3.A novel hydrogen sulfide donor modulates Kir6.2 via the cyclic guanosine monophosphate- dependent protein kinase signaling pathway in one side hippocampus of epileptic rats
Danqiong LIU ; Ziting ZHU ; Sheng DU ; Peiye CHEN ; Manru GUO ; Yilong ZHANG ; Yanmei SU ; Guanqi WANG ; Pingyi XU ; Xiaoqin ZHU
Chinese Journal of Neurology 2021;54(3):242-250
Objective:To explore the effect of hydrogen sulfide (H 2S) on modulating the subunit Kir6.2 of adenosine triphosphate sensitive potassium channels via the cyclic guanosine monophosphate-dependent protein kinase (cGMP/PKG) signaling pathway in epileptic rat models. Methods:Sixty adult male SD rats were randomly divided into the following six groups (10 rats in each group) by random number table method: control, epileptic, H 2S donor, H 2S donor+epileptic, KT5823 (one inhibitor of the cyclic guanosine monophosphate-dependent protein kinase)+H 2S donor+epileptic, and glibenclamide (one inhibitor of the adenosine triphosphate sensitive potassium channels)+H 2S donor+epileptic groups. Except the control group, SD rats were intraperitoneally injected with plentylenetetrazole to make the kindling models and their behaviours were recorded including the latency period, the grade, and the duration of the first epileptic seizure according to the Racine′s standard. The waveforms of electroencephalogram (EEG) in hippocampus were also recorded during the seizure. The mRNA and protein levels of PKG and Kir6.2 in hippocampus were evaluated by Western blotting and quantitative real-time polymerase chain reaction, and the hippocampal concentrations of cGMP and phosphorylation of cyclic guanosine monophosphate-dependent protein kinase (p-PKG) were detected by enzyme linked immunosorbent assay. Results:Rats in the epileptic group showed Ⅳ-Ⅴ grade of epileptic seizure [4.500 (4.000, 4.875)], short latency period [(10.37±8.21) min] but long duration [(69.50±24.37) s] of seizure. Compared to the epileptic group, rats in the H 2S donor group showed Ⅱ-Ⅲ grade of epileptic seizure ( P=0.004), significantly longer latency period ( P<0.001), and shorter duration of seizure ( P<0.001). Compared to the H 2S donor+epileptic group, rats in the KT5823+H 2S donor+epileptic group showed Ⅲ-Ⅳ grade of epileptic seizures, significantly shorter latency period ( P<0.001), and longer duration of seizure ( P<0.001). The results of EEG showed that the wave patterns in the epileptic group were spike or sharp waves and the amplitudes were largest [(190.570±23.590) μV]. Compared with the epileptic group, amplitudes were reduced ( P<0.001) in the H 2S donor+epileptic group. PKG mRNA and PKG protein were expressed differently among all groups (PKG mRNA: n=5, H=26.714, P<0.001; PKG protein: n=5, F=30.597, P<0.001). Compared with the control group, the expression of both PKG mRNA and PKG protein was decreased (PKG mRNA: 1.000±0.001 vs 0.782±0.064, P=0.023; PKG protein: 0.550±0.037 vs 0.145±0.020, P=0.042) in the epileptic group. Besides, Kir6.2 mRNA and Kir6.2 protein were expressed differently among all groups (Kir6.2 mRNA: n=5, H=27.761, P<0.001; Kir6.2 protein: n=5, F=60.659, P<0.001). Compared with the control group, the expression of both Kir6.2 mRNA and Kir6.2 protein was decreased (Kir6.2 mRNA: 1.000±0.001 vs 0.897±0.033, P=0.004; Kir6.2 protein: 0.384±0.035 vs 0.215±0.016, P=0.024) in the epileptic group. And the concentrations of cGMP and p-PKG were decreased (cGMP: P<0.001; p-PKG: P<0.001) in the epileptic group. The results in the H 2S donor+epileptic group were up-regulated (PKG mRNA: P=0.047; PKG protein: P<0.001; Kir6.2 mRNA: P=0.011; Kir6.2 protein: P<0.001; cGMP: P<0.001; p-PKG: P<0.001) compared with the epileptic group. However, the results in the KT5823+H 2S donor+epileptic group were down-regulated (PKG mRNA: P=0.015; PKG protein: P=0.027; Kir6.2 mRNA: P=0.013; Kir6.2 protein: P=0.017; cGMP: P=0.005; p-PKG: P<0.001) compared with the H 2S donor+epileptic group. Conclusion:A possible mechanism is that H 2S prevents the epileptic seizure from modulating the subunit Kir6.2 of ATP sensitive potassium channels via the cGMP/PKG signaling pathway.
