BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs)differentiating into neural cells is an effective way of cell therapy of nervous system disease.However,the methods used nowadays still need to be improved.OBJECTIVE:To induce the differentiation of rhesus monkey BMSCs into neuron-like cells by using sonic hedgehog factor.METHODS:Rhesus monkey BMSCs differentiating into neuron-like cells was induced by typical retinoic acid and sonic hedgehog factor.Rhesus monkey BMSCs were isolated and cultured by density gradient centrifugation method.Cell growth was observed under an inverted phase contrast microscope and cell growth curve was determined using MTT assay.Flow cytometry was performed to characterize the phenotype of BMSCs,and immunohistochemistry was utilized to assess differentiated cells.Ultra-structure of the differentiated cells was observed by transmission and scanning electron microscopes.RESULTS AND CONCLUSION:Rhesus monkey BMSCs cultured in vitro were identified by flow cytometry,with high homogenicity.Following sonic hedgehog factor disposal for 7 days,differentiated cells were mainly positive for neurone specific enolase,neurofilament protein,Tau and glial fibrillary acidic protein(GFAP).Image statistical analysis found that in sonic hedgehog factor scheme,neural stem cells marker Neetin positive rate was significantly higher compared with the rstinoic acid scheme(P＜0.01).GFAP-positive rate was greater in the retinoic acid scheme than in the sonic hedgehog factor scheme(P＜0.05).Results indicated that sonic hedgehog factor scheme is an effective pathway of rhesus monkey BMSC differentiation into neuron-like cells.

BACKGROUND: Present studies have demonstrated that during neural development, differentiation of neural stem cells (NSCs) was affected by various regulatory factors from surrounding microenvironment. Sonic hedgehog (Shh) is a key induction signal during neural fetal development, and can be an effective inductor to regulate differentiation of neural cells. OBJECTIVE: To investigate the signal transduction pathway of SHH for differentiation of rhesus monkey bone marrow mesenchymal stem cells (BMSCs) into of neuron-like cells by sonic hedgehog factor. METHODS: Rhesus monkey BMSCs were isolated and cultured by conventional density gradient centrifugation. BMSCs in the induction group were treated with L-DMEM containing FGF2, B27 and fetal bovine serum for preinduction of 24 hours, and then with DMEM supplemented with 0.5 μmol/L retinoic acid or 400 μmol/L SHH for 8 days. Non-induced cells served as control group. Following labeling with neuron enolase, positive cells were screened by flow cytometry. RT-PCR and Western-blot were used to detect SHH- and retinoic acid-induced cell membrane receptor and intracellular signal protein changes. RESULTS AND CONCLUSION: SHH specific membrane receptor Ptc, retinoic acid specific receptor RARα, signal protein molecule ptch1 and Smad expressed in normal cells. Ptc expression upregulated in SHH-induced cells. High expression lasted for a long time with induction time, which was significantly stronger compared with the retinoic acid and control groups (P < 0.01). Intracellular ptch1 protein molecule expression showed similar tendency as this, but could not induce upregulation of RARα expression. During induction, retinoic acid-stimulated cells did not activate Ptc pathway. Four days following induction, RARα expression upregulated and lasted till 6 days, but there were no significant differences. No significant change in ptch1 expression was determined. SHH- and retinoic acid-induced cell Smad molecule expression upregulated, but no significant difference was determined. Results verified that SHH-induced scheme participated in cell induction and differentiation by persistently activating its specific receptors. However, there was no significant receptor pathway crossing between retinoic acid-induced and SHH-induced schemes.

Artemisinin and its derivatives have shown the potential application value in anti-tumor fields.But their applications are limited due to poor solubility, short half-life and so on;therefore, many scholars devoted themselves to developing and studying relative pharmaceutical preparations.Nano drug delivery system is a new drug delivery tool.Compared with the traditional dosage forms, such as tablets, suppositories and injections, nano drug delivery system has such advantages as good stability, targeted drug release and combined drug delivery.In this review, nano drug delivery carriers for artemisinin were summarized, including nanoparticles, liposomes, micelles, nanomicroemulsions and so on.The research results and characteristics of artemisinin-containing nano formulas in tumor therapy, as well as the co-delivery system of artemisinin and transferrin, were discussed.

