Effects of probiotics on intestinal flora, intestinal function and T lymphocyte level in patients with cervical cancer after radiotherapy / 中国基层医药

Objective:To investigate the effects of probiotics on intestinal flora, intestinal function, and T lymphocyte level in patients with cervical cancer after radiotherapy.Methods:A total of 92 patients with cervical cancer who underwent pelvic radiotherapy in The Second Affiliated Hospital of Zhengzhou University from September 2020 to February 2022 were included in this study. They were randomly divided into control and experimental groups ( n = 46/group). The patients in the experimental group took probiotics during radiotherapy, while the patients in the control group did not take probiotics during radiotherapy. The amount of intestinal flora, D-lactic acid, diamine oxidase, and T lymphocyte subset levels pre- and post-radiotherapy were compared between the two groups. Urinary lactulose (L) and mannitol (M) concentrations were determined in each group. Urinary excretion ratios of L to M were calculated. Results:After 10, 15, and 20 times of radiotherapy and after all radiotherapies, the amount of Escherichia coli and Enterococcus in the experimental group was significantly lower than that in the control group ( F = 128.60, 224.99, all P < 0.05). The amount of Bifidobacteria and Lactobacilli in the experimental group was significantly higher than that in the control group ( F = 2 065.46, 948.23, both P < 0.05). After 10, 15, and 20 times of radiotherapy and after all radiotherapies, plasma D-lactic acid level in the experimental group was (9.34 ± 1.63) μg/L, (9.15 ± 1.36) μg/L, (8.68 ± 1.06) μg/L, and (8.05 ± 0.82) μg/L, respectively. After 10, 15, and 20 times of radiotherapy and after all radiotherapies, plasma diamine oxidase level in the experimental group was (86.34 ± 20.25) μg/L, (84.28 ± 17.45) μg/L, (80.40 ± 13.35) μg/L, and (76.85 ± 10.87) μg/L, respectively, and urinary excretion ratio of L to M in the experimental group was (1.84 ± 0.16), (1.55 ± 0.12), (1.26 ± 0.09), (0.98 ± 0.06), respectively, all of which were significantly lower than those in the control group ( F = 121.60, 31.73, 417.84, all P < 0.05). After 10, 15, and 20 times of radiotherapy and after all radiotherapies, CD4 + level in the experimental group was (39.80 ± 4.90)%, (40.92 ± 5.30)%, (42.52 ± 6.14)%, (43.83 ± 6.55)%, respectively, CD4 +/CD8 + was (1.52 ± 0.25), (1.63 ± 0.22), (1.71 ± 0.39), (1.83 ± 0.22), respectively, all of which were significantly higher than those in the control group ( F = 58.69, 31.07, all P < 0.05). Conclusion:Probiotics can improve the status of intestinal flora and intestinal barrier function in patients with cervical cancer after radiotherapy, and simultaneously improve the cellular immune function of patients.

Silencing lncRNA UCA1 affects radiosensitivity of glioma cells by up-regulating miR-873-5p expression / 中华放射肿瘤学杂志

Objective:To investigate the effect of lncRNA UCA1 on the radiosensitivity of in vitro cultured glioma cell lines SHG-44, U87 and U251 by regulating the miR-873-5p expression. Methods:The survival of glioma cells SHG-44, U87 and U251 treated with different radiation intensities (0, 2, 4, 6 and 8 Gy) was detected by colony formation assay. The expression levels of UCA1 in glioma cells SHG-44, U87 and U251 were measured by qRT-PCR. The radiation-resistant glioma cells U87 and U251 were selected for subsequent study. After silencing UCA1 expression and/or over-expressing miR-873-5p, the cell survival rate was detected by colony formation assay, and the cell apoptosis rate was determined by flow cytometry. The dual luciferase reporter gene assay and qRT-PCR were employed to verify the targeting relationship between UCA1 and miR-873-5p.Results:UCA1 was up-regulated in the radiation-resistant U87 and U251 cells. Silencing UCA1 or over-expressing miR-873-5p inhibited the survival of U87 and U251 cells, and promoted the cell apoptosis induced by radiation exposure. miR-873-5p was a target gene of UCA1, and UCA1 negatively regulated the expression of miR-873-5p. The inhibition of miR-873-5p could reverse the effect of silencing UCA1 on the radiosensitivity of glioma cells. Silencing UCA1 increased the inhibitory effect of radiation on the glioma cell U251 xenografts.Conclusion:Silencing UCA1 inhibits the survival of glioma cells and promotes the cell apoptosis by up-regulating the expression of miR-873-5p, thereby increasing the radiosensitivity of glioma cells.

