1.Effect of piperine on AC/PKA/CREB signaling pathway on hippocampal neuron apoptosis in rats with cerebral infarction
Deyan LI ; Zongwen HUANG ; Yadan WU ; Xiuhong CHEN
Chinese Journal of Immunology 2024;40(12):2537-2542
Objective:To investigate the effect and of piperine(PIP)on nerve injury in rats with cerebral infarction.Meth-ods:The rat model of cerebral infarction was prepared by modified thread embolism method.The rats were divided into Sham group,Model group,piperine group(PIP group,20 mg/kg piperine),piperine+PKA inhibitor group(PIP+H-89 group,20 mg/kg PIP+5 mg/kg H-89).The rats were scored for neurological impairment,the volume of cerebral infarction,neuronal damage and neuronal apoptosis were observed,and the levels of cAMP,IL-1β and IL-6 in hippocampus and the protein expressions of GFAP,NSE,AC6,PKA,p-CREB,CREB and BDNF were detected.Results:Compared with Sham group,Model group had large cerebral infarction,cerebral in-farction volume,the neurological deficit score,neuronal apoptosis rate,the levels of IL-6 and IL-1β in hippocampus,and number of GFAP positive cells,the positive expression rates of NSE protein increased,the number of Nissl bodies,and the expressions of cAMP,AC6,PKA,p-CREB and BDNF proteins in hippocampus decreased(P<0.05);compared with Model group,cerebral infarc-tion volume,the neurological deficit score,neuronal apoptosis rate,the levels of IL-6 and IL-1β in hippocampus,and number of GFAP positive cells,the positive expression rates of NSE protein in the PIP group decreased,the number of Nissl bodies,and the ex-pressions of cAMP,AC6,PKA,p-CREB and BDNF proteins in hippocampus increased(P<0.05);the effect of piperine in improving nerve injury in rats with cerebral infarction was able to be reversed by H-89.Conclusion:PIP can inhibit the apoptosis of hippocampal neurons,and improve the neural injury induced by cerebral infarction by activating AC/PKA/CREB signal pathway.
2.Effects of probiotics on intestinal flora, intestinal function and T lymphocyte level in patients with cervical cancer after radiotherapy
Ting CHAI ; Songjie ZHANG ; Rui SONG ; Yanli DUANMU ; Jinjin YUAN ; Ge HOU ; Alan CHU ; Yangyang HUANG ; Shijia LIU ; Zongwen LIU
Chinese Journal of Primary Medicine and Pharmacy 2023;30(7):1012-1017
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.
3.Effect of LncRNA UCA1 on radiosensitivity of lung cancer cells and its mechanism
Cheng WANG ; Zongwen LIU ; Ge HOU ; Jun YANG ; Yangyang HUANG
Chinese Journal of Radiation Oncology 2020;29(4):289-293
Objective:To evaluate the effect of long-chain non-coding RNA (LncRNA) UCA1 on the proliferation, apoptosis and radiosensitivity of lung cancer cell and to explore the underlying mechanism.Methods:qRT-PCR was used to detect the expression of UCA1 and miR-513a-5p in lung cancer cell A549, H1299 and normal human lung cell HBE. The si-con group (transfected si-con), si-UCA1 group (transfected si-UCA1), miR-513a-5p group (transfected miR-513a-5p mimics), miR-NC group (transfected miR-NC), IR+ si-con group (transfected si-con, and irradiated), IR+ si-UCA1 group (transfected miR-NC and irradiated), IR+ miR-513a-5p group (transfected miR-513a-5p mimics and irradiated), IR+ miR-NC group (transfected miR-NC and irradiated), IR+ si-UCA1+ anti-miR-513a-5p group (co-transfected si-UCA1, anti-miR-513a-5p and irradiated) were transfected into the A549 and H1299 cells by liposome method, and then the cells in certain groups were subject to 4Gy irradiation. The cell proliferation of each group was detected by MTT assay. The sensitivity enhancement ratio was assessed by clone formation assay. The cell apoptosis of each group was detected by flow cytometry. The fluorescence activity of each group was detected by dual-fluorescein gene detection assay.Results:Compared with human normal lung cell HBE, the expression levels of UCA1 were significantly up-regulated in the lung cancer cell A549 and H1299(both P<0.05), whereas those of miR-513a-5p were significantly down-regulated (both P<0.05). Inhibition of UCA1 and overexpression of miR-513a-5p significantly inhibited cell proliferation, promoted cell apoptosis and increased the sensitivity of radiation exposure of A549 and H1299(sensitivity enhancement ratio=1.897, 2.146 and 1.615, 1.872). miR-513a-5p could suppress the fluorescence activity of wild-type UCA1 cells, and UCA1 could negatively regulate the expression of miR-513a-5p. Inhibition of miR-513a-5p could reverse the enhancement effect of inhibiting UCA1 upon the radiosensitivity of lung cancer cells. Conclusions:Inhibition of LncRNA UCA1 can enhance the sensitivity of radiation exposure to lung cancer cells. The mechanism may be related to the targeted inhibition of miR-513a-5p.