4.Reproducibility of intravoxel incoherent motion in body DWI technique for normal lumbar disc scanning
Yuan WANG ; Sheng ZHOU ; Wenqi WANG ; Xiaofei CHEN ; Fuwen DONG ; Yanmei QI
Journal of Practical Radiology 2019;35(6):970-973
Objective To investigate the reproducibility of magnetic resonance intravoxel incoherent motion in body diffusionG weighted imaging (IVIMGDWI)for normal lumbar disc scanning.Methods 50 healthy volunteers were enrolled with informed consent,30 males and 20 females,2 5.20±2.04 years old.Using 3.0T MR on the lumbar spine,the sagittal T1 WI,the sagittal,axial T2 WI and sagittal IVIMGDWI sequences were scanned once,then the second sagittal IVIMGDWI sequence was scanned after 4 hours.The IVIMGDWI sequence used 10 b values (0,10,20,40,60,80,100,200,400,600 s/mm2 ).The discs were graded according to the Pfirrmann grading standard.The ADCfast ,ADCslow ,and f values of each intervertebral disc were measured by two doctors at the postGprocessing workstation.Paired tGtest was used to analyze whether there was a difference between the two scans.The intraGgroup correlation coefficient (ICC)was used to analyze the consistency of the ADCfast ,ADCslow ,and f values of the two acquisitions (P<0.05)and the consistency of the ADCfast ,ADCslow,and f values between the different doctors (P<0.05).The IVIMGDWI imaging was evaluated to measure the repeatability of normal lumbar discs. Results Of the 50 healthy volunteers,230 intervertebral discs matched the criteria(Pfirrmann gradeⅠandⅡ).The ADCslowvalue between the two scans was significantly different (t=2.460,P<0.05),and the differences in ADCfast and f values were not significant (t=-0.418,1.273,P>0.05). The consistency of ADCfast ,ADCslow ,and f values for the two scans were generally (ICC=0.478,0.306,0.316,P<0.05 ).Different observers had good consistency in the measurement of interverG tebral disc ADCfast ,ADCslow,and f values (ICC=0.929,0.909, 0.9 1 1 ,P<0.05).Conclusion The IVIMGDWI imaging has good consistency in the measurement of normal lumbar disc between different observers.The consistency of IVIMGDWI in two scans of normal lumbar intervertebral discs is general,which may be due to the time interval between the two scans.Because ADCslow represents the diffusion of water molecules in tissues,the microenvironment in the lumbar intervertebral disc has changed,resulting in the difference of ADCslow value.As for the problem that different scanning time may lead to the change of IVIMGDWI data,we will study it further.
5.LncRNA TCF7 promotes malignant biological behavior of lung cancer cells by regulating miR-29 and activating JAK/STAT2 signaling pathway
Gaowa JIN ; Huaqing WANG ; Sheng HU ; Yanmei GAO
International Journal of Biomedical Engineering 2018;41(6):527-533
Objective To investigate the malignant biological behavior and mechanism of LncRNA TCF7 in lung cancer cells by regulating miR-29 and activating JAK/STAT2 signaling pathway.Methods qPCR was used to detect the expression of TCF7 and miR-29 in lung cancer tissues and different lung cancer cell lines.The relationship between TCF7 and clinicopathological data of lung cancer patients was analyzed.The dual luciferase reporter assay was used to detect the interaction between TCF7 and miR-29.MTT proliferation assay and Transwell invasion assay was used to detect the proliferation and invasion of lung cancer cells after inhibition of TCF7,respectively,and to analyze the relevant recovery after the overexpression of miR-29.The expression of JAK/STAT2 signaling pathway protein was detected by Western Blotting after TCF7 inhibition.The effect of TCF7 on tumor formation in vivo was detected by in vitro tumor formation assay in nude mice.Results Compared with other lung cancer cell lines,A549 cells had the highest expression of TCF7 and miR-29.The expression of TCF7 was associated with the pathological stage of lung cancer and lymph node metastasis,in which TCF7 was positively correlated with cancer stage and lymph node metastasis.The dual luciferase assay confirmed that TCF7 can specifically bind to the target of miR-29,and regulate the expression and activity of miR-29.The inhibition of the expression of TCF7 can promote the proliferation and invasion of lung cancer cells.After inhibiting the expression level of miR-29,the proliferation and invasion ability of lung cancer cells were partly restored.After inhibiting the expression of TCF7,JAK/STAT2 signaling pathway was activated accordingly.Compared with the non-small carcinoma group,the average tumor volume and mass of the transplanted tumor in the TCF7-siRNA group were reduced.Conclusions TCF7 can regulate the expression of miR-29 and affect the proliferation and invasion of lung cancer cells through JAK/STAT2 signaling pathway.
