Directed differentiation of human induced pluripotent stem cells into midbrain / 南方医科大学学报

OBJECTIVE@#To establish an efficient protocol for directed differentiation of human induced pluripotent stem cells (hiPSCs) into functional midbrain dopaminergic progenitor cells (DAPs) in vitro.@*METHODS@#hiPSCs were induced to differentiate into DAPs in two developmental stages. In the first stage (the first 13 days), hiPSCs were induced into intermediate cells morphologically similar to primitive neuroepithelial cells (NECs) in neural induction medium containing a combination of small molecule compounds. In the second stage, the intermediate cells were further induced in neural differentiation medium until day 28 to obtain DAPs. After CM-DiI staining, the induced DAPs were stereotactically transplanted into the right medial forebrain bundle (MFB) of rat models of Parkinson's disease (PD). Eight weeks after transplantation, the motor behaviors of PD rats was evaluated. Immunofluorescence assay of brain sections of the rats was performed at 2 weeks after transplantation to observe the survival, migration and differentiation of the transplanted cells in the host brain microenvironment.@*RESULTS@#hiPSCs passaged stably on Matrigel showed a normal diploid karyotype, expressed the pluripotency markers OCT4, SOX2, and Nanog, and were positive for alkaline phosphatase. The primitive neuroepithelial cells obtained on day 13 formed dense cell colonies in the form of neural rosettes and expressed the neuroepithelial markers (SOX2, Nestin, and PAX6, 91.3%-92.8%). The DAPs on day 28 highly expressed the specific markers (TH, FOXA2, LMX1A and NURR1, 93.3-96.7%). In rat models of PD, the hiPSCs-DAPs survived and differentiated into TH+, FOXA2+ and Tuj1+ neurons at 2 weeks after transplantation. Eight weeks after transplantation, the motor function of PD rats was significantly improved as shown by water maze test (P < 0.0001) and apomorphine-induced rotation test (P < 0.0001) compared with rats receiving vehicle injection.@*CONCLUSION@#HiPSCs can be effectively induced to differentiate into DAPs capable of differentiating into functional neurons both in vivo and in vitro. In rat models of PD, the transplanted hiPSCs-DAPs can survive for more than 8 weeks in the MFB and differentiate into multiple functional neurocytes to ameliorate neurological deficits of the rats, suggesting the potential value of hiPSCs-DAPs transplantation for treatment of neurological diseases.

Effects of manipulator operation task and troubleshooting task on blood pressure indexes of oceanauts / 中华劳动卫生职业病杂志

Objective: To explore the characteristics and rules of blood pressure changes in oceanauts during simulated operation of manipulator and troubleshooting tasks with different difficulty. Methods: In July 2020, 8 deep-sea manned submersible oceanauts, 6 males and 2 females, were selected as objects. In the 1∶1 model of Jiaolong deep-sea manned submersible, the oceanauts performed manipulator operation tasks and troubleshooting tasks with different difficulties, measured the continuous blood pressure of the oceanauts, filled in the NASA Task Load Index (NASA-TLX scale) after the completion of a single mission, and the changes of systolic pressure (SBP), diastolic pressure (DBP), mean arterial pressure (MAP) and mental workload were analyzed. Results: In a single task, the SBP, DBP and MAP of the oceanauts increased first and then decreased. The blood pressure values at the third minute were significantly lower than those at the first minute (P<0.01), and those at the fifth minute were significantly higher than those at the third minute (P<0.01). When performing the same task, compared with the quiet state, SBP, DBP and MAP increased when the oceanauts performed low difficulty, high difficulty, high difficulty+2-back manipulator operation task and troubleshooting task (P<0.05). When the task difficulty was the same, the SBP and MAP of oceanauts performing manipulator operation tasks were higher than those of oceanauts performing troubleshooting tasks (P<0.05). Compared with low difficulty tasks, the scores of NASA-TLX scale for oceanauts performing high difficulty manipulator operationtasks were significantly higher (P<0.05). Compared with the low difficulty task and high difficulty task, the scale score of the high difficulty+2-back troubleshooting task was significantly higher (P<0.05). When the task difficulty was the same, the scale scores of low difficulty and high difficulty manipulator operation tasks were significantly higher than those of troubleshooting tasks (P<0.05). SBP, DBP, MAP of No. 1, No. 3, No. 4, No. 5, and No. 7 oceanauts (all of whom had 6 years of diving) were positively correlated with NASA-TLX scale scores (r>0.8, P<0.05) . Conclusion: In the process of manned deep-sea diving, when the oceanauts perform manipulator operation tasks and troubleshooting tasks, with the increase of task difficulty, the mental load of oceanauts increases, and the blood pressure index increases significantly in a short time. At the same time, improving the proficiency of operation can reduce the variation range of blood pressure indexes. Blood pressure can be used as an effective reference to evaluate the difficulty of operation and guide scientific training.