Radiosensitivity of lncrna linc00909 targeting mir-548-3p on colorectal cancer cells / 中华放射肿瘤学杂志

Objective:To investigate whether lncRNA LINC00909 affected the radiosensitivity of colorectal cancer cells by targeting miR-548-3p.Methods:The expression levels of LINC00909 and miR-584-3p in colorectal cancer and adjacent tissues were detected by qRT-PCR. The colorectal cancer cells SW480 and SW620 were cultured in vitro, and transfected with si-NC, si-LINC00909, miR-NC, miR-584-3p mimics, si-LINC00909, and anti-miR-NC and si-LINC00909, and anti-miR-584-3p, respectively. The cells were irradiated with a dose of 4 Gy. The cell survival fraction and sensitization enhancement ratio (SER) were detected by clone formation assay. Cell proliferation was detected by MTT assay. Cell migration and invasion were assessed by Trans well chamber assay. The targeting relationship between LINC00909 and miR-584-3p was confirmed by dual luciferase reporter assay. The effect of interfering with the expression of LINC00909 or inhibiting the expression of miR-584-3p on the weight of the xenograft tumor after irradiation was evaluated by subcutaneous xenograft experiment in nude mice. Results:The expression level of LINC00909 in colorectal cancer tissues was significantly up-regulated ( P<0.05), whereas the expression level of miR-584-3p was significantly down-regulated ( P<0.05). After interfering with the expression of LINC00909 or miR-584-3p overexpression, the cell survival fraction score was significantly reduced ( P<0.05), the SERs were 2.017 and 1.762, and cell proliferation, migration and invasion were suppressed (all P<0.05). Dual luciferase reporter assay confirmed that LINC00909 could target and bind to miR-584-3p. After interfering with the expression of LINC00909, the weight of the transplanted tumor was significantly reduced ( P<0.05), whereas the weight of the transplanted tumor was significantly increased after co-transfection with anti-miR-584-3p ( P<0.05). Conclusion:Interfering with the expression of LINC00909 can inhibit the proliferation, migration and invasion ability of colorectal cancer cells by up-regulating the expression of miR-548-3p, thereby enhancing the cell radiosensitivity.

LncRNA CCAT1 enhances radiosensitivity of human pancreatic cancer cells PANC-1 by targeting miR-130b-3p / 中华放射医学与防护杂志