4.Radiosensitivity of lncrna linc00909 targeting mir-548-3p on colorectal cancer cells
Ge HOU ; Chenhu XIAO ; Xiaojuan CHEN ; Yangyang HUANG ; Cheng WANG ; Ting CHAI ; Rui SONG ; Jinjin YUAN ; Zongwen LIU
Chinese Journal of Radiation Oncology 2020;29(12):1110-1117
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.
5.Isolation and culture of adipose-derived mesenchymal stem cells from inbreed line miniature pig of Wuzhishan and their biological characteristics.
Denggao HUANG ; Hui CAO ; Shunlan WANG ; Linlin ZHENG ; Zongwen CHEN ; Xiaohong WEN ; Shufang ZHANG ; Yang XIANG ; Yuanhui GAO
Journal of Central South University(Medical Sciences) 2019;44(3):297-306
To establish a method for isolation, culture and identification of adipose-derived mesenchymal stem cells (ASCs) from the inbreed line miniature pig of Wuzhishan (ILMW).
Methods: A total of 100 g adipose tissues were obtained from subcutaneous tissues of neck in six-month old healthy ILMW (3 samples, male). ASCs from ILMW (ILMW-ASCs) were isolated from adipose tissues through 0.1% collagenase digestion. The cells at the 3rd, 5th, 8th, 13th passages were collected. Cell morphology, size, phenotype, cell cycle, and apoptosis were monitored. Cell differentiation was induced and cell proliferation curve was drawn.
Results: The ILMW-ASCs, fibroblast-like or whirlpool-like, began the adherence at 36 h and entered a logarithmic phase in the 5th day. Eighty percent of them were fused in the 7th day. The average diameter and volume of ILMW-ASCs were (17.00±0.54) µm and (2.58±0.24)×10-9 L, respectively. The expressions of CD29, CD44 and CD90 were positive, and there was no significant difference between the different passages (all P>0.05). The expressions of CD45, CD8a and HLA-DR were increased with the increase in passages after the 3th passage (all P<0.05). The adipogenic induction of ILMW-ASCs was observed by positive oil red O staining, and the osteogenic induction of ILMW-ASCs was determined by positive alizarin red staining. Apoptosis and senescence occurred in the 13 passage of ILMW-ASCs, and the proportion of S phase of cell cycle was lower than that in lower passages (all P<0.05).
Conclusion: ILMW-ASCs are one of the best choice for porcine ASCs, which might provide a source of candidate stem cells for therapy of large animal disease models and tissue or organ repairment.
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Swine, Miniature
6.Efficacy of Rosuvastatin Combined with Danhong Injection in the Treatment of Unstable Angina Pectoris and Influence on Serum Levels of TNF-a, hs-CRP and IL-6
Qian LU ; Lewen LI ; Zongwen HUANG
China Pharmacist 2018;21(1):109-112
Objective:To discuss the efficacy of rosuvastatin combined with Danhong injection in the treatment of unstable angina pectoris (UAP) and the influence on serum levels of TNF-α, hs-CRP and IL-6.Methods:Totally 72 patients with UAP were random-ly divided into the control group (n=36) and the observation group (n=36).All the patients received the conventional therapy .The control group was given rosuvastatin tablets ,10 mg,po, qd on the basis of the conventional therapy , while the research group was trea-ted with Danhong injection 30ml,ivd, qd on the basic of the control group .All the patients were treated for 2 weeks.The improvements of clinical symptoms and electrocardiogram , and the adverse reactions before and after the treatments were observed .The frequency of onset and duration before and after the treatment were recorded .The levels of blood lipid , TNF-α, hs-CRP and IL-6 before and after the treatment were detected .Results: The total effective rate of the observation group was 91.67%, which was significantly higher than that (72.22%) of the control group (P<0.05).The seizure frequency and duration of the research group were significantly low -er than those of the control group (P<0.05).After the treatment, the lipid indices such as TG, TC and LDL-C, and serum levels of TNF-α, hs-CRP and IL-6 of the two groups decreased significantly , while HDL-C significantly increased , and the differences were sta-tistically significant (P<0.05).The levels of TG, TC, LDL-C, TNF-a, hs-CRP and IL-6 were significantly lower in the observation group than those in the control group , while HDL-C was significantly higher than that in the control group (P<0.05).Conclusion:Danhong injection and rosuvastatin can effectively improve the clinical symptoms of UAP patients , reduce angina frequency and dura-tion, and decrease serum levels of TNF-a, hs-CRP and IL-6 to inhibit inflammation , which is safe and effective , and worthy of clinical application.