Prevalence and influencing factors of elevated blood pressure among primary and middle school students in Anhui Province / 中国学校卫生

Objective@#To analyze the prevalence of elevated blood pressure among primary and middle school students in Anhui Province in 2022, as well as its influencing factors, so as to provide scientific evidence for the development of effective measures to prevent and control elevated blood pressure among children and adolescents.@*Methods@#Using data from the 2022 "Monitoring of Common Diseases and health Influencing Factors of Students" in Anhui Province, a total of 210 360 primary and middle school students aged 7 to 17 with complete information. Statistical analysis, including independent sample t tests, analysis of variance, and Chi square tests, were conducted to compare blood pressure differences and prevalence rates among students with different characteristics. Generalized linear mixed models were established to analyze the influencing factors of elevated blood pressure in children and adolescents.@*Results@#The overall prevalence of elevated blood pressure among primary and middle school students in Anhui Province was 22.3%, higher among high school students (25.6%) and junior high school students ( 23.7%) than elementary school students (20.4%) ( χ 2=528.49, P < 0.01 ); higher among obese(33.3%) and overweight students (24.9%) than that of normal students (18.6%) ( χ 2=3 704.52, P <0.01), higher among rural students (23.1%) than urban students (21.4%) ( χ 2=83.39, P <0.01), and higher in the northern part of Anhui Province (26.9%) than in the central part (21.9%) and southern part (17.6%) ( χ 2=1 777.51, P <0.01). The generalized linear mixed models revealed that girls, high school stage, overweight, obese, living in the northern part of Anhui Province, and living in rural were all positively correlated with elevated blood pressure( OR =1.11，1.25，1.47，1.45，2.24，1.21，1.67， P <0.01).@*Conclusions@#The prevalence of elevated blood pressure among children and adolescents in Anhui Province is the highest in the country. It is recommended that relevant departments promptly formulate corresponding measures to curb the increasing trend of elevated blood pressure and protect the physical and mental health of children and adolescents.

Single-copy Loss of Rho Guanine Nucleotide Exchange Factor 10 ( arhgef10) Causes Locomotor Abnormalities in Zebrafish Larvae / 生物医学与环境科学（英文）

OBJECTIVE@#To determine if ARHGEF10 has a haploinsufficient effect and provide evidence to evaluate the severity, if any, during prenatal consultation.@*METHODS@#Zebrafish was used as a model for generating mutant. The pattern of arhgef10 expression in the early stages of zebrafish development was observed using whole-mount in situ hybridization (WISH). CRISPR/Cas9 was applied to generate a zebrafish model with a single-copy or homozygous arhgef10 deletion. Activity and light/dark tests were performed in arhgef10 -/-, arhgef10 +/-, and wild-type zebrafish larvae. ARHGEF10 was knocked down using small interferon RNA (siRNA) in the SH-SY5Y cell line, and cell proliferation and apoptosis were determined using the CCK-8 assay and Annexin V/PI staining, respectively.@*RESULTS@#WISH showed that during zebrafish embryonic development arhgef10 was expressed in the midbrain and hindbrain at 36-72 h post-fertilization (hpf) and in the hemopoietic system at 36-48 hpf. The zebrafish larvae with single-copy and homozygous arhgef10 deletions had lower exercise capacity and poorer responses to environmental changes compared to wild-type zebrafish larvae. Moreover, arhgef10 -/- zebrafish had more severe symptoms than arhgef10 +/- zebrafish. Knockdown of ARHGEF10 in human neuroblastoma cells led to decreased cell proliferation and increased cell apoptosis.@*CONCLUSION@#Based on our findings, ARHGEF10 appeared to have a haploinsufficiency effect.