6.Trajectories and influencing factors of post-traumatic stress disorder in women with multiple in vitro fertilization embryo transfer failures
Yanhua SUN ; Jing ZHAO ; Yanmei HU ; Xia WANG ; Xiaohui SHENG ; Xiaoyue ZHANG
Chinese Journal of Practical Nursing 2024;40(2):129-136
Objective:To explore the potential categories of post-traumatic stress disorder (PTSD) trajectories in women with multiple in vitro fertilization-embryo transfer (IVF-ET) failures, and to analyze the effects of different demographic characteristics and psychological factors on the potential categories of PTSD trajectories.Methods:This was a prospective empirical research, from May 2021 to October 2022, women with IVF-ET failure ≥ 2 times in the reproductive department of Shanghai First People′s Hospital from May 2021 to October 2022 were selected as the research objects. Post-traumatic stress disorder civilian version scale was used for 4 follow-ups at 3 d (T1), 10 d (T2), 20 d (T3) after the last transplantation failure and 3 d before the next transplantation cycle (T4). Telephone follow-up and online follow-up were combined to obtain the PTSD level at 4 time points. Potential categories of PTSD score trajectories at four time points were identified using a latent category growth model, and analyze influencing factors using unordered multi classification logistic analysis.Results:Totally 196 IVF-ET women were admitted, aged (29.42 ± 4.13) years. Three PTSD trajectories were fitted in this study, including 82 cases (42%) in non-PTSD group, 61 cases (31%) in mild PTSD group and 53 cases (27%) in elevated PTSD group. Logistic regression analysis showed that age, education level, fertility pressure and marital adjustment level were the predictors of PTSD trajectory in women with multiple IVF-ET failures. Compared with the non-PTSD group, women aged ≥35 years, with lower education level and marital adjustment level were more likely to enter the elevated PTSD group ( OR=4.570, 8.540, 0.949, all P<0.05). Women aged 35 years and with greater reproductive pressure were more likely to enter the mild PTSD group ( OR=3.871, 1.063, both P<0.05). Conclusions:There is group heterogeneity in the trajectories of PTSD in women with multiple IVF-ET failures in the next transplantation cycle. Old age, low education level, high fertility pressure and poor marital adjustment can predict the trajectories of PTSD. Fertility stress and marriage adjustment are changeable variables. Medical staff can relieve women′s fertility pressure through health education and mindfulness intervention, promote a good state of marriage adjustment, and minimize the adverse effects of PTSD on the next cycle of conception.
7.Role of mammalian target of rapamycin in synaptic plasticity of entorhinal area-hippocampal formation in rats with inflammatory pain
Dan LYU ; Zhun WANG ; Wenchen JIANG ; Mingwei SHENG ; Yanmei YANG ; Jing LUAN ; Ying ZHENG ; Yongjin HE
Chinese Journal of Anesthesiology 2018;38(11):1343-1346
Objective To evaluate the role of mammalian target of rapamycin (mTOR) in the synaptic plasticity of entorhinal area-hippocampal formation in rats with inflammatory pain.Methods Twenty-four healthy adult male Sprague-Dawley rats,weighing 180-240 g,were divided into 4 groups (n =6 each) by using a random number table method:control group (group C),inflammatory pain group (group IP),dimethyl sulfoxide (DMSO) group and mTOR inhibitor rapamycin group (group R).Inflammatory pain model was established by subcutaneous injection of 50 μl bee venom into the plantar surface of the left hindpaw.The equal volume of normal saline was subcutaneously injected into the plantar surface of the left hindpaw in group C.In group DMSO,2% DMSO was administered by intragastric gavage for 3 days,1 ml per day,and the inflammatory pain model was established at 1 h after administration on 3rd day.In group R,rapamycin was administered by intragastric gavage for 3 days,1 ml per day,and the inflammatory pain model was established at 1 h after administration on 3rd day.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 2 h after establishing the model.The rats were sacrificed after measurement of the pain threshold,and hippocampi were removed to prepare hippocampal slices.Hippocampal CA1 region and dentate gyrus (DG region) were located with an inverted microscope.Planar microelectrode array technique was used to record the number of channels and the standardized amplitude of evoked effective field excitatory postsynaptic potentials (fEPSPs) (fEPSPs amplitude>20% of the baseline value) at different stimulus intensities.Results Compared with group C,MWT was significantly decreased,TWL was shortened,the number of effective fEPSP channels at different stimulus intensities was increased,and the amplitude of standardized fEPSPs in hippocampal DG and CA1 regions was increased in group IP (P<0.05 or 0.01),and no significant change was found in the parameters mentioned above in group R (P>0.05).Compared with group IP,MWT was significantly increased,TWL was prolonged,the number of effective fEPSP channels at different stimulus intensities was decreased,and the amplitude of standardized fEPSPs in hippocampal DG and CA1 regions was decreased in group R (P<0.05 or 0.01),and no significant change was found in the parameters mentioned above in group DMSO (P>0.05).Conclusion mTOR is involved in the changes in the synaptic plasticity of entorhinal areahippocampal formation in rats with inflammatory pain.
