Objective To construct the lentiviral RNA interference vector targeting TPX2 and to obtain the human cervical cancer HeLa cell strain stably infected by TPX2-shRNA for studying the relationship between human cervical carcinoma and TPX2 gene.Methods By targeting TPX2 gene,four double-stranded DNA hairpin structures corresponding to shRNA were designed,synthesized and connected with Pglv2-U6-Puro to construct the recombinant plasmids.Then these recombinant plasmids were transformed into DH5α competent cells.The positive clone was extracted and transfected into 293T cells for virus packages after sequenced correctly.Human cervical carcinoma HeLa cell infected by these recombinant lentiviral was screened by Puromycin,then stable cell strain was obtained.The silencing effect of TPX2 in HeLa cell was detected by RT-fluorescent quantitative PCR and Western blot.Cell cycle and cell apoptosis wer detected by Flow cytometry.Results Sequencing results confirmed that 5 lentiviral was packaged successfully.The steady cell strain transfered TPX2-shRNA was screened with 0.4 μg/mL puromycin.HeLa cells infected by recombinant lentivirus all play the gene silencing effect especially in the group of TPX2-shRNA-1.In the group of TPX2-shRNA-1,TPX2mRNA (0.21 ± 0.07) and protein (0.19 ± 0.28) rela tive expression level is lower than those in the control group (1.08±0.07) (P＜0.01) and(0.64±0.03) (P＜ 0.01)respectively;G2 and S-phase cells are higher than those in the control group (P＜0.05)and the apoptosis rate was significantly more than those in the control group (P＜0.05).Conclusions The effective TPX2 genetic interference sequence was obtained,lentiviral vectors carrying TPX2shRNA was successfully constructed,and the HeLa cell strain with TPX2 silenced was successfully screened,which lay the research foundation for the study of the role of TPX2 in cervical cancer.

Objective:To investigate the correlation between polymorphisms of serine hydroxymethyltransferase1 gene and the adverse reactions of high-dose methotrexate (HD-MTX) in children with acute lymphoblastic leukemia (ALL). Methods:A total of 51 patients with ALL were treated with HD-MTX, and clinical manifestations after HD-MTX treatment were evaluated retrospectively. cD-NA was obtained from mRNA. The polymorphisms of SHMT1 gene containing rs1979277, rs3783, rs1979276, and rs12952556 sites were tested by denaturing gradient gel electrophoresis and direct sequencing. Effects of SHMT1 gene polymorphisms on HD-MTX ad-verse reactions were evaluated. Results:Severe adverse reactions in ALL patients treated with HD-MTX appeared to be mainly neutro-penia and hepatoadverse reactions. The frequency distributions of rs3783 (C>G), rs1979276 (C>T), rs12952556 (A>G), and rs1979277 (C>T) were the same. The polymorphisms of rs1979277 showed no correlation with neutropenia (P>0.05) but rs1979277 CT and TT genotypes were correlated with hepatoadverse reactions (CT: OR=0.129, 95% CI: 0.020 to 0.817, P=0.03; TT: OR=0.103, 95% CI:0.017 to 0.620, P=0.013). Conclusion: No correlation was found between the combination of rs1979277, rs3783, rs1979276, rs12952556, and neutropenia, but one or more of these loci may reduce the risk of hepatoadverse reactions.

Through retrospective analysis of the clinical and laboratory data of 1 466 inpatients with type 2 diabetes mellitus(T2DM),we investigated the prevalence of chronic kidney disease (CKD) and analyzed the risk factors.The prevalence of CKD in hospitalized patients with T2DM was 52.25％.In the patients with CKD,protein urine was present in 93.47％ of the cases,27.93％ of them had glomerular filtration rate(eGFR) ≤60 ml · min-1 · 1.73 m-2,damage of renal tubular function was present in 24.28％,and abnormal renal imaging in 14.88％.Logistic regression showed that age,body mass index(BMI),duration of diabetes,systolic blood pressure,serum uric acid,low density lipoprotein-cholesterol (LDL-C),and smoking were independently associated with patients of T2 DM and CKD.The prevalence of CKD was increased with aging,diabetic course,BMI,and LDL-C.CKD is a common chronic complication in patients with T2DM,especially in patients with prolonged course,advanced age,and obesity.Much attention should be paid to early detection of CKD in patients with diabetes.In addition to detecting urinary protein and eGFR,renal tubular function and morphological examination should also be included.