Effect of Shaoyaotang on Expressions of CD14, FADD and Caspase-8 in Colonic Tissues of Rats with Large Intestinal Damp-heat Syndrome of Ulcerative Colitis / 中国实验方剂学杂志

Objective:To observe the effect of Shaoyaotang on the contents of cell adhesion molecule-1 （ICAM-1） and transforming growth factor-<italic>β</italic>₁ （TGF-<italic>β</italic>₁） in serum of large intestine damp-heat syndrome of ulcerative colitis （UC） in rats， and the gene and protein expressions of leukocyte differentiation antigen14 （CD14）， Fas-related death domain protein （FADD） and cysteinyl aspartate specific protease-8 （Caspase-8） in the focal colon tissue. Method:A total of 80 SPF Wistar rats were randomly divided into the blank group （<italic>n</italic>=10） and modeling group （<italic>n</italic>=70）. The large intestine damp-heat syndrome of UC rats was replicated by the combination of disease and syndrome， which was high-fat， high-sugar and spicy diets combined with 2， 4-dinitrobenzene sulfonic acid （DNBS） and ethanol. After successful modeling， the modeled groups were divided into model group， sulfasalazine （SASP）control group， and low， medium and high-dose Shaoyaotang groups by the method of random number table， with14 rats in each group. Low， medium and high doses of Sulfasalazine 0.2 g·kg^-1·d^-1 and Shaoyaotang （6， 12， 24 g·kg^-1·d^-1）were given by gavage. The blank group and the model group were given equal volume of normal saline for 21 days. The contents of serum ICAM-1 and TGF-<italic>β</italic>₁ were detected by enzyme-linked immunosorbent assay （ELISA）， the expressions of CD14， FADD and Caspase-8 mRNA in colon tissues were detected by Real-time quantitative polymerase chain reaction （Real-time PCR）， and the expressions of CD14， FADD and Caspase-8 protein in colon tissues were detected by Western blot. Result:Compared with the blank group， the serum ICAM-1 level in the model group were significantly increased， whereas the content of TGF-<italic>β</italic>₁ were significantly decreased （<italic>P</italic><0.05）. The relative expression levels of CD14， FADD， Caspase-8 mRNA and protein were significantly increased （<italic>P</italic><0.05）. Compared with the model group， the content of ICAM-1 in the serum of the rats in the medium， high-dose Shaoyaotang groups and the SASP group were significantly decreased， while the content of TGF-<italic>β</italic>₁ in the serum of the rats in the low， medium， high-dose Shaoyaotang groups and the SASP group were significantly increased （<italic>P</italic><0.05）. The expression levels of CD14， FADD， Caspase-8 mRNA and protein in each intervention group were significantly decreased （<italic>P</italic><0.05）， especially in the high-dose Shaoyaotang group and the SASP group. Conclusion:Shaoyaotang has a certain intervention effect on UC rats with large intestine damp-heat syndrome， and its mechanism may be related to the inhibition of CD14， FADD and Caspase-8 genes and proteins expression.

