± 16.29) and (42.24±14.59), respectively. The invasion ability of SW480 cells decreased after transfection with Anti-CTB-191K22.5 ( P<0.01). Compared with the Control group, the expression levels of PI 3K/AKT/mTOR signaling pathway protein in SW480 cells in the Observation group decreased. miR-326 may be the target gene of CTB-191K22.5. Compared with the Control group, transfection with Anti-CTB-191K22.5 significantly increased the expression level of miR-326 in SW480 cells ( P<0.01). Conclusion:CTB-191K22.5 is highly expressed in colorectal cancer tissues, and down-regulation of CTB-191K22.5 may inhibit the proliferation and invasion of colorectal cancer SW480 cells by targeting miR-326.
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Objective:To investigate the effect of long non-coding RNA (lncRNA) HAGLR on the proliferation and migration of gastric cancer cells by inhibiting the expression of microRNA (miRNA, miR)-93-5p.Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of HAGLR in gastric cancer cell lines (HS-746T, BGC823, SGC7901, MGC803) and normal gastric mucosal epithelial cells (GES-1). Selected the cell line with the lowest HAGLR expression and transfected with the negative control plasmid (negative control group) or HAGLR-high-expression plasmid (HAGLR group) respectively. The MTS method and the scratch healing test were used to detect the proliferation and migration ability of the cells after transfection. The bioinformatics software miRcode database was used to predict the target gene of HAGLR, and the dual luciferase reporter gene experiment was used to verify the binding of HAGLR to the target gene. qRT-PCR was used to detect the expression of the target gene. Western blot was used to detect the expression of Hippo signaling pathway. The software SPSS 21.0 was used to conduct statistical analysis. The t test was used for comparison between two groups, and the one-way analysis of variance was used for comparison between multiple groups. Results:Compared with GES-1 cells, the expression level of HAGLR in gastric cancer cell lines was lower (all P<0.05), and the cell line with the lowest HAGLR expression was SGC7901 cells ( P<0.01). The HAGLR expression in SGC7901 cells in the HAGLR group and the negative control group were 1.03±0.13 and 9.75±1.10, respectively. The expression level of HAGLR in the negative control group was significantly lower than that in the HAGLR group ( t=7.87, P<0.01). Compared with the negative control group, the absorbance of SGC7901 cells in the HAGLR group was significantly reduced ( P<0.05), and the scratch healing rate was significantly reduced ( P<0.01). The miRcode database showd that HAGLR and miR-93-5p have complementary binding sites. The dual luciferase reporter gene experiment showed that HAGLR can complement miR-93-5p ( P<0.01). Compared with the negative control group, the expression of miR-93-5p in SGC7901 cells in the HAGLR group was significantly reduced ( P<0.01), and the expression of Hippo signaling pathway protein was significantly reduced (all P<0.01). Conclusions:HAGLR is low expressed in gastric cancer cell lines. HAGLR inhibits the proliferation and migration of gastric cancer SGC7901 cells by negatively regulating miR-93-5p.
