Objective:The effect of Tonglin capsule was observed on type steblay interstitial nephritis of cavy.Methods: The cavy was immunized using the basement membrane of renal tubule from renal cortex of rabbit and Freund's complete adjuvant (FCA). The animal model of autoimmunity renal tubule interstitial nephrifis was made, The effect of Tonglin capsule on nephric pathological changes was observed. Results: ① The levels of creatinine (Cr) and blood uria nitrogen (BUN) in treatment groups were obviously lower than that in model group ( P

Objective:To construct an eukaryotic expression vector of enhanced green fluorescence protein(EGFP) gene driven by telomerase catalytic subunit(hTERT) gene promoter and observe the specific expression of EGFP in lung cancer cell lines.Methods:The 1100bp promoter fragment was obtained by enzyme digestion from a recombinant plasmid of pGL3-hTERTp containing the hTERT promoter.The hTERT promoter was then subcloned into the upstream of the report gene EGFP of pEGFP-1 without promoter.The expression vector pEGFP-hTERTp was successfully constructed.The vector pEGFP-N1 containing cytomegalovirus(CMV) promoter was used as a positive control.The vector pEGFP-1 without promoter was used as a negative control.The vectors were transfected into human lung cancer cell lines 95D,NCI-H446,A2,A549,LTEP-a-2,YTMLC and normal MRC-5 through lipofectamine respectively.EGFP expression was detected under the fluorescence microscope.Results:pEGFP-hTERTp was confirmed by enzyme digestion with correct result.That the EGFP expression was detected in all of eight lung cancer cells transfected with pEGFP-hTERTp,but not in MRC cells.By contrast,high intensity EGFP expression was observed in both lung tumor cells and normal cells,which were transfected with pEGFP-N1.Conclusion:The EGFP controlled by hTERT promoter can be expressed specifically in lung cancer cell lines.hTERT promoter may be used as an excellent regulation element in tumor-targeting gene therapy.

Objective:To investigate the protective effects of nuclear factor ?B inhibition on liver graft reperfusion injury during transplantation.Methods:Orthotopic liver transplantation using modified cuff technique was established in rats,animals were divided according to the grafts cold storaged in 4℃ Ringer's lactated solution with(PDTC group)or without 0.1 mol/L PDTC(control group)for 6 h.During the early stage of reperfusion,DNA binding activities of NF ?B in liver grafts were analyzed by EMSA(electrophoretic mobility shift assay),mRNA level of TNF ? and ICAM 1 by RT PCR,and activities of ALT and LDH were also detected.Results:NF ?B in liver grafts was activated at early stage of reperfusion during transplantation; PDTC treated liver displayed lower activation of NF ?B 1 h after reperfusion,whereas no difference was shown between 2 groups 6 h after reperfusion.Up regulation of TNF ? and ICAM 1 transcription,high level of ALT and LDH activities were observed in both groups during reperfusion,whereas the transcriptional up regulation and the activities of ALT and LDH in PDTC group were reduced compared with those of control group( P

Objective To evaluate the capability of 18 F-FLT uptake and investigate the early radiation response of human colorectal cancer cells HCT116 exposed to 6 MV X-rays.Methods 3.7 kBq 18F-FLT was added to HCT116 cells with different cell numbers (1.0 × 105-1.5 × 106) and cultured with different times (36,60,84 h).The 18F-FLT uptake rate was measured with a γ-counter after exposed to different does of 6 MV X-rays (0,2,4,6,8 Gy) after 24,48,and 72 h of irradiation.Then the cell uptake inhibition rate,cell proliferation,and cell cycle phase were measured.Results The uptake rate of 18F-FLT in HCT116 was (18.97 ± 1.16)％.The 18F-FLT uptake inhibition rates at 24 h after different does of irradiation (2,4,6,8 Gy) were (32.10±0.02)％,(54.46 ±0.04)％,(62.74 ±0.04)％,and (65.81 ±4.81)％,respectively,which was positively correlated with radiation dose.Conclusions The 18F-FLT uptake rate of human colorectal cancer HCT116 cells could be used to evaluate the early radiation response.