KCNQ Channels in the Mesolimbic Reward Circuit Regulate Nociception in Chronic Pain in Mice / 神经科学通报·英文版

Mesocorticolimbic dopaminergic (DA) neurons have been implicated in regulating nociception in chronic pain, yet the mechanisms are barely understood. Here, we found that chronic constructive injury (CCI) in mice increased the firing activity and decreased the KCNQ channel-mediated M-currents in ventral tegmental area (VTA) DA neurons projecting to the nucleus accumbens (NAc). Chemogenetic inhibition of the VTA-to-NAc DA neurons alleviated CCI-induced thermal nociception. Opposite changes in the firing activity and M-currents were recorded in VTA DA neurons projecting to the medial prefrontal cortex (mPFC) but did not affect nociception. In addition, intra-VTA injection of retigabine, a KCNQ opener, while reversing the changes of the VTA-to-NAc DA neurons, alleviated CCI-induced nociception, and this was abolished by injecting exogenous BDNF into the NAc. Taken together, these findings highlight a vital role of KCNQ channel-mediated modulation of mesolimbic DA activity in regulating thermal nociception in the chronic pain state.

Effects of simulated 100 m Trimix conventional diving on oxidative stress and inflammatory reaction in rabbits / 中国应用生理学杂志

OBJECTIVE@#To investigate the effects of simulated 100 m Trimix conventional diving on tissue inflammatory cytokines in rabbits.@*METHODS@#Eight New Zealand rabbits were performed a simulated 100 m Trimix conventional diving program which was established according to the Haldane theory. The expression levels of interferon-gamma(IFN-), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8), myeloperoxidase(MPO) and matrix metallo proteinase-9 (MMP-9) in rabbits lung and brain tissues were detected by Elisa after diving decompression. The tissue wet/dry ratio was calculated. The serum levels of superoxide dismutase (SOD),glutathione(GSH), catalase(CAT), malondiadehyde(MDA) and lipid peroxide(LPO) were detected by Elisa method in rabbits before and after diving.@*RESULTS@#The expressions of IFN-, TNF-α, IL-6, IL-8, MPO and MMP-9 in simulated diving group rabbits were significantly increased compared with the intact group(<0.05, <0.01); the simulated diving rabbits tissues wet/dry ratio had no significant changes compared with the intact group. After diving, the activities of SOD and GSH were decreased significantly (<0.01), while the contents of CAT, MDA and LPO were increased significantly (<0.01).@*CONCLUSIONS@#The simulated 100 m Trimix conventional diving had significant impact on oxidative stress and inflammatory reaction in rabbits, the results of wet/dry ratio showed that the diving rabbits had no tissue edema after decompression.

Research progress of endometrial regeneration and stem cell transplantation in the treatment of intrauterine adhesions / 医学研究生学报

Despite the sharply development of reproductive science,the treatment for infertility has arrived at its bottle neck. Being a common cause of preganancy failure,intrauterine adhension(IUA)is getting widespread attention.A new treatment for IUA has brought into being when the stem cell theory of endometrial regeneration was presented.This paper summarizes the research progress of both treatment and etiology of IUA.

Beclin1, LC3 and mTOR expression in esophageal squamous cell canceration and its clinical significance / 临床与实验病理学杂志