To evaluate the neuroprotective effect of nerve growth factor ( NGF ) on acute angle-closure glaucoma patients after trabeculetomy.METHODS: Patients with viral keratitis who underwent trabeculectomy in treatment of acute angle - closure glaucoma in Xiamen Eye Center of Xiamen University from December 2011 to October 2013 were selected and completed the treatment, 61 eyes of 45 cases were followed up. The treatment group of 23 cases (32 eyes) with acute angle-closure glaucoma patients were treated by NGF gel for 3mo after trabeculetomy, while in the control group 22 cases (29 eyes) were treated by normal saline replaced NGF. All patients were followed up for at least 12mo, visual acuity, perimetry, retinal nerve fiber layer ( RNFL) thickness and cup/disc ratio of the patients were followed up during the treatment. The safety of topical use of NGF gel was also evaluated.RESULTS:ln all patients, the intraocular pressure ( lOP) was successfully controlled under 21mmHg and the visual acuity was markedly increased in the affected eye after trabeculectomy. Compared with control group, the postoperative lOP and visual acuity had no significant differences in treatment group (P>0. 05). The average light sensitivity of perimetry and mean defect were better than that in control group postoperative 6 and 12mo; The results of optical coherent tomography ( OCT ) and Heiderburg Retina Tomography ( HRT )-Ⅲ showed that RNFL thickness was significantly greater than that in control group, while cup/disc ratio significantly less than that in control group postoperative 6 and 12mo. Ocular surface damage, corneal endothelium to reduce and other eye complication were no observed in treatment group.CONCLUSlON:Acute angle-closure glaucoma treated by NGF gel after operation is effective and safe.

Objective:To compare the effectiveness of fire needle versus Western medicine in the treatment of herpes zoster. Methods:Randomized controlled trials comparing fire needle with Western medicine in the treatment of herpes zoster were identified using 8 databases. A meta-analysis was performed using RevMan 5.3 software. Results:Eight trials involving 569 patients were included in this meta-analysis, and the results showed that fire needle was superior to Western medicine comparing the effective rate [risk ratio (RR)=1.13, 95％ confidence interval (CI): 1.06 to 1.20;P=0.0002], the visual analog scale (VAS) score [mean difference(MD)=–7.95, 95％ CI: –10.71 to –5.20;P<0.00001], time of pain disappearance (MD=–7.61, 95％CI: –9.38 to –5.84;P<0.00001), time of blister-stop (MD=–1.34, 95％CI: –1.51 to –1.18;P<0.00001), time of crusted scab (MD=–2.92, 95％CI: –3.62 to –2.23;P<0.00001), and time of scab off (MD=–4.64, 95％CI: –5.83 to –3.46;P<0.00001). In addition, a significantly lower incidence of postherpetic neuralgia was found in the fire needle group in 30 d (RR=0.23, 95％CI: 0.11 to 0.51;P=0.0002) and 60 d (RR=0.33, 95％CI: 0.12 to 0.91; P=0.03) after treatment. Conclusion:Fire needle has a favorable effect in increasing the effective rate, relieving pain, recovering skin lesions and decreasing incidence of postherpetic neuralgia in the treatment of herpes zoster. However, considering the limitations in this study, the findings should be interpreted cautiously.

