Objective:To explore the mechanism of peritoneal dialysis solution (PDS)-induced peritoneal microinflammation through activation of ceramide (CER) in peritoneal dialysis model mice.Methods:Thirty 5-week-old male C57BL/6 mice weighing about 22 g were used to set up peritoneal dialysis models, and then were randomly divided into 4 groups: sham operation group (1.5 ml sterilized water, n=7), high glucose-PDS group (1.5 ml 4.25% PDS, n=8), high glucose-PDS+ acid sphingomyelinase (ASMase) inhibitor desipramine (DES) group (1.5 ml sterilized water+10 mg/kg DES, n=8), high glucose-PDS+Src kinase inhibitor PP2 group (1.5 ml sterilized water +1 mg/kg PP2, n=7), with intraperitoneal injection once a day. After 28 days, the mice were sacrificed to retain peritoneal tissues. HE staining and Masson staining were used to observe the histological changes of peritoneum. Immunohistochemistry was used to detect the Toll-like receptor 4 (TLR4) and macrophages. High performance liquid chromatography, liquid chromatography/mass spectrometry and immunofluorescence were used to detect the expression of ASMase and CER. Real-time quantitative PCR was used to detect the mRNA levels of c-Src, p-Src, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Western blotting was used to detect the protein levels of c-Src, and p-Src. Enzyme-linked immunosorbent assay was used to detect the serum C reactive protein (CRP), IL-6 and TNF-α. Results:(1) High glucose-PDS led to peritoneal hyperplasia, collagen deposition and fibrosis in the peritoneal dialysis mice, indicating successful modeling. Compared with high glucose-PDS group, peritoneal hyperplasia, collagen deposition and fibrosis of mice treated with DES and PP2 were significantly improved (all P<0.05). (2) Compared with sham operation group, ASMase activation and CER level of peritoneal tissues were significantly higher in high glucose-PDS group, and DES could significantly inhibit activated ASMase and increased CER expression caused by high glucose-PDS (both P<0.05). PP2 had no significant effect on ASMase activation and CER level (both P>0.05). (3) Compared with sham operation group, there were more TLR4 and macrophage positive staining cells in peritoneal tissues in high glucose-PDS group, and the mRNA expression levels of IL-6 and TNF-α in peritoneal tissues and serum CRP, IL-6 and TNF-α were higher (all P<0.05). DES and PP2 could significantly inhibit the increased TLR4, macrophages and related inflammatory factors induced by high glucose-PDS (all P<0.05). (4) Compared with sham operation group, c-Src and p-Src mRNA and protein expression levels of peritoneal tissues in high glucose-PDS group were significantly higher (all P<0.05). PP2 significantly inhibited the increased p-Src mRNA and protein levels caused by high glucose-PDS (both P<0.05), but had no significant effect on the mRNA and protein expression levels of c-Src (both P>0.05). DES had no significant effect on the mRNA and protein expression levels of c-Src and p-Src (all P>0.05). Conclusions:High glucose-PDS may enhance the expression of CER through stimulating the activity of ASMase, phosphorylate Src, activate TLR4 and induce inflammatory damage of peritoneum in peritoneal dialysis model mice.

Venous thromboembolism (VTE) is a complication in children with acute lymphoblastic leukemia (ALL). The Chinese Children's Cancer Group-ALL-2015 protocol was carried out in China, and epidemiology, clinical characteristics, and risk factors associated with VTE were analyzed. We collected data on VTE in a multi-institutional clinical study of 7640 patients with ALL diagnosed in 20 hospitals from January 2015 to December 2019. First, VTE occurred in 159 (2.08%) patients, including 90 (56.6%) during induction therapy and 108 (67.92%) in the upper extremities. T-ALL had a 1.74-fold increased risk of VTE (95% CI 1.08-2.8, P = 0.022). Septicemia, as an adverse event of ALL treatment, can significantly promote the occurrence of VTE (P < 0.001). Catheter-related thrombosis (CRT) accounted for 75.47% (n = 120); and, symptomatic VTE, 58.49% (n = 93), which was more common in patients aged 12-18 years (P = 0.023), non-CRT patients (P < 0.001), or patients with cerebral thrombosis (P < 0.001). Of the patients with VTE treated with anticoagulation therapy (n = 147), 4.08% (n = 6) had bleeding. The VTE recurrence rate was 5.03% (n = 8). Patients with VTE treated by non-ultrasound-guided venous cannulation (P = 0.02), with residual thrombus (P = 0.006), or with short anticoagulation period (P = 0.026) had high recurrence rates. Thus, preventing repeated venous puncture and appropriately prolonged anticoagulation time can reduce the risk of VTE recurrence.
Humans ; Child ; Venous Thromboembolism/etiology* ; East Asian People ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/epidemiology* ; Risk Factors ; Thrombosis/chemically induced* ; China/epidemiology* ; Anticoagulants/adverse effects* ; Recurrence