7.Correlation analysis of two different techniques for the score of sleep breathing events: cardiopulmonary coupling vs polysomnography
Jing FENG ; Huijuan WU ; Zongwen WANG ; Kun CHEN ; Bei HUANG ; Zhongxin ZHAO
Chinese Journal of Neurology 2017;50(8):606-612
Objective To explore the relationship between the respiratory disturbance index (RDI) detected by cardiopulmonary coupling (CPC) and all parameters of polysomnography (PSG), and to analyze the correlation of the two different analytical techniques for the interpretation of sleep breathing events.Methods In this case-control study, the patients′ CPC and PSG were simultaneously recorded at the Sleep Clinic at Changzheng Hospital from Janunary 2016 to December 2016.Effective recordings were obtained from 292 patients (male/female: 173/119).According to PSG, these patients were divided into two groups: obstructive sleep apnea hyponea syndrome (OSAHS) group (n=173, aged (50.1±13.3) years) and non-OSAHS group (n=119, aged (50.5±11.7) years).All the PSG parameters and the RDI detected by CPC were compared between the two groups.Pearson correlation was used to analyze the relationship between the CPC-RDI and polysomnography parameters, and Bland-Altman chart was used to test the correlation of the two methods.The receiver operator characteristic curve was used to research the value of CPC-RDI in diagnosis of OSAHS.Results The OSAHS group showed higher levels of total sleep time (TST, (445.94±82.64) min), apnea times ((108.16±35.70)/h), hypopnea rate ((55.62±17.44)/h), apnea hyponea index (AHI, (22.78±20.73)/h), Epworth Sleeping Scale (ESS) scores ((11.21±5.30) scores), CPC-RDI ((32.98±22.19)/h) compared with the non-OSAHS group ((417.21±96.10) min,t=2.730;(7.89±5.41)/h,t=30.384;(11.05±2.23)/h, t=27.709;(5.51±3.11)/h,t=9.014;(7.61±2.29)scores,t=6.973;(11.16±8.63)/h,t=10.205, all P<0.01), and a significant decrease in N1 latency ((14.79±9.29) min vs (18.18±8.98)min, t=-3.106), N3% ((6.53±4.95)% vs(8.83±7.29)%,t=-3.212) and the lowest oxygen saturation (SaO2;(77.91±12.21)% vs(92.72±5.17)%, t=-12.479, all P<0.05 respectively).Pearson correlation analysis indicated that RDI was positively correlated with TST, N2 sleeping time, N3 sleeping time, AHI, microarousals index, leg movements index, and ESS scores (P<0.01 respectively).Remarkably, the correlation between CPC-RDI and PSG-AHI was excellent (r=0.801, P<0.01), and CPC-RDI was negatively correlated with lowest SaO2 (r=-0.765, P<0.01).Bland-Altman showed that the points in the limits of agreement accounted for more than 95% of all points.If CPC-RDI is higher than 18.95, the subjects were more likely with OSAHS.The sensitivity was 62.9% and the specificity was 88.7%.In addition, the enhanced low-frequency coupling parameters in the CPC analysis technique can clearly analyze the central respiratory rhythm disturbance.Conclusions CPC technology is an effective assessment technology to diagnose sleep-disordered breathing, and correlates well with AHI detected by traditional PSG.It shows the advantage of screening for central sleep apnea.
8.The value of 64-row helical CT multi-phase enhancement scan combined with angiography in the diagnosis of pulmonary mass
Chao YANG ; Xianlong HUANG ; Hua YANG ; Biqiang LI ; Zhuoyue TANG ; Yuanhao HU ; Zongwen LI ; Xiuyan CHEN
Chongqing Medicine 2015;(7):916-918
Objective To explore the value of 64-row helical CT multi-phase enhancement scan combined with angiography (CTA)in the diagnosis of pulmonary mass.Methods Two hundred and sixty-five patients with pulmonary mass confirmed by pa-thology were checked,analyzed the CT sign of multi-phase enhancement scan and the blood supply of pulmonary mass displayed by CTA.Results Lung cancer was mainly supplied by bronchial arteries,some by body arteries,the feeding arteries display rate of lung cancer group was significantly higher than that of benign disease group(P <0.05).CT enhancement peak value of lung cancer group was significantly higher than that of tuberculoma group,inflammatory pseudotumor group and hamartoma group(P <0.05), but no significant difference between hemanginoma group(P >0.05).Enhancement dynamic curves of lung cancer group was differ-ent from benign lesion groups:Lung cancer without obvious enhancement in pulmonary artery phase,CT value increased rapidly in aorta phase,120 s reached peak,and declined slowly in delay phase;CT value of tuberculoma was increased slowly without obvious peak;CT value of inflammatory increased gradually in pulmonary artery phase,90 s reached the peak;hamartoma was no obvious enhancement;Hemangioma enhanced rapidly after strengthening in the pulmonary artery phase,reached the peak at about 15 s,and then decreased slowly.Conclusion 64-row helical CT multi-phase enhancement scan combined with angiography have important clinical value,which can differentiate malignant mass from benign ones.
