Objective:To explore the involvement of miR-126 and the role of mammalian target of rapamycin (mTOR)/hypoxia-induced factor 1 α (HIF-1 α) pathway in regulating human umbilical cord mesenchymal stem cells (hUCMSCs) exosomes (Exo) on vascular endothelial growth factor (VEGF)-A levels in high glucose-induced human retinal vascular endothelial cells (HRECs).Methods:The hREC was cultured in EGM-2-MV endothelial cell culture medium with 30 mmol/L glucose and placed in hypoxic cell incubator by 1% oxygen concentration. The cell model of high glucose and low oxygen was established. After modeling, divided HRECs into Exo group, phosphate buffer saline (PBS) group, PBS+anti-miR126 group, Exo+anti-miR126 group, PBS+anti-mTOR group, and PBS+anti-HIF-1 α group. High-glucose and hypoxia-induced hREC in the PBS and Exo groups were respectively co-cultured with PBS and 100 μg/ml hUCMSC Exo. PBS+anti-mTOR group, PBS+anti-HIF-1 α group: 500 nmol/L mTOR inhibitor ADZ2014, 25 μmol/L HIF-1 α inhibitor YC-1 pretreatment for hREC 12 h, and then co-culture with PBS after High-glucose and hypoxia-induced. PBS+anti-miR126 group, Exo+anti-miR126 group: miR-126 LNA power inhibitor probe was transfected with high glucose, and co-cultured with PBS and hUCMSC Exo 6 h after transfection. Real-time polymerase chain reaction (qPCR) measured miRNA-126 expression levels in PBS, and Exo groups for 0, 8, 16 and 24 h. After 24 h of co-culture, the levels of mTOR and HIF-1 α in the cells of PBS and Exo groups were detected by immunofluorescence, Western blot and qPCR, respectively. Western blot, qPCR detection of VEGF-A expression levels in cells of the PBS+anti-mTOR and PBS+anti-HIF-1 α groups. The expression of VE GF-A, mTOR, and HIF-1 α mRNA was measured in cells of PBS+anti-miR126 group and Exo+anti-miR126 group by qPCR. Comparison between two groups was performed by t-test; one-way ANOVA was used for comparison between multiple groups. Results:At 0, 8, 16 and 24 h, the relative mRNA expression of miR-126 gradually increased in the Exo group ( F=95.900, P<0.05). Compared with the PBS group, The mTOR, HIF-1 α protein ( t=3.466, 6.804), mRNA in HRECs in the Exo group, VEGF-A mRNA expression ( t=8.642, 7.897, 6.099) were all downregulated, the difference was statistically significant ( P<0.05). The relative expression level of VEGF-A protein ( t=3.337, 7.380) and mRNA ( t=8.515, 10.400) was decreased in HRECs of the anti-mTOR+PBS group and anti-HIF-1 α+PBS group, differences were statistically significant ( P<0.05). The relative expression of VEGF-A, mTOR, and HIF-1 α mRNA was significantly increased in the cells of the Exo+anti-miR126 group, the differences were all statistically significant ( t=4.664, 6.136, 6.247; P<0.05). Conclusions:miR-126 plays a role in regulating the effect of hUCMSCs exosomes on VEGF-A levels in high glucose-induced HRECs via mTOR-HIF-1 α pathway.

【Objective】 To explore the relationship between the storage time of leukodepleted red blood cells and transfusion adverse reactions by analyzing the occurrence of transfusion adverse reactions of patients after leukodepleted red blood cells transfusion from four hospitals. 【Methods】 By using the electronic medical record management system, the collection and transfusion dates of leukodepleted red blood cells from four hospitals in Enshi Prefecture from 2018 to 2022, as well as the information on transfusion adverse reactions, were retrieved. 【Results】 From 2018 to 2022, a total of 697 61 bags of leukodepleted red blood cells were transfused in four hospitals, resulting in 166 cases of transfusion adverse reactions, among which 93 were allergic reactions, 63 were non hemolytic febrile reactions, and 10 were others, with a total incidence rate of transfusion adverse reactions at 0.24%. The average storage time of leukodepleted red blood cells with and without transfusion adverse reactions was (20.25±6.31) and (19.88±5.50) days, respectively. With a storage time of 7 days as the threshold, the incidence of transfusion adverse reactions was the lowest for a storage time of 15~21 days. The incidence of transfusion adverse reactions of leukodepleted red blood cells in two groups (with storage days ≤21 days and >21 days) was not statistically significant(P>0.05). 【Conclusion】 Allergic reactions were the main type of transfusion adverse reaction caused by leukodepleted red blood cells, and the incidence of transfusion adverse reactions decreased and then increased with the prolongation of the storage time of leukodepleted red blood cells. There was no significant difference in the incidence of transfusion adverse reactions with leukodepleted red blood cells stored for ≤ 21 days and >21 days.