Background and purpose:Δ133p53 can promote tumor cell growth, but the exact mechanism is not clear. This study was aimed to observe the expression and signiifcant of the p53 isoformsΔ133p53 and p53 gene downstream molecules MDM2 and cyclin G1 genes by 5-FU-MKN45 gastric cancer cell line model. Methods:Af-ter using different concentrations of 5-FU (50μg/mL, 100μg/mL) to human gastric cancer cell line MKN45, inhibition rate should be detected by MTT assay, the changes ofΔ133p53 mRNA, MDM2 mRNA and cyclin G1 mRNA express-ing were detected by reverse transcription-polymerase chain reaction (RT-PCR). Differences between these groups were analyzed by ANOVA, comparisons within groups were analyzed by t-test, bivariate correlation was analyzed by Pearson linear correlation. Results:MTT results showed that with the increased concentration of 5-FU and the extension of time, the cell inhibition rates increased gradually. The inhibition rates of 50μg/mL 5-FU were 41.10%, 54.79%and 68.48%, for culturing 24, 48 and 72 hours. There were statistically signiifcant differences between the groups(F=45.52, P=0.00). The inhibition rates of 100μg/mL 5-FU were 69.53%, 78.21%and 86.92%, for culturing 24, 48 and 72 hours. There were statistically signiifcant differences between the groups(F=85.58,P=0.00). The inhibition rates of 50 and 100μg/mL were 41.10%and 69.53%, for culturing 24 hours. There were statistically signiifcant differences between the groups(F=51.29, P=0.00). The inhibition rates of 50 and 100μg/mL were 54.79%and 78.21%, for culturing 48 hours. There were statistically signiifcant differences between the groups(F=51.29, P=0.00). The inhibition rates of 50 and 100μg/mL were 68.48%and 86.82%, for culturing 72 hours. There were statistically signiifcant differences between the groups(104.91, P=0.00). RT-PCR results showed that with the increase of the concentration of 5-FU, theΔ133p53 mRNA, MDM2 mRNA and cyclin G1 mRNA expression gradually declined in gastric cancer cell line MKN45 cells, and there were statistically signiifcant differences between the groups(F=738.532, 1 396.607, 2 785.56,P=0.00). Cor-relation analysis showed that the expressions ofΔ133p53 mRNA and MDM2 mRNA in gastric cancer were positively correlated (r=0.871, P=0.01), while the expression of cyclin G1 mRNA and p53 mRNA had no obvious relevance (P=0.13). Conclusion:In the 5-FU-MKN45 gastric cancer cell line model, anti-tumor pathway ofΔ133p53 isomers is related with MDM2 but was not related with cyclin G1.

ObjectiveTo investigate the role of p38 and ERK1/2 during rhesus monkeys mesenchymal stem cells differentiated into neuron-like cells.MethodsTo induce the neuronal phenotype,rhesus monkeys mesenchymal stem cells were maintained in sub-confluent cultures in serum-contain medium supplement with Sonic hedgehog.Western blot analysis the change of p38 and ERK1/2 during rhesus monkeys mesenchymal stem cells differentiated into neuron-like cells.Under transmission and scanning electron microscope,ultra-structure of the differentiated cell were observed.ResultsDuring BMSCs differentiated into neuron-like cells by SHH,Mitogen-activated protein kinases (MAPK) involved in their signal transduction,p38 was activated and ERK1/2 was inhibited.P38 inhibitor SB203580 increased induced differentiation time compared with normal induced cells,and inhibited neurite outgrowth.ConclusionActivation of p38 and inhibition of ERK was impacted on differentiation into neuron-like cells from rhesus monkeys mesenchymal stem cells induced by Sonic hedgehog,which may has potential application on neuroprotection of stem cells in Nervous system diseases

Objective To assess the safety and efficacy of percutaneous nephrolithotripsy withpneumatic and ultrasonic power in pediatric renal calculi. Methods The clinical data of 44 patientstreated with combination of pneumatic and ultrasonic power during pereutaneous nephrolithotripsywere retrospectively analyzed. The 44 patients had 49 renal calculi. The patients were all under 14years old. The average age was 11 years (range 7-14 years). There were 39 unilateral and 5 bilateralcalculi. Among the 44 patients,metabolic disturbance occurred in 19 cases (43.2%),anatomical dys function occurred in 15 cases (34.1%),urinary tract infection occurred in 14 cases (31.8%). Ante grade percutaneous access was established under ultrasound guidance,a combination of pneumatic andultrasonic lithotripsy were used. The effect was evaluated by postoperative KUB and ultrasonic. Re suits The access was successfully established in all patients. Complete stone clearance was achievedin 36 kidneys in phase Ⅰ,stones from 9 kidneys were completely removed with second lithotripsy.Leftover stone in 2 kidneys were treated by ESWL. Open surgery was performed in 2 kidneys due toexcessive bleeding. The operative time ranged from 52 132 min,average time was 79 min. Two pa tients needed blood transfusion. No severe complications occurred in all patients. Thirty seven pa tients were followed up for 3 18 months. The renal function was not worsened and hydronephrosiswas not aggravated in these patients. Conclusion The percutaneous nephrolithotripsy with pneumatic and ultrasonic power is a safe,effective treatment for pediatric renal calculi.