8.Application of gene screening technology in screening common newborn genetic diseases
Hu HAO ; Wei ZHOU ; Congcong SHI ; Sitao LI ; Yanmei MA ; Xia GU ; Hui XIONG ; Bingqing LIU ; Yao CAI ; Guo-Sheng LIU ; Zhichun FENG ; Xin XIAO
Chinese Journal of Applied Clinical Pediatrics 2020;35(22):1712-1717
Objective:To detect the genes of common genetic diseases in newborns with the high-throughput sequencing technology based on target gene capture, to study the incidence rate of such diseases, the carrying rate and variant types of pathogenic mutations related to such diseases, and to explore the application value of the high-throughput sequencing technology in screening genetic diseases of newborns.Methods:The heel blood of 1 793 newborns born in Guangdong province from June 2019 to April 2020 were collected, and the exon regions of 138 common genetic disease-related genes in neonates were detected using the high-throughput sequencing technology based on target gene capture.The pathogenicity of the mutations was interpreted according to the " Classification Criteria and Guidelines for Genetic Variation(2017)" , in which known disease and probable disease were considered as positive mutations.The positive mutations were verified by Sanger sequencing technology, and the test results were analyzed with statistical methods.Results:Among the 1 793 newborns, 978 were male and 815 were female.A total of 158 positive cases were screened(8.81%), and 11 positive diseases were detected.Among the positive diseases, there were 41 cases(2.29%)of autosomal recessive deafness type 1A, 40 cases(2.23%)of Gilbert syndrome or Crigler-Najjar syndrome, and 33 cases(1.84%)of glucose-6-phosphate dehydrogenase deficiency(1.84%), 19 cases(1.06%)of familial hypercho-lesterolemia, 18 cases(1.00%) of sodium taurocholate cotransporter peptide deficiency disease, 2 cases(0.11%)of mitochondrial non-syndromic deafness, 2 cases(0.11%)of Citrin deficiency, 1 case(0.06%)of holocarboxylase synthase deficiency, 1 case(0.06%)of β-thalassemia and 1 case(0.06%)of metachromatic leukodystrophies.Of all studied cases, 972 carried one or more positive mutations, involving 85 kinds of diseases in total.The diseases with a high carrying rate were Gilbert syndrome or Crigler-Najjar syndrome(359 cases, 20.02%), autosomal recessive deafness type 1A(302 cases, 16.84%), and sodium taurocholate cotransport peptide deficiency disease(291 cases, 16.22%). The high-frequency mutation sites were UGT1A1 gene c. 211G> A, GJB2 gene c .109G> A and SLC10A1 gene c. 800C> T. Conclusions:The common genetic diseases detected in neonates from Guangdong province are autosomal recessive deafness type 1A, Gilbert syndrome or Crigler-Najjar syndrome, glucose-6-phosphate dehydrogenase deficiency, familial hypercholesterolemia, and sodium taurocholate cotransport peptide deficiency.There are high-frequency carrying mutation sites in the population.Preliminary genetic screening of common neonatal genetic diseases can accumulate data and experience for the development of newborn genetic screening.