Objective:To explore the risk factors of paralytic ileus (PI) after simultaneous pancreas-kidney (SPK) transplantation.Methods:From January 2017 to December 2019, clinical data were reviewed retrospectively for 115 cases of SPK transplantation. The risk factors of PI after SPK were analyzed. According to the occurrence of PI, they were divided into two groups of occurrence and non-occurrence. One-way analysis of variance was utilized for analyzing such influencing factors as gender, age, body mass index (BMI), diabetic type, duration of diabetes, mode of dialysis, duration of dialysis, diabetic gastroenterology, history of open surgery, bowel preparation, operative duration, hemorrhagic volume, immunosuppressant and hypoproteinemia. Multivariate Logistic regression analysis was performed for screening the suspected risk factors.Results:Among them, 19 patients (16.5%) had PI. Univariate analysis showed that PI was associated with diabetic gastroenterology, operative duration, history of open surgery, no bowel preparation and hypoproteinemia ( P<0.05). Multivariate Logistic regression analysis revealed that the risk factors of PI after SPK included diabetic gastroenterology, operative duration time, history of open surgery and no bowel preparation ( P<0.05). Conclusions:Diabetic gastroenterology, operative duration, history of open surgery and no bowel preparation are risk factors for PI after SPK. Clinical interventions for the above factors are necessary.

After brachial plexus avulsion (BPA), microglia induce inflammation, initiating and maintaining neuropathic pain. EZH2 (enhancer of zeste homolog 2) has been implicated in inflammation and neuropathic pain, but the mechanisms by which it regulates neuropathic pain remain unclear. Here, we found that EZH2 levels were markedly upregulated during BPA-induced neuropathic pain in vivo and in vitro, stimulating pro-inflammatory cytokines (IL-1β, TNF-α, and IL-6) secretion in vivo. In rats with BPA-induced neuropathic pain, mechanical and cold hypersensitivities were induced by EZH2 upregulation and inhibited by EZH2 downregulation in the anterior cingulate cortex. Microglial autophagy was also significantly inhibited, with EZH2 inhibition activating autophagy and reducing neuroinflammation in vivo. However, this effect was impaired by inhibiting autophagy with 3-methyladenine, suggesting that the MTOR signaling pathway is a functional target of EZH2. These data suggest that EZH2 regulates neuroinflammation and neuropathic pain via a novel MTOR-mediated autophagy signaling pathway, providing a promising approach for managing neuropathic pain.

To summarize the clinical experience regarding a patient with early recurrence of atypical hemolytic uremic syndrome (aHUS) after renal transplantation. AHUS is a rare disease with high recurrence rate and poor prognosis. Although the patient was treated with plasma exchange, intravenous gamma globulin, rituximab block B lymphocyte, hormone shock and so on, he still suffered renal transplantation failure. The risk of aHUS recurrence after renal transplantation should be fully evaluated.