Neurofibromatosis type 2 is an autosomal dominant inherited disease with a low incidence. It often involves the central and peripheral nervous systems, leading to bilateral hearing loss, balance difficulties, facial paralysis and other symptoms, which seriously affects the quality of life of patients. Therefore, in the treatment of neurofibromatosis type 2, preservation of facial nerve function is crucial. The diagnosis and treatment scheme should take into account the characteristics of the tumor and the patient′s wishes. Individualized programmes of multidisciplinary collaboration are needed.

Objective To detect serum oxytocin and highly sensitive C-reactive protein (hs-CRP) levels in obese and type 2 diabetes mellitus(T2DM) subjects and investigate the relationships between serum oxytocin levels and hs-CRP, glycolipid metabolism, insulin resistance and pancreas β cell function. Methods A total of 176 subjects were enrolled in the study, including 88 patients with newly-diagnosed type 2 diabetes ( T2DM) and 88 subjects with normal glucose tolerance(NGT). NGT and T2DM groups were further divided each into normal weight (NW) and obese(OB) subgroups. Obesity was defined as body mass index(BMI)≥25 kg/ m2 according to the WHO-Western Pacific Region diagnostic criteria (2000). 75g oral glucose tolerance test ( OGTT) was performed in all subjects. Fasting plasma glucose ( FPG), 2 h postprandial plasma glucose (2hPG), fasting insulin ( FINS), 2h postprandial serum insulin(2hINS), HbA1C and lipids were also determined. Insulin resistance and pancreas β-cell function were determined by homeostasis model assessment ( HOMA-IR, HOMA-β). Highly sensitive C-reactive protein(hs-CRP) level was determined by chemiluminescence immunoassay and fasting serum oxytocin level was determined by ELISA. Results Serum oxytocin level was lower in T2DM group than that in NGT group(P<0. 01), while serum hs-CRP level was higher in T2DM group than that in NGT group(P<0. 01). The level of serum oxytocin in subjects with obesity was also lower than that in subjects with NW in both NGT and T2DM groups [7. 16(6. 45-8. 82) vs 7. 98(7. 03-9. 17) ng/ L and 9. 23(8. 16-10. 36) vs 9. 86(8. 77-12. 06) ng/ L, P<0. 05]. The level of serum hs-CRP in subjects with obesity was higher than that in subjects with NW in both NGT and T2DM groups [0. 99(0. 25-1. 97) vs 0. 54(0. 19-0. 91) mg/ L and 3. 47(1. 63-6. 20) vs 1. 65(0. 81-3. 81) mg/ L, P<0. 05]. Serum oxytocin level was negatively correlated with hs-CRP, BMI, WC, WHR, HbA1C , FPG, 2hPG, FINS, 2hINS, total cholesterol, triglycerides, LDL-C and HOMA-IR, while was positively correlated with HOMA-β(P<0. 05). Subjects within the upper serum hs-CRP tertile had lower level of oxytocin when compared to subjects in the middle or lower serum hs-CRP tertiles(P<0. 05 ). Conclusion Serum oxytocin level was decreased in subjects with type 2 diabetes as well as with obesity. Serum oxytocin level was closely correlated with inflammation, glycolipid metabolism, insulin resistance, and pancreas β cell function. It may play an important role in the pathogenesis of obesity and T2DM.