9.Involved-field radiotherapy versus elective nodal irradiation in combination with concurrent chemotherapy for locally advanced non-small cell lung cancer: A prospective randomized clinical study
Honglian MA ; Yong BAO ; Xiao HU ; Jin WANG ; Weihua WANG ; Kaixin LI ; Yuan LIU ; Han HE ; Zongwen SUN ; Yan WANG ; Tingting ZHUANG ; Jing CHEN ; Fang PENG ; Qichao ZHOU ; Shaomin HUANG ; Zhichun HE ; Li ZHANG ; Xiaowu DENG ; Ming CHEN
Chinese Journal of Radiation Oncology 2012;21(4):315-320
ObjectiveTo evaluate the local failure and the impact on survival by prospectively comparing involved field radiotherapy (IFRT) and elective nodal irradiation (ENI) in combination with concurrent chemotherapy for locally advanced non-small cell lung cancer ( LA-NSCLC ).Methods LANSCLC patients were treated with 2 cycles of carboplatin ( AUC =5 - 6,d1 ) combined with paclitaxel ( 175mg/m2 ),followed assessment without distant metastasis,then randomized into IFRT (45 patients) or ENI (54 patients) arm.IFRT included primary tumor,ipsilateral hilar and positive mediastinal lymph nodes;ENI included the primary lesion,ipsilateral hilar,hilateral mediastinal lymph node drainage and bilateral supraclavicular area.The prescription dose was given as high as possible with V20 ≤35% and spinal cord dose ≤50 Gy,combined weekly paclitaxel 40 mg/m2 concurrent chemotherapy.The Kaplan-Meier method was used to estimate survival data and the log-rank method was used to test distribution of survival time between arms.ResultsThe follow-up rate was 99%.49,29 and 17 patients were followed-up for 1-,2-and 3-year,respectively.More patients from group IFRT received >60 Gy than ENI (49% vs.26%,x2 =5.59,P =0.018 ).The local failure rates were 29% and 36%,respectively ( x2 =0.46,P =0.497 ).The 1-,2-and 3-year local tumor progression-free survival rates were 76%,69%,65% and 80%,53%,49% ( x2 =0.74,P =0.389),respectively; the 1-,3-and 5-year overall survival rates were 80%,41%,33% and 69%,32%,13% (x2 =3.97,P =0.046),respectively.There were no significant differences in acute and late toxicities between the arms ( x2 =3.910 - 0.155,P =0.142 - 0.925 ).ConclusionsIFRT improved radiation dose and survival rate and did not increase the failure of elective lymph node region compared with ENI.The toxicities were no differences between IFRT and ENL Further investigation with big size sample is warranted.
10.CT morphometric assessment of the acromion and clavicle hook plate
Tao CHEN ; Meng YE ; Yuanqing GUO ; Chuangxin HUANG ; Guowei LI ; Jiaying LAI ; Zongwen HUANG
Chinese Journal of Orthopaedic Trauma 2012;14(1):11-14
Objective To measure the morphological parameters of the acromion with CT and to analyze their match with the hook plate. Methods From October 2009 to February 2010,spiral CT scanning (with Somatom Emotion16) and three-dimensional reconstruction of bilateral shoulders were conducted in 61 Chinese subjects.They were 24 men and 37 women,aged from 20 to 83 years (average,45.2 years).The thickness,length and width of the acromion were measured and the subacromial shape was observed with software of the CT system to analyze the match between the hook plate and the acromion.Results The mean thickness of the acromion was 0.85 ± 0.13 cm in all subjects,0.94 ± 0.12 cm in males and 0.79 ± 0.10 cm in fe males,with a significant difference between males and females ( t =2.382,P =0.202).The mean acromion length was 2.08 ± 0.20 cm in all subjects,2.09 ± 0.21 cm in males and 2.06 ± 0.18 cm in females,with no significant difference between males and females( t =1.541,P =0.129).The mean acromion width was 3.81 ± 0.52 cm in all subjects,4.34 ± 0.32 cm in males and 3.47 ± 0.25 cm in females,with a significant difference between males and females ( t =2.296, P =0.025 ). Conclusions The acromial morphology varies significantly between genders in Chinese population,particularly in the thickness and width.It is,therefore,necessary to modify the morphological parameters of the hook plate to match better the gender difference in Chinese population.It is optimal that the hook plate should allow individualized pre-moulding to enhance its therapeutic efficacy.

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