PURPOSE: We tried to find out whether the nadir PSA level after hormone therapy affected the progression into hormone-refractory prostate cancer (HRPC). MATERIALS AND METHODS: We reviewed the progressive status and the survival of the 177 patients with stage C or D prostate cancer who had received hormone therapies. The relative efficacy of the nadir PSA level for predicting the progression into HRPC was evaluated by the receiver operating characteristic (ROC) analysis. RESULTS: 85.4% of patients responded to the treatment and 78% of responders progressed into HRPC. Median time to nadir PSA level after hormone therapy and to HRPC were 8.1 and 24.0 months, respectively. The nadir PSA levels were under 0.2ng/ml in 31%, 0.2-1.0ng/ml in 23%, 1.1-10ng/ml in 42%, and over 10ng/ml in 5% of the responders (n=151). As the nadir PSA levels were lower, pretreatment PSA levels, Gleason score and the number of cases progressing into HRPC were significantly lower (p<0.05). In addition, the nadir PSA level was inversely correlated with the interval to the establishment of HRPC (r= 0.465, p<0.05). By univariate analysis, the bone metastasis, the nadir PSA level, PSA level at six months after treatment and pretreatment PSA level were associated with the progression into HRPC. Only the nadir PSA level was an independent factor by multivariate analysis. ROC analysis disclosed an accuracy of 86.2% for the nadir PSA level to predict the progression into HRPC after two years. By setting the lower limit of the nadir PSA level to 1.1ng/ml, the sensitivity was 80.3% and the specificity was 83.8%, being most adequate. CONCLUSIONS: The nadir PSA level after hormone therapy may be the most important factor that can predict the progression into HRPC. Also, in consideration of sensitivity and specificity, it would be adequate to set the lower limit of the nadir PSA level to 1.1ng/ml.
Humans ; Multivariate Analysis ; Neoplasm Grading ; Neoplasm Metastasis ; Prognosis ; Prostate* ; Prostate-Specific Antigen* ; Prostatic Neoplasms* ; ROC Curve ; Sensitivity and Specificity

No abstract available.
Animals ; Nitric Oxide Synthase* ; Nitric Oxide* ; Rats* ; Spermatic Cord Torsion* ; Testis*

Objective To investigate the association of genetic polymorphisms of intercellular adhesion molecule 1 (ICAM-1) with diabetic peripheral neuropathy (DPN). Methods A total of 607 type 2 diabetes patients from the Affiliated Hospital of Weifang Medical University were enrolled in this study between June 2013 and December 2014. Rs5498 (A/G K469E) and rs1799969 (G/A R241G) in the ICAM-1 gene were genotyped by using TaqMan allelic discrimination in 295 patients with DPN and 312 subjects without DPN. The distribution of these two SNPs and the genetic influence of ICAM-1 gene polymorphisms on the development of DPN were conducted. Results Genotype distributions of both SNPs were coincided with Hardy-Weinberg equilibrium in the two groups. SNP rs1799969 (G/A R241G) in the ICAM-1 gene showed a high GG genotypic frequency at 96.8%(non DPN) and 99.0%(DPN) respectively. SNP rs5498 (A/G K469E) represented AA and AG genotypes. The values were AA 48.7%/AG 39.4%in non DPN group and AA 51.5%/AG 41.7%in DPN group. There were no significant differences in genotypic distributions and allele frequencies of SNPs rs1799969 (G/A R241G) and rs5498 (A/G K469E) between the patients with DPN group and patients without DPN group (P>0.05). The dominant(AA+AG)/GG and additive (GG/AA) models of rs5498 (A/G K469E) were associated with higher risk of DPN (ORadjusted=1.585, 1.575 respectively, P<0.05). To carry A allele was related to the susceptibility of DPN. There was no such association in genetic models of rs1799969 (G/A R241G) and DPN pathogenesis. Conclusion The present study provides evidence that SNP rs5498 E469K (A/G) in the ICAM-1 gene is associated with susceptibility of DPN, and the carrying A allele appears to be a risk of DPN.