Objective To investigate the effect of lncRNA CCAT1 and miR-130b-3p on the radiosensitivity of human pancreatic cancer cells PANC-1.Methods Real-time PCR was used to detect the relative expression levels of CCAT1 and miR-130b-3p in pancreatic cancer tissues and cell lines including PANC-1 cells irradiated with 2 Gy X-rays.After silencing CCAT1 and/or inhibiting miR-130b-3p expression,cell apoptosis rate,Caspase 3 activity and cell survival were detected by flow cytometry,Caspase 3 activity detection kit and colony formation assay,respectively.Cell survival curve was stimulated by the multi-target single-hit model.Based on the starBase v2.0 online analysis,the luciferase reporter gene assay,RNA-binding protein immunoprecipitation assay (RIP) and Real-time PCR assay were applied to verify the relationship between CCAT1 and miR-130b-3p.Results CCAT1 expression was up-regulated (t=6.322-8.555,P＜0.05),but miR-130b-3p expression was down-regulated (t =3.950-18.795,P＜ 0.05) in the radiation-resistant pancreatic cancer tissues,pancreatic cancer cell lines and 2 Gy-irradiated PANC-1 cells.When the CCAT1 silenced PANC-1 cells were irradiated with 2 Gy,cell survival fraction decreased (t=2.929,5.047,5.234,5.125,P＜0.05),apoptosis rate and Caspase 3 activity increased (t=6.953,6.836,P＜0.05).CCAT1 could selectively regulate miR-130b-3p expression.Inhibition of miR-130b-3p expression could enhance PANC-l cell survival (t =4.564,6.736,8.656,P＜0.05),but reduced apoptosis rate (t=5.234,P＜0.05) and Caspase 3 activity (t=10.440,P＜0.05).Conclusions Silencing CCAT1 promotes the expression of miR-130b-3p and enhances radiosensitivity of PANC-1 cells.

Mechanism of Silencing HOXC8 Increases Radiosensitivity of A549 Cells / 中华放射肿瘤学杂志

Objective@#To investigate the effect and potential mechanism of HOXC8 on the radiosensitivity of non-small cell lung cancer cell line A549, aiming to provide novel ideas for clinical combined treatment.@*Methods@#The A549 cells with stable knockdown of HOXC8 were constructed by using lentivirus and validated by qPCR and Western blot. The radiosensitivity of A549 stable cell line was assessed by plate clone formation assay. The expression levels of TGF-β1 and the proteins in the downstream signal pathway after knockdown of HOXC8 were detected by Western blot.@*Results@#The A549 cells with stable knockdown of HOXC8 were successfully constructed. The viability and clonogenic capacity of A549 cells were significantly reduced after silencing HOXC8. Silencing HOXC8 also increased the sensitivity of A549 cells to radiotherapy and significantly inhibited the expression of TGF-β1 and p-Smad2/3 proteins in the downstream signaling pathway.@*Conclusion@#Silencing HOXC8 can increase the sensitivity of A549 cells to radiotherapy probably by inhibiting TGF-β1 signaling transduction. HOXC8 might play an important role in A549 cells.

Predictors of virological response in HBeAg-positive chronic hepatitis B patients treated with adefovir dipivoxil / 中华传染病杂志

Objective To investigate the predictive factors of virological response in HBeAg-positive chronic hepatitis B (CHB)patients treated with adefovir dipivoxil (ADV).Methods A total of 203 HBeAg-positive CHB patients treated with ADV (Mingzheng)10 mg once daily for 48 weeks were recruited.The gene polymorphisms at positions-238 and-308 in tumor necrosis factor (TNF)-α promoter region were determined by the restriction fragment length polymorphism assay of products amplified using polymerase chain reaction (PCR-RFLP).The serum levels of TNF-a at baseline were measured by enzyme linked immunosorbent assay (ELISA).Hepatitis B virus (HBV)genotypes were tested by real-time fluorescent quantitative PCR and HBV subgenotypes were tested by HBV S gene sequencing.Factors related to ADV response were determined by Logistic regression analysis.Results The HBV DNA negative rate,alanine aminotransferase (ALT)normalization rate,HBeAg loss rate and seroconversion rate,and combined response rate at week 24 and 48 of treatment in 203 patients were 31.5% (64/203),59.1% (120/203),15.8% (32/203),8.9% (18/203),13.3% (27/203)and 58.6% (119/203),78.3% (159/203),29.6% (60/203),16.7% (34/203),25.6% (52/203),respectively.HBV DNA negative rate at week 24 was higher in patients with HBV genotype B,that was higher in patients with TNF-α-308G/A genotype,and that was higher in patients with higher baseline ALT level or lower baseline HBV DNA level [OR = 0.405,95 % CI (0.191 - 0.859),P =0.019;OR=0.292,95%CI(0.132-0.643),P=0.002;OR=0.933,95%CI(0.989-0.997),P＜0.01 ;OR=2.089,95%CI (1.412-3.092),P＜0.01].Meanwhile,HBV DNA negative rate at week 48 were higher in patients with higher HBV DNA negative rate at week 24 or higher baseline ALT level [OR=0.029,95%CI(0.007-0.126),P＜0.01;OR= 0.995,95%CI(0.991-0.999),P=0.016].Conclusions HBV genotype,TNF-α-308 genotype,baseline levels of ALT and HBV DNA are predictors of virological response at week 24 in HBeAg-positive CHB patients treated with ADV.And the HBV DNA negative rate at week 24 and baseline ALT level are predictors of virological response at week 48.