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Objective:To explore the effect of microRNA (miRNA)-3126-5p on the proliferation and migration of colorectal cancer cells by inhibiting the expression of LIM and SH3 protein 1 ( LASP1). Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-3126-5p in colorectal cancer cell lines (HT-29, HCT116, LoVo, SW480) and normal intestinal mucosal epithelial cells (HIEC). The cell line with the lowest expression level was selected as the experimental object. The experiment was divided into 2 groups: the negative control group (transfected with miR-NC) and the miR-3126-5p group (transfected with miR-3126-5p). Cells of each group were collected 48h after transfection. qRT-PCR method was used to detect the expression level of miR-3126-5p in each group. The MTS method and the scratch healing experiment were used to detect the proliferation level and migration ability of the cells in each group. The bioinformatics software microRNA.org and the dual-luciferase reporter gene experiment were used to predict and verify the target genes of miR-3126-5p, respectively. qRT-PCR and Western blot were used to detect the expression levels of target genes in each group of cells. Measurement data were expressed as mean±standard deviation ( Mean± SD), t test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. Results:Compared with normal intestinal mucosal epithelial cells (HIEC), the expression level of colorectal cancer cell line miR-3126-5p was significantly reduced ( P<0.05), and the cell line with the lowest expression level was HCT116 cells ( P<0.01). The expression of miR-3126-5p in HCT116 cells in the negative control group and miR-3126-5p group were (1.05±0.16) and (7.91±1.26) respectively, and the difference was statistically significant ( t=5.40, P<0.01). Compared with the negative control group, the proliferation ability of HCT116 cells in the miR-3126-5p group was significantly reduced ( t=4.52, P<0.05), and the migration ability was significantly reduced ( P<0.01). microRNA.org shows that miR-3126-5p has complementary binding sites with LIM and SH3 protein 1 ( LASP1) gene mRNA. miR-3126-5p can target LASP1 mRNA ( P<0.01). Compared with the negative control group, the expression of LASP1 gene in HCT116 cells of the miR-3126-5p group was significantly reduced ( t=4.56, P<0.01). Conclusion:The expression of miR-3126-5p in colorectal cancer cell lines is low, and miR-3126-5p can reduce the proliferation and migration ability of colorectal cancer HCT116 cells by inhibiting the expression of the target gene LASP1.
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Objective To explore the clinical effect of live Bacillus Subtilis and Enterococcus Faecium granulescombined with blue light irradiation in the treatment of neonatal jaundice.Methods 90 patients with neonatal jaundice were randomly divided into control group and observation group.The control group received the blue light treatment,the observation group was given blue light irradiation combined with live Bacillus Subtilis and Enterococcus Faecium granules treatment,the patients were treated for 7 days.The clinical efficacy,serum bilirubin and main indicators of complex constant time were observed in the two groups.Results After treatment,the total effective rate of the observation group was 95.6％,which of the control group was 68.9％,the difference was statistically significant (x2 =5.473,P ＜ O.05).After treatment for 1 day,3 days,7 days,the serum bilirubin values of the observation group were (185.6 ± 25.1) μmol/L,(136.7 ± 19.1) μmol/L,(82.1 ± 10.3) μmol/L,respectively,which of the control group were (205.3 ± 26.4) μmol/L,(184.1 ± 20.3) μmol/L,(128.4 ± 16.7) μmol/L,the differences between the two groups were statistically significant (t =3.628,11.408,15.829,all P ＜ 0.05).The recovery time of serum bilirubin in the observation group was (7.1 ± 2.4) d,which of the control group was (12.9 ±3.1) d,there was significant difference between the two groups (t =9.924,P ＜ 0.05).The hospitalization time of observation group was (3.5 ± 1.1) d,which of the control group was (5.8 ± 1.5) d,the difference between the two groups was statistically significant (t =8.295,P ＜ 0.05).Conclusion Live Bacillus Subtilis and Enterococcus Faecium granules combined with blue light irradiation in the treatment of neonatal jaundice is helpful to improve the clinical efficacy,reduce the treatment time and improve the clinical symptoms.
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Objective To investigate the curative effect of cephalosporins in the treatment of neonatal infectious pneumonia and the effect on intestinal microflora.MethodsA total of 124 cases of neonatal pneumonia in our hospital were divided into the cephalosporin group (40cases), the piperacillin group (38 cases) and the combined treatment group (46 cases), and 40 healthy neonates were selected as healthy group.The clinical efficacy was compared.The intestinal bacterial genus of the four groups was examined on the 5th day after treatment, including Enterobacter, Bacteroides, Enterococcus, Bifidobacterium and Lactobacillus.ResultsThe cure rates of the cephalosporin group, the piperacillin group and the combined group were 82.5%, 81.57% and 89.13%, respectively.The healing time of the above three groups was (5.3±0.2) d, (5.5±0.3) d and (5.2±0.3) d, respectively.Enterobacteriaceae, Bacteroides, Enterococcus and Lactobacillus were significantly more in the above three groups than the healthy group, and Bifidobacterium was fewer than healthy group (P<0.05).There was no significant difference between the cephalosporin group and piperacillin group.ConclusionThe curative effect of cephalosporins is similar to other antibiotics in the treatment of neonatal infectious pneumonia.The former can effectively relieve alteration of intestinal flora, combined with other drugs.Irrational use of antibiotics woll increase alteration of intestinal flora.