Objective To explore the value of the plasma miR-193a-5p level on diagnosis and monitoring the response to treatment in acute myeloid leukemia (AML). Methods Peripheral blood samples were obtained from AML patients enrolled in hematology department of the Second Hospital of Shanxi Medical University from July 2015 to December 2015, including 30 de novo AML patients, 9 patients in complete remission (CR) and 6 patients in relapse. Peripheral blood samples from 15 healthy people were randomly choosed as the health control group. Plasma miR-193a-5p expression levels were detected by using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Results The plasma miR-193a-5p relative expression level of AML patients group [0.465 6 (0.103 1-5.000 2)] was obviously lower than that of health control group [0.766 1 (0.052 1-3.134 4)] (U= 122, P= 0.018 7). The plasma miR-193a-5p relative expression levels of de novo group and relapse AML group were significantly lower than those of CR group and health control group (P<0.05), and there was no significant difference between the CR group and health control group (U= 56, P= 0.511 9). No significant correlation was found between the plasma miR-193a-5p level and age, gender, blast percentage of the bone marrow, peripheral blood leukocyte count, platelet count, CD34+cells'percentage and so on. Conclusion The decreased plasma miR-193a-5p expression level can be served as a new and noninvasive biomarker for the evaluation of diagnosis and treatment in AML.

ObjectiveTo investigate the effect of the Yap1 gene and tanshinone ⅡA on the proliferation, migration, and invasion abilities of Huh-7 hepatoma cells. MethodsA total of 10 pairs of human hepatocellular carcinoma (HCC) samples and adjacent tissue samples were collected in Zhongnan Hospital of Wuhan University from June 1 to December 1, 2019. Quantitative real-time PCR and Western blotting were used to measure the expression of the Yap1 gene and phenotype-related molecules. MTT cell proliferation detection reagent was used to measure the inhibition rate of cell proliferation after the treatment with different concentrations of tanshinone ⅡA. Western blotting was used to measure the changes in the expression of apoptosis-and migration-related markers after different interventions. Flow cytometry and Transwell assay were used to measure apoptosis and cell migration and invasion abilities. The data of 375 cases of liver cancer and 50 cases of relatively normal liver tissue samples were downloaded from The Cancer Genome Atlas, including clinicopathological information. The t-test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. ResultsIn 8 of the 10 pairs of HCC samples and adjacent tissue samples, HCC samples had significantly higher expression of Yap1 than the adjacent tissue samples. Compared with the normal human liver epithelial cells L02, the Huh-7 and HCCL-M3 hepatoma cells had a significant increase in the expression of Yap1. The silencing efficiency of si-Yap1-3 transfection reached 87.004% at the protein level. MTT results showed that tanshinone ⅡA effectively inhibited the proliferation of Huh-7 cells, with a half inhibitory concentration of 8.683 μmol/L. After the cells were treated with si-Yap1-3 and tanshinone ⅡA, there was an increase in the expression of the downstream marker for proliferation and migration E-cadherin and a reduction in the expression of vimentin, and the results of Transwell assay showed that compared with the si-NC group, the tanshinone ⅡA+si-Yap1-3 group had significant reductions in the migration and invasion abilities of Huh-7 cells (migration: 43.19±2.88 vs 132.20±10.03, t=8.527, P=0.001; invasion: 53.95±4.20 vs 179.10±11.11, t=4.484, P=0.011). The group treated with si-Yap1-3 and tanshinone ⅡA had an increase in the expression of the apoptosis-related marker Bax and a reduction in the expression of Bcl-2, as well as a significantly higher early apoptosis rate than the si-NC group (2598% vs 9.21%, χ2=4.078, P＜0.05). ConclusionOncogene Yap1 silencing combined with tanshinone ⅡA can promote the apoptosis of Huh-7 hepatoma cells and inhibit their migration and invasion, which can provide certain guiding significance for clinical medication.