Objective To explore the regulatory effect of miR-26b-5p on PI3K/PIP3 signaling path-way and its effect on L-type calcium channel of atrial cardiomyocytes in rats with ischemic ar-rhythmia.Methods A total of 72 SD rats were randomly divided into sham operation group,mod-el group,LY294002 group,negative control group,overexpression group and combination group,with 12 rats in each group.In 24 h after corresponding intervention,rat model of ischemic arrhyth-mia was established in the other groups except the sham operation group(only thoracotomy but without ligation).During the modeling process,arrhythmia indexes were detected in each group,and Curtis-Walker scoring was used for arrhythmia score.Fluorescence quantitative PCR was em-ployed to detect the expression of miR-26b-5p in atrial myocardiocytes,whole-cell patch-clamp technique was used to record the ICa-L of atrial myocardiocytes,and Western blotting was applied to measure the expression of CACNA1C,calmodulin and PI3K/PIP3 pathway related proteins.Re-sults Compared with the model group,the number of left ventricular preventricular contrac-tions,duration of ventricular tachycardia,duration of ventricular fibrillation,arrhythmia score,ab-solute value of ICa-L peak density of atrial myocardiocytes,and expression levels of CACNA1C and calmodulin were significantly increased,and the levels of miR-26b-5p,p-PI3K,PIP3 and p-Akt were obviously decreased in the LY294002 group(P＜0.05).While the overexpression group had decreased number of left ventricular preventricular contractions,shorter duration of ventricular tachycardia,shorter duration of ventricular fibrillation,lower arrhythmia score,reduced absolute value of ICa-L peak density,and lower expression levels of CACNA1C and calmodulin,and in-creased levels of miR-26b-5p,p-PI3K,PIP3 and p-Akt than the model group(P＜0.05).The number of left ventricular preventricular contractions,duration of ventricular tachycardia,dura-tion of ventricular fibrillation,arrhythmia score,absolute value of ICa-L peak density and expres-sion levels of CACNA1C and calmodulin were significantly increased,and the expression of p-PI3K,PIP3 and p-Akt were obviously decreased(0.82±0.08 vs 1.09±0.11,0.91±0.09 vs 1.17± 0.11,0.94±0.09 vs 1.20±0.12,P＜0.05)in the combination group than the overexpression group.Conclusion Overexpression of miR-26b-5p may block the L-type calcium channel in atrial myocardiocytes in ischemic arrhythmia rats by activating PI3K/PIP3 related signaling pathway,and thus improve the performance of arrhythmia in rats.