Objective To investigate the relationship between the expression of ITGAV and the radiosensitivity of NSCLC cells. Methods The expression of ITGAV in NSCLC and its relationship to the prognosis of patients who received radiotherapy were analyzed using bioinformatics methods. Differences in radiosensitivity between radio-resistant cells and parent cells were verified by clone formation experiment, and the protein expression of ITGAV was detected by Western blot. The transfection efficiency of si-ITGAV was determined by Western blot and qRT-PCR analyses. The best ITGAV interference sequence was selected to transfect A549R and H1299R cells. Clone formation experiment and flow cytometry were used to detect clone formation, apoptosis and cell cycle of A549R and H1299R cells. Results The expression of ITGAV in NSCLC tissues was significantly higher than that in normal tissues (P<0.05), and NSCLC patients with high ITGAV expression had poor prognosis. The clonogenic ability of the si-ITGAV group was significantly lower than that of the negative control group at 4, 6, 8Gy irradiation (all P<0.05). After 6 Gy irradiation, the apoptosis of the si-ITGAV group was increased (P_H1299R<0.0001, P_A549R=0.0002), the proportion of G₂/M phase cells to A549-siITGAV and H1299R-siITGAV cells was higher than that in the negative control group (P_H1299R<0.0001, P_A549R=0.0007). Conclusion Interfering with ITGAV expression can increase the radiosensitivity of NSCLC.

This study sought to clone Chinese Banna minipig inbred-line (BMI) alpha1,3-galactosyltransferase (alpha1,3-GT) gene and construct its recombinant eukaryotic expression vector. Total RNA was isolated from BMI liver. Full length cDNA of alpha1,3-GT gene was amplified by RT-PCR and cloned into pMD18-T vector to sequence. Subsequently, alpha1,3-GT gene was inserted into pEGFP-N1 to construct eukaryotic expression vector pEGFP-N1-GT. Then the reconstructed plasmid pEGFP-N1-GT was transiently transfected into human lung cancer cell line A549. The expression of alpha1,3-GT mRNA in transfected cells was detected by RT-PCR. FITC-BS-IB4 lectin was used in the direct immunofluorescence method, which was performed to observe the alpha-Gal synthesis function of BMI alpha1,3-GT in transfected cells. The results showed that full length of BMI alpha1,3-GT cDNA was 1116 bp. BMI alpha1,3-GT cDNA sequence was highly homogenous with those of mouse and bovine, and was exactly the same as the complete sequence of those of swine, pEGFP-N1-GT was confirmed by enzyme digestion and PCR. The expression of alpha1,3-GT mRNA was detected in A549 cells transfected by pEGFP-N1-GT. The expression of alpha-Gal was observed on the membrane of A549 cells transfected by pEGFP-N1-GT. Successful cloning of BMI alpha1,3-GT cDNA and construction of its eukaryotic expression vector have established a foundation for further research and application of BMI alpha1,3-GT in the fields of xenotransplantation and immunological therapy of cancer.
Animals ; Animals, Inbred Strains ; Base Sequence ; China ; Cloning, Molecular ; Galactosyltransferases ; genetics ; metabolism ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Recombinant Proteins ; genetics ; metabolism ; Sequence Analysis, DNA ; Swine ; Swine, Miniature ; genetics ; Transfection