Purpose To investigate the difference of expression of autophagy-related gene (Beclin1,LC3,mTOR) in the development of esophageal squamous cell cancer.Methods Immunohistochemical EnVision method was adopted to detect the expression of autophagy-related gene Beclinl,LC3 and mTOR in 30 cases of normal esophageal mucosa,32 cases of low-grade intraepithelial neoplasia (LGIN),34 cases of highgrade intraepithelial neoplasia (HGIN),35 cases of early carcinoma and 126 cases of advanced esophageal carcinoma,respectively.The correlation between their expression with clinicopathologic factors was also analysed.Results The expression of Beclin1 in advanced esophageal carcinoma was obviously higher than that in another four groups (P ＜ 0.005).LC3 expression in advanced esophageal carcinoma was significantly higher than that in normal esophageal mucosa,LGIN and early carcinoma (P ＜ 0.005).The expression of mTOR in advanced esophageal carcinoma was significantly higher than that in normal esophageal mucosa,LGIN and HGIN (P ＜ 0.005).In advanced esophageal carcinoma group,the expression of Beclin1,LC3 and mTOR was related to tumor TNM stage and lymph node metastasis (P ＜ 0.05).Beclin1 expression was positively associated with LC3 and mTOR expression in advanced squamous cell carcinoma (P ＜ 0.05).Positive correlation was also observed between the expression of mTOR and LC3 in advanced esophageal carcinoma and HGIN (P ＜ 0.05).Conclusion In the carcinogenesis and development of esophageal cancer,Beclin1,as a tumor suppressor gene,activates autophagy and leads to excessive self consumption and death of tumor cells.mTOR promotes tumor growth by inhibiting autophagy and promoting angiogenesis.The combined detection of Beclinl,LC3 and mTOR may be beneficial to evaluate the progression and prognosis of esophageal squamous cell cancer.

Effect of leech on VSMCs in early atherosclerosis rats via p38MAPK signaling pathway / 中国中药杂志

To explore the effect of leech on proliferation and apoptosis of vascular smooth muscle cells(VSMCs) in early atherosclerosis rats via p38MAPK signaling pathway and investigate its possible mechanism. Biochemical analyzer was used to examine the regulation of leech on levels of triglycerides(TG), total cholesterol(TC), low-density lipoprotein(LDL-C), and high-density lipoprotein(HDL-C) in blood lipid of rats. The expression of transforming growth factor-beta 1(TGF-β1) in serum was detected by ELISA. Immunological histological chemistry (IHC) was taken to measure the expression levels of proliferating cell nucleus antigen(PCNA) and cell apoptosis proteinase-3(Caspase-3), while the protein expression levels of MKK3, p38 and C-myc were detected by Western blot. In addition, hematoxylin and eosin(HE) staining was used to observe the morphological change of thoracic aortas. The results showed that leech decreased the levels of TC, LDL-C obviously and increased HDL-C, suppressed the expression levels of TGF-β1 and PCNA, up-regulated Caspase-3, down-regulated the expression levels of MKK3, p38, and C-myc protein. HE staining indicated that it could inhibit intimal thickening and reduce plaque formation. The above results indicated that leech may affect the protein expression of the p38MAPK signaling pathway to inhibit proliferation and promote the apoptosis of VSMCs via reducing blood lipid levels and suppressing TGF-β1, aiming at inhibiting intimal thickening and reducing plaque formation, tand then slowing down the process of early atherosclerosis.

Clinical Value ofF-FDG PET/CT for Patients with B Cell Lymphoma-Associated Hemophagocytic Syndrome / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the clinical value ofF-FDG PET/CT for patients with B cell lymphoma-associated hemophagocytic syndrome.</p><p><b>METHODS</b>The clinical characteristics, laboratory parameters andF-FDG PET/CT data of 23 newly diagnosed patients sufferred from B cell lymphoma-associated hemophagocytic syndrome were retrospectively analyzed. The correlation between PET and laboratory parameters were determined using Spearman correlation test. The prognostic factors were analyzed by the Kaplan-Meier method.</p><p><b>RESULTS</b>23 patients were all examined byF-FDG PET/CT before chemotherapy, theF-FDG uptake of spleen positively correlated with neutrophil count and hemoglobin content (r=0.588, P=0.035;r=0.699, P=0.008), respectively, and theF-FDG uptake of bone marrow positively correlated with neutrophil count only (r=0.691, P=0.009). Among all the clinical or laboratorial parameters andF-FDG PET/CT, only PET parameter was poor factor affecting prognosis of patients with B cell lymphoma-associated hemophagocytic syndrome. Out of 6 patients received PET/CT scans after 6 cycles of treatment, the 5 patients with negative PET/CT survived, and one patient with positive result died.</p><p><b>CONCLUSION</b>BaselineF-FDG PET/CT may provide prognostic information for the management of patients with B cell lymphoma-associated hemophagocytic syndrome, and the data ofF-FDG PET/CT before chemotherapy may predict the pregnosis of the patients with negative results, and the negative PET/CT results after chemotherapy betokens a better prognosis of patients.</p>