Objective: To evaluate the antiseptic effect of combined using of 5% sodium hypochlorite and calcium silicate-based root canal sealer against Enterococcus faecalis (Ef) biofilms in infected dentinal tubules in vitro. Methods: Cells of Ef were inoculated into the dentinal tubules of single-rooted teeth (without caries, periapical lesions and malformations extracted due to periodontal disease or orthodontic reasons; collected from Department of Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University) with centrifugation and incubated in brain-heart infusion (BHI) to form 3-week-old biofilms. The infected samples were subjected to sodium hypochlorite or sterile water bathing for 10 minutes followed by calcium silicate-based root canal sealer (iRoot SP) (calcium silicate-based group), Gutta-percha group and sterile water group placed on the root canal wall for 1, 4 and 12 weeks. There were two samples in each treatment at each point. The antiseptic effectiveness of combined use of sodium hypochlorite and calcium silicate-based root canal sealer was analyzed by laser scanning confocal microscope (LSCM), ANOVA and LSD-t test. Results: After treatment with 5% sodium hypochlorite, in calcium silicate-based group for 4 and 12 weeks more Ef biofilm cells [(75.3±3.5)% and (74.8±3.8)%] were killed than in Gutta-percha group [(65.9±4.1)% and (63.0±3.7)%] and sterile water group [(63.9±4.0)% and (64.2±3.5)%] (P<0.05). After being treated with sterile water, the proportion of dead bacterial cells in calcium silicate-based group for 1, 4 and 12 weeks [(27.5±4.6)%, (43.0±4.4)% and (40.3±6.1)%] were more than those in Gutta-percha group and sterile water group (P<0.05). After being treated with 5% sodium hypochlorite or sterile water, more biofilm bacteria were killed in calcium silicate-based group for 4 and 12 weeks than in calcium silicate-based group for 1 week (P<0.05). Conclusions The combined use of sodium hypochlorite and calcium silicate-based root canal sealer kills more biofilm cells in infected dentinal tubules.

To evaluate the antiseptic effect of combined using of 5% sodium hypochlorite and calcium silicate?based root canal sealer against Enterococcus faecalis (Ef) biofilms in infected dentinal tubules in vitro . Methods Cells of Ef were inoculated into the dentinal tubules of single?rooted teeth (without caries, periapical lesions and malformations extracted due to periodontal disease or orthodontic reasons; collected from Department of Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University) with centrifugation and incubated in brain?heart infusion (BHI) to form 3?week?old biofilms. The infected samples were subjected to sodium hypochlorite or sterile water bathing for 10 minutes followed by calcium silicate?based root canal sealer (iRoot SP) (calcium silicate?based group), Gutta?percha group and sterile water group placed on the root canal wall for 1, 4 and 12 weeks. There were two samples in each treatment at each point. The antiseptic effectiveness of combined use of sodium hypochlorite and calcium silicate?based root canal sealer was analyzed by laser scanning confocal microscope (LSCM), ANOVA and LSD?t test. Results After treatment with 5% sodium hypochlorite,in calcium silicate?based group for 4 and 12 weeks more Ef biofilm cells [(75.3 ± 3.5)% and (74.8 ± 3.8)% ] were killed than in Gutta?percha group [(65.9±4.1)% and (63.0±3.7)%] and sterile water group [(63.9±4.0)% and (64.2±3.5)%] (P<0.05). After being treated with sterile water, the proportion of dead bacterial cells in calcium silicate?based group for 1, 4 and 12 weeks [(27.5±4.6)%, (43.0±4.4)% and (40.3±6.1)%] were more than those in Gutta?percha group and sterile water group (P<0.05). After being treated with 5% sodium hypochlorite or sterile water, more biofilm bacteria were killed in calcium silicate?based group for 4 and 12 weeks than in calcium silicate?based group for 1 week (P<0.05). Conclusions The combined use of sodium hypochlorite and calcium silicate?based root canal sealer kills more biofilm cells in infected dentinal tubules.

In May 2015, a child with absence of most of the five fingers with scar formation after healing of a left hand burn wound hospitalized in our burn ward. According to the free online design program for making artificial limbs using three-dimensional printing technology on the internet, a utility artificial hand, most of which made of plastic parts, was designed for the child and printed by a three-dimensional printer. The child was instructed to wear and use the utility artificial hand, including driving the finger part of the utility artificial hand to make a grasping action by flexing the wrist joint. On the first day of using the utility artificial hand, the time the right hand and the utility artificial hand took to finish the Nine-Hole Peg Test (NHPT) was 24 and 325 s, respectively. After training, the child could grab some light and rough objects. After 3 months of follow-up, the child could use the utility artificial hand to cooperate with the upper limb of the healthy side to make the movements of picking up the basketball and keeping the balance of body on the bicycle. The time the right hand and the utility artificial hand took to finish NHPT was 21 and 193 s, respectively. The time the utility artificial hand took increased by 40.6% compared with the initial period. By assembling the three-dimensionally printed utility artificial hand, the partial appearance image of the child was restored, and some of the hand functions were compensated, which improved the self-care ability of the child in daily life and was beneficial to his physical and mental development.