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Objective To explore the diagnostic value of serum procalcitonin(PCT) and C-reactive protein (CRP) in using of antibiotics in patients with acute bronchial asthma attack.Methods Sixty cases with acute bronchial asthma attack were divided into allergen-induced group (18 cases),viral infectioninduced group (14 cases) and bacteria infection-induced group (28 cases).The serum PCT level was examined by immune luminous method.The CRP level was examined by enzyme linked immune scattered method.Results The serum PCT and CRP level was (25.7 ± ll.2)μg/L and (50.3 ± 17.5) mg/L in bacteria infection-induced group,which was significantly higher than that in viral infection-induced group and allergen-induced group [(0.7 ± 0.2) μ g/L,(6.1 ± 0.3) mg/L and (0.2 ± 0.1) μ g/L,(3.5 ± 0.4) mg/L,P ＜ 0.05].There was no significant difference in the serum PCT and CRP level between viral infectioninduced group and allergen-induced group (P ＞ 0.05).There was positive correlation between PCT and CRP in bacterial infection group (r =0.932,P ＜ 0.05),while no correlation existed between PCT and CRP in viral infection-induced group and allergen-induced group (r =-0.021,-0.103,P＞0.05).CRP≥10 mg/L andPCT ≥0.5 μ g/L was as positive criteria.The positive rate of serum CRP and PCT in bacteria infection-induced group was 89.3％(25/28) and 92.9％(26/28),which was higher than that in viral infection-induced group and allergen-induced group [28.6％ (4/14),64.3％ (9/14) and 22.2％ (4/18),33.3％ (6/18),P ＜0.05],and the positive rate of serum CRP and PCT in viral infection-induced group was higher than that in allergen-induced group,there was significant difference (P ＜ 0.05).Conclusion The serum PCT and CRP level can correctly identify early bacteria infection patients with acute bronchial asthma attack.
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Objective: To observe the clinical efficacy of combined electroacupuncture and nerve growth factor (NGF) injection at acupoints in the treatment of common fibular nerve paralysis and provide evidences for integrative Chinese & western medicine against diabetic peripheral neuropathy (DPN). Methods: Forty subjects were randomized into two groups and NGF injection; and control group was given herbal suffocation, oral Dibazol and compound vitamin B and Mecobalamin Injection. The clinical symptoms and nerve conduction velocity were observed and compared. Results: The cure rate was higher in treatment group than in control group (P<0.05); after treatment, the nerve conduction velocity was improved in both groups (P<0.01), with a significant improvement in treatment group than in control group (P<0.01). Conclusion: Combined electro-acupuncture and NGF injection at acupoints is quite effective in the treatment of common fibular nerve paralysis.
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Objective To investigate the changes of PPARr mRNA and protein expressions in rats with ANP. Methods 36 rats were randomly divided into sham group and ANP group. The rats were sacrificed 3 h, 6 h,12 h after ANP induction, the levels of serum amylase were measured, the pancreatic pathological changes were determined and the expressions of pancreatic PPARr mRNA and protein was examined by RT- PCR and immunohistochemistry. Results In ANP group, the level of serum amylase at 6h was (7170.83± 1635.59) U/L, the scores of pathological changes were 6.67±1.03 and 13.00±2.36, which were much higher than those of sham group (P<0.01) ; the PPARr mRNA expression was 0.18±0.05, and there were no obvious differences compared with that of sham group (0.22±0.03 ) ; PPARr protein expression was 4.17 ±0.98, which was significantly higher than that of sham group (1.83±0.71, P<0.05). Conclusions Inflammatory injury resulted in increased deactivation of pancreatic acinar PPARr, meanwhile PPARr gene expression was inhibited by feedback.