Polymorphism of TNFα-308 and mutations of HBV C region in patients with chronic HBV infections / 中华临床感染病杂志

Objective To investigate the association between polymorphism of TNFα-308 and mutations of HBV C region in patients with chronic HBV infection. Methods Ninety-five patients with chronic HBV infection were recruited in the study. The single nucleotide polymorphism(SNP) of TNFα-308 Was determined by restriction fragment length polymorphism(RFLP). Mutations of nt1762/1764, nt1896, nt1899, nt1862, as60, aa87 and as97 in HBV C region were detected by direct sequencing after PCR amplification. Mutations of the above points among different genotypes were compared by Fisher's exget test. Results Three different genotypes G/G(63/95, 66.3%), G/A(28/95, 29.5%) and A/A(4/95, 4.2%) were found in TNFα-308 site. The rates of mutations of aa87 and aa97 points in patients with G/G, G/A and A/A genotype were 39.3%(24/61), 11.5%(3/26) and 50.0% (2/4), respectively(F=7.658, P<0.05);while the mutation rates of nt1762/1764, nt1896, nt1864, nt1899 and aa60 were of no statistical significance among different genotypes(F=0.669, 1.542, 1.123, 2.420 and 0.966, P>0.05). Conclusion Compared with G/G genotype, antigenicity of HBV may be more stable in patients with TNFα-308 G/A genotype, which is beneficial to HBV clearance.

Predictive factors for serological response in HbeAg-positive chronic hepatitis B patients with adefovir dipivoxil treatment / 中华临床感染病杂志

Objective To investigate the predictive value of TNFα,ALT,HBV DNA loads and HBV serological markers in response to adefovir dipivoxil (ADV) treatment for patients with chronic hepatitis B(CHB).Methods Two hundred and three HBeAg.positive CHB patients were administered with ADV 10 mg/d for 48 weeks.HBV serological markers and TNFα at the baseline were determined by enzyme linked immunosorbent assay(EUSA),and HBV DNA loads were detected by PCR.Logistic regression was used to identify predictive factors for serological response at 48th week after the treatment.Results The rates of HBV DNA clearance,ALT normalization,HBeAg lOSS,HBeAg seroconversion and response at 24th week were 31.5%(64/203),59.1%(120/203),15.8%(32/203).8.9%(18/203)and 13.3%(27/203)respectively,while those at 48th week were 58.6%(119/203),78.3%(159/203),29.6%(60/203),16.7%(34/203)and 25.6%(52/203),respectively.Patients who achieved HBeAS loss at 48th week were found to have higher rates of HBV DNA clearance.HBeAg loss and seroconversion at 24th week and higher TNFα at baseline(P=0.017,0.ooI,0.029 and 0.040),while those who achieved HBeAg seroconversion at 48th week were found to have higher rate of HBeAg seroconversion at 24th week.and lower baseline HBV DNA loads(P=0.000 and 0.004).Conclusion For HBeAg.positive CHBpatients with ADV treatment,the rate of HBV DNA clearanee,HBeAg loss and seroeonversion at 24th week and TNFα at baseline may be used to predict the rate of HBeAg 1088 at 48th week:the rate of HBeAgseroconversion at 24th week and baseline HBV DNA loads may be used to predict the rate of HBeAgseroeonversion at 48th week.