Objective: To evaluate the antimicrobial activity of nonequilibrium plasma against Enterococcus faecalis (Ef) biofilms in vitro and to obtain novel evidence of root canal disinfection with nonequilibrium plasma. Methods: Sterile cover slips and single-rooted canals were filled with Ef and incubated to form 1-week-old and 3-week-old biofilms, respectively. The infected samples were subjected to nonequilibrium plasma, 2% chlorhexidine digluconate (CHX) and saline for 3, 10 and 30 minutes, respectively. After treatment, the killing effectiveness of nonequilibrium plasma was analyzed by using laser scanning confocal microscopy (LSCM) and colony forming unit (CFU) counting. Results: The 3-dimentional reconstruction LSCM images showed that about 48.3%-79.8% of 1-week-old Ef biofilm cells and 40.0%-67.4% of 3-week-old biofilm cells were killed by nonequilibrium plasma and 2% CHX compared to saline (P<0.05). The proportion of killing activity was lower after 3 minutes (40.0%-50.9% killing) than after 10 minutes (65.3%-77.8% killing) and 30 minutes (66.4%-79.8% killing) (P<0.05). And the killing of biofilm bacteria was fastest during the first 3 minutes (13.3%-17.0% killing per minute) and slow down greatly after 10 minutes. Remarkably more bacteria were killed in 1-week-old Ef biofilms (48.3%-79.8% killing) than in 3-week-old biofilms (P<0.05). Conclusions The nonequilibrium plasma killed more Ef biofilm cells in infected root canals showed promotional as an additional approach against bacterial biofilms during root canal disinfection.

Objective: To observe the clinical effects of the Joint Active System on the treatment of joint dysfunction after deep burn. Methods: Twenty-two patients with joint dysfunction after deep burn were hospitalized in Institute of Burns of Tongren Hospital of Wuhan University & Wuhan Third Hospital from January 2015 to October 2016, involving 18 elbow joints with flexion disorder, 10 wrist joints with dorsal extension disorder, and 12 ankle joints with dorsal extension disorder. They were treated with the elbow joint activity training device, the wrist joint activity training device, and the ankle joint activity training device of the Joint Active System, respectively. The treatment was carried out 3 times each day with interval of 6 h, 30 minutes each time, and it lasted for four to seven months, with one month as a course of treatment. Before treatment and 1, 2, 3, 4 month (s) after, active motion range of each joint was measured by joint goniometer. Function improvement of each joint was evaluated, and the total effective ratio was calculated 4 months after treatment. Satisfaction degree of patients was assessed by the modified Likert Scale 1, 2, 3, 4 month (s) after treatment. Data were processed with one-way analysis of variance for repeated measurement and LSD test. Results: Before treatment and 1, 2, 3, 4 month (s) after, flexion active motion range of elbow joints were (61±23), (78±22), (89±20), (96±20), and (103±19)°; dorsal extension active motion range of wrist joints were (23±7), (31±6), (38±9), (44±5), and (49±8)°; dorsal extension active motion range of ankle joints were (-31±12), (-23±10), (-16±7), (-12±6), and (-8±4)°, respectively. The active motion range of each joint was obviously higher 1, 2, 3, 4 month (s) after treatment than the previous time point of the same joint (with P values below 0.01). Four months after treatment, the total effective ratios of function improvement of elbow joints, wrist joints, and ankle joints were 5/6, 9/10, and 2/3, respectively. Scores of satisfaction degree of the patients 1, 2, 3, 4 month (s) after treatment were (1.3±0.7), (2.2±1.0), (2.8±0.8), and (3.3±0.6) points, respectively. Scores of satisfaction degree of the patients were obviously higher 2, 3, 4 months after treatment than the previous time point (with P values below 0.05). Conclusions Joint Active System can improve the active range of motion of each joint obviously in treating joint dysfunction after deep burn, with total effective ratio of function improvement of each joint surpassing 0.66, and the majority of patients are quite satisfied with the curative effects.