Objective: The purpose of this study is to investigate the expression change of cell cycle-related molecules in platal tissue of fetal mice with cleft palate, induced by 2, 3, 7, 8-Tetrachlorodibenzo-p-dioxin (TCDD), and to explore the mechanism of cell cycle-related molecules in cleft palate. Methods: In vivo, 48 pregnant mice were randomly divided into TCDD treatment group and control group with Random number table, 24 mice in each group. On the embryonic day 10.5 (E10.5), pregnant mice were orally administrated with TCDD 28 μg/kg (containing 5 μg/ml TCDD of corn oil) in TCDD treatment group. The same volume of corn oil was given to the mice in control group. The pregnant mice in each group were sacrificed on E13.5, E14.5 and E15.5, to collect the fetal palates for analysis. Fetal palates were used to extract total RNA and total protein, so as to detect the expression levels of cell cycle-related molecules, using RT-PCR and western blotting respectively. In vitro, human kidney embryo 293t (HEK293t) cells were treated with different concentrations of TCDD (0.01, 0.1, 0.5 and 1 nmol/L), and cells proliferation activity was detected using MTT assay. Statistical analysis was performed with IBM SPSS 24.0. Kolmogorov-Smimov test was used for normal distribution check, and the distribution was normal. Independent t-test was carried out among two groups. P<0.05 was considered statistically significant. Results: At E13.5, E14.5 and E15.5, the expression level of interferon regulatory factor 6 (Irf6) protein were higher in the control group (1.26 ± 0.13, 1.67 ± 0.14 and 1.42 ± 0.15, respectively) compared to that in the TCDD group (0.81 ± 0.08, 1.04± 0.02 and 0.86 ± 0.12, respectively), on each time point (t value were 2.836, 3.662 and 2.867, respectively; P values were 0.0471, 0.0146 and 0.0241, respectively). The expression level of cyclin-dependent kinase inhibitor 1A (P21) protein on E13.5 and E14.5 of the control group (2.26 ± 0.21, 1.99 ± 0.21)were higher than that in the TCDD group on each time point(1.43 ± 0.12、0.93 ± 0.22), (t value were 3.398 and 3.378; P value were 0.8726 and 0.0273). The expression level of cyclin D1 in the control group (1.00±0.02, 0.94±0.03 and 1.11±0.09, respectively)were higher than that of the TCDD group (0.28±0.01, 0.33±0.06 and 0.88±0.01, respectively) on each time point (t value are were 22.53, 22.35 and 14.27, respectively, P value <0.001, <0.001 and<0.001, respectively). The expression of cyclin E1, cyclin A2, cyclin B1, CDK6, CDK2 and CDK1 in TCDD groups were higher than that of the controls (P<0.05). However, there was no significant difference of cyclin B1 on E13.5 and Cdk2 on E15.5. As treatment with TCDD (0.1 nmol/L) at 1, 2 and 3 days (0.70 ± 0.05, 1.05 ± 0.03 and 1.39 ± 0.04, respectively), the proliferation of HEK293t cells increased compared with the control group (0.49 ± 0.04, 0.98 ± 0.03 and 1.55 ± 0.02, respectively). The differences were statistically significant (t value were 2.829, 1.395 and 2.692, respectively; P value were 0.0198, 0.1320 and 0.0247, respectively). Conclusions TCDD down-regulates Irf6 and P21, and interferes with the normal expression of cell cycle-associated molecules, which in turn interferes with medial edge epithelia (MEE) cells cycle arrest and proliferation. These indicate that the disorder of spatiotemporal expression of cell cycle-related molecules during palatal development may be involved with the mechanism of TCDD-induced cleft palate..

Objective: To evaluate the expression of histone H4 acetylation(Ac-H4) during the cleft palates formation induced by 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin(TCDD) in C57BL/6J mice. Methods: Forty-eight pregnant C57BL/6J mice were completely randomly divided into two groups: ① TCDD group, mice were treated with 20ug/kg of TCDD on gestation day (GD) 10.5 by gastric perfusion; ② control group, mice were treated with an equivalent of corn oil. The head samples were collected and sliced in coronal plane on GD13.5, GD14.5 and GD15.5 respectively. Histone H4 acetylation in the palates were evaluated by immunohistochemical staining and Western Blot in the two groups. Results: Histone H4 acetylation was mainly expressed in the palatal epithelial cells and slightly expressed in mesenchymal cells. The expression level of histone H4 acetylation was 0.6002±0.2530, 0.9180±0.0941 and 0.8966±0.0908 respectively in control group on GD13.5, GD14.5 and GD15.5; while 1.0229±0.2779, 1.6095±0.2651 and 1.2758±0.1251 in TCDD group. There were statistically significant differences between the control group and TCDD group (P<0.05). Conclusions The histone H4 acetylation was involved in the cleft palate formation induced by TCDD in C57BL/6J mice.