9.Optimization of the Best Proportion of Astragalus membranaceus Injection Combined with Erigeron breviscapus Injection against Cerebral Ischemia-reperfusion Injury in Rats by Baseline Geometric Proportion Increasing and Decreasing Design
Feng TIAN ; Jicao LI ; Shaowei TANG ; Yushi SUN ; Shijia LIN ; Fangyan ZHANG ; Xingliang XIE ; Yanmei SHENG
China Pharmacy 2019;30(14):1885-1889
OBJECTIVE: To screen the best proportion of Astragalus membranaceus injection combined with Erigeron breviscapus injection against cerebral ischemia-reperfusion injury in rats. METHODS: Male SD rats were randomly divided into sham operation group, model group and administration group [different A. membranaceus injection-E. breviscapus injection proportion groups, being A(0 ∶ 10), B(2 ∶ 8), C(4 ∶ 6), D(6 ∶ 4), E(8 ∶ 2), F(10 ∶ 0)groups, set by baseline geometric proportion increasing and decreasing design], with 8 rats in each group. Except for sham operation group, reperfusion injury model of middle cerebral artery occlusion were induced by modified suture method in rats. The each administration group was given relevant medicine intraperitoneally once immediately after inducing model, and then given again after 24 hours (medication interval between the two injections of 30 min). Constant volume of normal saline was given to rats in sham operation group and model group. Forty-eight hours after reperfusion, Longa scoring method was used to evaluate neurological impairment of rats, and neurological impairment score was recorded. Serum content of MDA and activity of SOD were measured by colorimetry assay. TTC assay was used to detect cerebral infraction, and cerebral infarction rate was calculated. Kim’s formula was used to calculate the synergistic index (q) of rats in administration groups. RESULTS: Compared with sham operation group, neurological impairment score and serum content of MDA were increased significantly in model group, while activity of SOD was decreased significantly (P<0.01). The area of cerebral infarction increased significantly, and the rate of cerebral infarction increased significantly (P<0.01). Compared with model group, neurological impairment scores and serum contents of MDA were decreased significantly in group A, B, C, D and E; neurological impairment score of group C was significantly lower than those of group A and F; serum contents of MDA in group B, C, D and E were significantly lower than that of group F (P<0.05 or P<0.01). Activities of SOD in group A, B, C, D and E were increased significantly, and group C was significantly higher than group F (P<0.05 or P<0.01). The cerebral infarction area of rats in each administration group was reduced to varying degrees. The cerebral infarction rates of rats in group B, C, D and E were significantly reduced, and group C and D were significantly lower than group F, while group C was significantly lower than group A (P<0.05 or P<0.01). The q values of group B, C, D and E were 0.90, 1.30, 1.00, 0.70 (neurological impairment score) and 0.79, 1.27, 0.98, 0.82 (cerebral infarction rate). CONCLUSIONS: Different ratios of A. membranaceus injection and E. breviscapus injection have certain protective effects on cerebral ischemia-reperfusion injury model rats, can relieve their neurological deficits, alleviate their oxidative stress and reduce their cerebral infarction areas. The effect of the combination of the two drugs is better than that of single use, and the optimum ratio is 4 ∶ 6.
10.Clinical guideline on first aid for blast injury of the chest (2022 edition)
Zhiming SONG ; Jianming CHEN ; Jing ZHONG ; Yunfeng YI ; Lianyang ZHANG ; Jianxin JIANG ; Mao ZHANG ; Yang LI ; Guodong LIU ; Dingyuan DU ; Jiaxin MIN ; Xu WU ; Shuogui XU ; Anqiang ZHANG ; Yaoli WANG ; Hao TANG ; Qingshan GUO ; Yigang YU ; Xiangjun BAI ; Gang HUANG ; Zhiguang YANG ; Yunping ZHAO ; Sheng LIU ; Lijie TAN ; Lei TONG ; Xiaoli YUAN ; Yanmei ZHAO ; Haojun FAN
Chinese Journal of Trauma 2022;38(1):11-22
Blast injury of the chest injury is the most common wound in modern war trauma and terrorist attacks, and is also the most fatal type of whole body explosion injury. Most patients with severe blast injury of the chest die in the early stage before hospitalization or during transportation, so first aid is critically important. At present, there exist widespread problems such as non-standard treatment and large difference in curative effect, while there lacks clinical treatment standards for blast injury of the chest. According to the principles of scientificity, practicality and advancement, the Trauma Society of Chinese Medical Association has formulated the guidance of classification, pre-hospital first aid, in-hospital treatment and major injury management strategies for blast injury of the chest, aiming to provide reference for clinical diagnosis and treatment.