ObjectiveToexploretheeffectof ceremideonprocess of peritoneal mesothelial cells(PMCs) apoptosis induced by peritoneal dialysis solution(PDS).Methods PMCs were cultured with normal DMEM,1.5％ PDS and 4.25％ PDS.4.25％ mannitol was used as high osmotic pressure control.Ceremide were detected by LC-MS-MS.Flow cytometry was used in apoptosis analysis.Bax,p53 and bcl-2 protein expressions were detected by Western blotting.Results (1) PDS caused the increase of intracellular ceremide in PMCs,and normal and high osmotic pressure controls had no such effect.As the acidic sphigomyelinase inhibitor,desipramine significantly inhibited the production of ceramide induced by 4.25％ PDS [(56.08±12.24) μg/L vs (91.25:t:15.89) μg/L,P＜0.01]. (2) Compared with 1.5％ PDS,4.25％ PDS stimulated PMCs apoptosis (26.65％±6.21％ vs 4.04％±1.86％,P＜0.01),up-regulated bax and p53 proteins expression (P＜0.01),and down-regulated bcl-2 protein exprssion(P＜0.05).Desipramine obviously inhibited the apoptosis induced by 4.25％ PDS,decreased bax and p53 proteins expression,increased bcl-2 protein expression(P＜0.05).Exogenous C2-ceremide reversed the effect of desipramine(P＜0.05).Conclusion The increase of intracellular ceremide may play an important role in the PMCs apoptosis induced by high glucose PDS.

OBJECTIVE: To investigate the mechanism of the protective effect of Cordyceps sinensis (C. sinensis) on the apoptosis of cultured NRK-52E induced by angiotension II (AngII). METHODS: NRK-52E cells were incubated with C. sinensis (0, 5, 10, 20, and 40 mg/L) and 10(-8) mol/ L AngII for 24, 48, 72 h. The optimal concentration of C. sinensis was selected. Either NRK-52E cells were incubated with different doses of AngII (0, 10(-12), 10(-10), 10(-8), and 10(-6) mol/L) for 24 h, or with 10(-8) mol/L AngII for 24, 48, and 72 h, to observe the effect of AngII on the apoptosis of NRK- 52E cells. The optimal concentration and time of AngII were selected. In another experiment cells were divided into 5 groups: a control, AngII (10(-8) mol/L), AngII (10(-8) mol/L)+ C. sinensis (40 mg/ L), Ang II (10(-8) mol/L)+ fosinopril (10(-5) mmol/L), and Ang II (10(-8) mol/L)+ fosinopril (10(-5) mol/ L)+C. sinensis (40 mg/L). MTT assay was used to test the changes in the proliferation of NRK-52E cultured with different concentration of C. sinensis for 24, 48, 72 h. The Annecxin V-FITC and PI stainings were applied to detect the apoptosis rate induced by AngII by flow cytometer (FCM) and to determine the eddects of C. sinensis. The activity of caspase-3 was assayed by spectrophotometry. RESULTS: Certain concentrations of C. sinensis (10-40 mg/L) promoted the proliferation of NRK- 52E cells inhibited by AngII(P<0.05). AngII induced the apoptosis of NRK-52E in a dose and timedependent manner, accompanied with increased activity of caspase-3 (P<0.05). C. sinensis partially suppressed the apoptosis of NRK-52E induced by AngII, and declined the activity of caspase-3 (P<0.05). No significant difference was shown as between the fosinopril group and the fosinopril+C. sinensis group (P>0.05). CONCLUSION C. sinensis can suppress the apoptosis of NRK-52E by AngII, and the protective effect of C. sinensis may be inhibiting the activation of caspase-3 during the AngII-induced apoptosis of NRK-52E.
Angiotensin II ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line ; Cells, Cultured ; Cordyceps ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; cytology ; Humans ; Kidney Tubules ; cytology ; Protective Agents ; pharmacology