Objective:To analyze the risk of missed diagnosis in patients with PI-RADS score>3 and negative prostate initial biopsy and to explore its risk factors.Methods:The clinical data of 268 patients with negative prostate biopsy in Northern Jiangsu People's Hospital from May 2013 to December 2018 were retrospectively analyzed. The patients were divided into observation group (PI-RADS score>3) and control group (PI-RADS score≤ 3) according to different PI-RADS scores. There were insignificant differences in age [(67.4(60.0, 74.0)years and 65.6(66.5, 72.0)years], prostate volume of initial biopsy [62.4(40.0, 72.0)ml and 60.8(38.0, 77.0)ml], biopsy cores [ 20.6(18.0, 22.0)cores and 20.4(18.0, 22.0)cores] between the observation group (n=124) and the control group(n=144)(all P>0.05). But there were significant differences in PSA [17.5(6.5, 23.0)ng/ml and 11.5(6.3, 12.0)ng/ml], PSAD[0.316(0.128, 0.363)ng/ml 2 and 0.211(0.106, 0.256)ng/ml 2], prostate inflammation of the initial biopsy [70 (56.5%) and 32 (22.2%)] between the observation group and the control group(all P<0.05). According to the follow-up results after the initial biopsy, the two groups of repeated biopsy were compared.Furthermore, Logistic regression was used to conduct univariate and multivariate analysis to explore the risk factors of patients with PI-RADS>3 for positive repeated biopsy. At the same time, the receiver operating characteristic curve (ROC curve) was used to analyze the accuracy of the risk factors. Results:There were significant differences in repeated biopsy rate [ 27.4%(34/124)and 14.6%(21/144)], CsPCa detection rate[ 41.4%(14/34) and 4.8%(1/21)]between the observation group and the control group(all P<0.05). The positive rate of repeated biopsy in the observation group (41.1%) was higher than that in the control group (23.8%), but there was no statistical difference ( P=0.248). The risk of positive repeated biopsies in the observation group was 2.24 times than that in the control group. Univariate analysis found repeated biopsy PSA ( P =0.02, OR=1.438, 95% CI 1.161-1.896), PSA ratio (repeated biopsy PSA/initial biopsy PSA) ( P=0.011, OR=10.087, 95% CI 1.714-59.36) were risk factors for positive of repeated biopsy in patients with PI-RADS score >3. Multivariate analysis also found that repeated biopsy PSA ( P=0.017, OR=1.15, 95% CI 1.076-2.123), PSA ratio ( P=0.032, OR=10.2, 95% CI 0.883-116.168) were risk factors for positive repeated biopsy. ROC curve analysis, the accuracy of repeated biopsy PSA (AUC=0.971, P<0.001, 95% CI 0.926-1.000), PSA ratio (AUC=0.839, P=0.001, 95% CI0.707-0.971) to predict positive of repeated biopsy were high. The cut-off values were 21.3 ng/ml and 1.4, respectively. The accuracy was higher when combines repeated biopsy PSA with PSA ratio (AUC=0.993, P<0.001, 95% CI 0.974-1.000). Conclusions:Patients with negative PI-RADS score > 3 have a higher risk of missed diagnosis of CsPCa than those with PI-RADS score≤3. When PSA>21.3 ng/ml and PSA ratio>1.4 during follow-up, the possibility of missed diagnosis in the initial biopsy is high.

Objective:To study the clinical characteristics of ureaplasma urealyticum (UU) infection in preterm infants with gestational age <34 weeks.Methods:From January 2017 to December 2021, premature infants with gestational age <34 weeks admitted to neonatal department of our hospital were enrolled in this prospective cohort study. UU-DNA from respiratory tract samples were examined using quantitative fluorescence polymerase chain reaction method. The infants were assigned into UU (+) group and UU (-) group. Perinatal factors and clinical characteristics were compared between the two groups.Results:A total of 182 preterm infants were enrolled, including 59 cases (32.4%) in UU (+) group and 123 (67.6%) in UU (-) group. UU (+) group had significantly lower gestational age and birth weight and significantly higher incidences of vaginal delivery, premature rupture of membranes (PROM) >18 h and maternal chorioamnionitis than UU (-) group ( P<0.05). Compared with UU (-) group, UU (+) group had significantly higher leucocyte count, neutrophil count and interleukin-6 at 1, 24 and 72 h after birth ( P<0.05). No significant differences existed in C-reactive protein and procalcitonin between the two groups at each time point ( P>0.05). In UU (+) group, the incidences of intrauterine pulmonary infection and bronchopulmonary dysplasia (BPD) were higher and the incidence of respiratory distress syndrome was lower than UU (-) group ( P<0.05). No significant differences existed in the incidences of intraventricular hemorrhage, periventricular leukomalacia, feeding intolerance, necrotizing enterocolitis, retinopathy of prematurity between the two groups ( P>0.0 5). UU (+) group had significantly longer duration of oxygen therapy than UU (-) group ( P<0.05). No significant differences existed in the duration of invasive mechanical ventilation and hospital stay between the two groups ( P>0.05). Conclusions:Preterm infants <34 weeks with positive UU in respiratory tract secretions have higher incidences of vaginal delivery, PROM>18 h and maternal chorioamnionitis. Leukocyte and neutrophil count and interleukin -6 are higher in these infants. They need prolonged oxygen therapy and have increased risks of intrauterine pulmonary infection and BPD.