Calculation of underwater decompression schedule for the simulated 100 m Trimix conventional diving and verification of the schedule with animal experiment / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the underwater decompression schedule for 100 m Trimix conventional diving operations and evaluate its safety through a simulated rabbits Trimix conventional diving.</p><p><b>METHODS</b>According to the Haldane theory, the assumed time units, the classification of tissue compartments, the nitrogen super-saturation safety coefficient and the selection of methods used for the calculation of the simulated 100 m Trimix conventional diving schedule were properly selected, and the calculating method for the dive decompression schedule was thus firmly established. In our experiments, five tissue compartments were selected during the calculation of decompression schedule: 5 min, 10 min, 20 min, 40 min and 75 min, and the nitrogen super-saturation safety coefficient was calculated by 1.6. Eight New Zealand rabbits were performed a simulated 100 m Trimix dive program which was established according to the Haldane theory, and eight rabbits for intact group. The tissues wet/dry ratio and ethology were detected and observed before and after the simulated diving to evaluate the safety of decompression schedule.</p><p><b>RESULTS</b>By using the developed underwater decompression schedule, abnormal ethology changes in rabbits could not be observed after compression and decompression to the surface; and the tissues wet/dry ratio of simulated diving rabbits had no significant changes compared with the intact group (P > 0.05).</p><p><b>CONCLUSION</b>The decompression schedule calculated by Haldane theory seemed to be safe and reliable, the diving breathing gas concentration did not cause oxygen toxicity and nitrogen narcosis among the dive rabbits, and dive efficiency was greatly improved by using enriched oxygen gas in UPTD safety range during decompression.</p>

Regulation effect of adipose-derived mesenchymal stem cells on pancreatic cancer cells / 中华外科杂志

<p><b>OBJECTIVE</b>To analyze the effects of adipose tissue-derived stem cells (ADSCs) on the proliferation and invasion of pancreatic cancer (PaCa) cells and the the possible mechanism involved.</p><p><b>METHODS</b>ADSCs were isolated and co-cultured with PaCa cells. CCK-8 assay was used to detect the proliferation of PaCa cells. An ELISA was used to determine the concentration of stromal cell-derived factor-1 (SDF-1) in the supernatants. The proliferation of PaCa cells by SDF-1 was measured. AMD3100 regulated the co-culture of ADSCs and PaCa. The tumor growth of PaCa cells was assessed after treatment by ADSCs in vivo.</p><p><b>RESULTS</b>ADSCs can promote the proliferation and invasion of PaCa cells (proliferation: SW1990: 1.535 ± 0.153; PANC-1: 1.370 ± 0.100; the value of control was 1; invasion: SW1990: 47.0 ± 2.6 vs. 28.3 ± 1.3; PANC-1: 40.3 ± 1.8 vs. 24.3 ± 1.3; t = 4.332-9.558, P < 0.05). The expression of SDF-1 was high in ADSCs, but not in PaCa cells (69 ± 5 vs. 0 and 0, F = 389.134, P < 0.01). The promotion of SDF-1 on PaCa cells depends on the concentration. AMD3100 significantly downregulates these growth-promoting effects of ADSCs on PaCa cells. ADSCs significantly promoted the growth of SW1990 in nude mice at the 5(th) week (volume: (1295 ± 102) mm(3) vs. (967 ± 81) mm(3), t = 5.614, P < 0.05) , but not in PANC-1 cell.</p><p><b>CONCLUSION</b>ADSCs can promote the proliferation and invasion of PaCa cells, which may involve the SDF-1/CXCR4 axis.</p>