Objective: To study the anti-inflammatory effect and mechanism of Gutong plasters in acute inflammatory model of rats. Methods:Totally 60 SD rats were randomly divided into the blank group,the model group,Gutong plasters at low,medium and high dose groups(0.594,1.188 and 2.376 g/patch containing crude drug 0.48,0.96 and 1.92 g,respectively) and prednisone ace-tate group(0.005 4 g·kg-1). The acute inflammation model was prepared by injecting 5% formaldehyde into right side of foot plan-tar. And then,the anti-inflammatory effect was evaluated by measuring the foot plantar volume. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of nitric oxide(NO),histamine(HIS) and 5-hydroxytryptamine(5-HT) in serum and inflam-matory tissue,the levels of prostaglandin E2(PGE2),tumor necrosis factor alpha(TNF-α),interleukin 1 beta(IL-1β) and interleu-kin 6 (IL-6) in inflammatory tissue were also determined. Pathological changes were observed through the pathological sections pre-pared by HE staining. Results:When compared with the model group,Gutong plasters could significantly inhibit the swelling of foot plantar in inflammatory rats(P<0.01). At the same time,Gutong plasters could significantly reduce the levels of NO,5-HT and HIS in serum and inflammatory tissue in different degrees(P<0.05 or P<0.01),significantly reduce the content of PGE2in inflammatory tissue (P<0.05 or P<0.01),increase the content of IL-6 in inflammatory tissue and improve the pathological changes of inflammato-ry tissues (the pathological score was significantly reduced). In partical indictor changes,Gutong plasters and prednisone acetate showed an equal effect.In addition,Gutong plasters didn't show significant dose-dependent manner in inhibiting foot swelling,affect-ing inflammatory mediators and improving pathological changes(P>0.05). Conclusion:Gutong plasters have better anti-inflammato-ry effect in the acute inflammation model induced by formaldehyde in rats. The anti-inflammatory effect may be associated with the de-crease of the levels of NO,5-HT and HIS in serum and inflammatory tissue,the decrease of the content of PGE2and the increase of the content of IL-6 in inflammatory tissue. In addition,the anti-inflammatory effect may be relevant to improving the local inflammatory tis-sue subcutaneous edema and inflammatory cell infiltration.

Objective To study the expression and significance of Glypican-3 in Budd-Chiari syndrome (BCS) complicated with hepatocellular carcinoma (HCC).Methods The data of 46 patients with BCS complicated with HCC (the BCS + HCC group) treated in The First Affiliated Hospital of Zhengzhou University from January 2007 to December 2016 were analyzed retrospectively.Another 48 patients with HBV-related HCC (the HBV + HCC group) and 43 patients with hepatic cyst (the hepatic cyst group) were randomly selected as the control groups during the same time period.The differencesin positive rates of Glypican-3 in the liver tissues among the three groups were compared.The BCS + HCC group was further divided into the Glypican-3 positive and Glypican-3 negative subgroups according to the expression of Glypican-3.The differences in gender,age,AFP,HbsAg,Child-Pugh classification,tumor number,extrahepatic metastasis,vascular invasion,Edmondson-Steiner grading and BCLC staging between the two subgroups were compared.The survival time of the two subgroups was compared using the Kaplan-Meier method.Results The expression rates of Glypican-3 in the BCS + HCC group,HBV + HCC group and Hepatic Cyst group were 76.1％,70.8％ and 0％,respectively.The levels of Glypican-3 in the BCS + HCC group and the HCC group were significantly higher than that in the hepatic cyst group.The differences were statistically significant (P ＜ 0.05).No statistically significant difference was detected between the BCS + HCC group and the HBV + HCC group (P ＞ 0.05).In the group of patients with BCS + HCC,there was no significant difference in gender,age,AFP,HbsAg,Child-Pugh classification,tumor number and extrahepatic metastasis between the Glypican-3 positive and negative subgroups (P ＞0.05).However,vascular invasion,Edmondson-Steiner grading and BCLC staging in the Glypican-3 positive subgroup were significantly higher than those in the Glypican-3 negative group,(P ＜ 0.05).The 1-year,3-year and 5-year survival rates were 77.1％,51.0％ and 22.8％ in the Glypican-3 positive subgroup,compared with 90.9％,63.6％ and 45.5％ in the Glypican-3 negative subgroup,respectively.There were statistically significant differences between the two groups (P ＜ 0.05).Conclusion Glypican-3 has a stable expression in patients with BCS complicated with HCC,and it is closely related to malignancy of the tumor and prognosis of the patients.