Objective:To explore the feasibility and safety of the clinical application of the diagnosis and treatment mode combining rapid frozen pathological examination of prostate biopsy tissue with radical prostatectomy.Methods:Suspected prostate cancer patients with PSA>10 ng/ml and PI-RADS score ≥4 in, Northern Jiangsu People's Hospital from April to September 2021 were collected. The included patients underwent mpMRI/TRUS image fusion-guided transperineal prostate targeted biopsy with 16G biopsy needle, 2-3 needles for biopsy, and rapid frozen pathological examination. Robot-assisted laparoscopic radical prostatectomy (RALP) was performed immediately for patients with prostate cancer with rapid freezing pathology. For undiagnosed prostate cancer, 18G biopsy needle for prostate targeted + systematic biopsy were used, 18-22 needles for systematic biopsy, and routine pathological examination. The baseline data, frozen pathological results, perioperative conditions, pathological results and follow-up data of all patients were collected.Results:Eleven patients were included in the study, the mean age of the patients was 69.9(66-73) years, the mean BMI was 22.8(19-26) kg/m 2, the mean PSA was 23.2(14.25-32.00), the mean prostate volume was 45(32-52) ml, mean PSAD 0.54(0.33-0.75). PI-RADS score was 4 in 3 cases and 5 in 8 cases; digital rectal examination was positive in 5 cases. All 11 cases underwent rapid freezing and the pathological results showed that: 9 cases were prostate adenocarcinoma, and RALP was performed immediately. The operation time was 111.5(96-126) min, the intraoperative blood loss was 78.9(55-105) ml, and the postoperative extubation time was 4.3(3.5-5.0) days, postoperative hospital stay 5.8(5.0-6.5) days. Postoperative pathology showed that Gleason score 3+ 4=7 in 1 case, 4+ 3=7 in 3 cases, 8 points in 4 cases, and 10 points in 1 case; 3 cases had positive resection margins, and 1 case had seminal vesicle invasion, the average number of dissected lymph nodes was 10.9 (8.5-14.0), and there was no tumor metastasis. Pathological T staging included 2 cases of T 2b stage, 5 cases of T 2c stage, 1 case of T 3a stage, and 1 case of T 3b stage. Two patients were undiagnosed by rapid freezing pathology, of which one was prostate adenocarcinoma with a Gleason score of 4+ 3=7, and then received RALP; the other one was prostate inflammation. 11 patients were followed up; the postoperative follow-up time was 3-7 months, with an average of 5.2 months. Among the 10 patients who underwent RALP, 8 patients recovered urinary continence 2 weeks after surgery, and all recovered within 2 months after surgery. Three patients with positive surgical margins were given regular androgen deprivation therapy in the second week after surgery. PSA did not drop below 0.1 ng/ml in patients with positive margins and seminal vesicle invasion 3 months after surgery. No complications of Clavien grade Ⅰ or higher occurred after operation and during follow-up. Conclusions:For patients with high suspicion of prostate cancer, rapid frozen pathological examination of prostate biopsy tissue is performed. RALP is performed immediately for patients with prostate cancer. The results show that this diagnosis and treatment model could be safe and feasible.