Objective:Primary biliary cholangitis (PBC) and Primary Sj?gren′s syndrome (pSS) are autoimmune epithelial inflammatory diseases that share many common clinical symptoms. The aim of this study was to investigate the differences and diagnostic value of Autotaxin (ATX) in PBC and SS.Methods:The clinical data of 237 cases diagnosed with PBC, PBC secondary to SS, pSS and healthy individuals(HC) between September 2020 and September 2021 were retrospectively analyzed. The levels of ATX in each group were measured by enzyme-linked immunosorbent assay (ELISA), and the corresponding sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and area under the curve ( AUC), etc were analyzed. Normally distributed data were expressed as mean ±SD and non-normally distributed as median (IQR). The differences and correlations between ATX and the biochemical tests in each group were assessed by applying the Mann-Whitney U test, Spearman correlation analysis, etc. P<0.05 was considered statistically significant difference. Results:The results showed that ATX was positive in 33.9%, 33.3% and 53.3% for PBC, PBC secondary SS, and pSS, respectively, with the specificities of 93.1%, 100% and 93.2%, respectively. The highest accuracy was achieved in pSS and the sensitivity and specificity were 86.5% and 93.2%, which were higher than those in PBC group(56.8%, 93.1%), respectively. Compared with HC [32.6(21.8, 60.5)ng/ml], ATX levels in PBC[59.3(48.6, 86.3)ng/ml, U=1 750.50, P<0.001], PBC-SS [73.6 (53.3,102.4)ng/ml; U=199.00, P<0.001], and pSS [152.6 (97.4,192.1)ng/ml, U=264.00, P<0.001] were elevated with significant difference ( P<0.05). ATX levels showed a decreasing trend from the pSS group to the HC group. ATX in PBC group[AUC(95% CI)= 0.73(0.651,0.812), P<0.001], PBC secondary SS group [AUC(95% CI)=0.82(0.730, 0.912), P<0.001], and pSS group [AUC(95% CI)=0.94(0.898, 0.984), P<0.001] had prediction accuracy. ATX was associated with total protein ( r=-0.31, P=0.041) level and glutaminase (r=-0.26, P=0.024) level. Conclusion:ATX has diagnostic value in both PBC and SS, and with higher sensitivity and specificity for the latter.

Background Prolonged exposure to vibration can cause vascular endothelial injury, and inflammatory response plays an important role in vascular endothelial injury. Studies have shown that long non-coding RNA (lncRNA) maternally expressed gene 3 (MEG3) is involved in regulating the expression of inflammatory injury of endothelial cells. Objective To investigate the effects of vibration on the secretion of inflammatory factors and the expression of IncRNA MEG3 by vascular endothelial cells in vitro. Methods Human umbilical vein endothelial cells (HUVEC) were divided into two categories: vibration and control. The vibration exposure included 63 Hz (6.76 m·s⁻²), 200 Hz (5.08 m·s⁻²), and 250 Hz (4.56 m·s⁻²) frequency bands, and 1 and 2 d exposure time with 1 to 4 h of daily vibration. The control treatment was the same as the vibration category except that they were not exposed to vibration. CCK-8 was used to detect the effects of different vibration frequencies and time on the viability of HUVEC. The expression levels of tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8), interleukin-4 (IL-4), and interleukin-10 (IL-10) in the cells and supernatants were detected by enzyme-linked immunosorbent assay. The expression levels of IncRNA MEG3 were detected by real-time fluorescence quantitative PCR. Results Compared with the cells with the control treatment, the cell viability of the 1-day exposure group increased after 1.5 h and 3 h of vibration at 63 Hz, while decreased after 2 h and 2.5 h; the cell viability of the 2-day exposure group increased at the frequency of 63 Hz for 1.5 h, but decreased at 2 h and 2.5 h. At the frequency of 200 Hz, the cell viability of the 1-day exposure group increased at 2 h and 4 h, but decreased at 2.5 h and 3 h; the cell viability of the 2-day exposure group increased at 1.5 h and decreased at 2.5 h. For the vibration exposure at frequency of 250 Hz, the cell viability of the 1-day exposure group increased at 1.5 h and 2.5 h, but decreased at 3 h; of the 2-day exposure group, the cell viability increased at 1.5 h and decreased at 3 h. For the exposure settings of 63 and 200 Hz vibration for 2.5 h and 250 Hz vibration for 3 h, and with the control treatment as reference, the expression levels of TNF-α, IL-8, IL-4, and IL-10 in cells and supernatants were increased in the 1 d and 2 d exposures; the expression level of lncRNA MEG3 decreased in the 1 d exposure group; however, for the 2 d exposure, the expression level of lncRNA MEG3 decreased only in the 63 Hz vibration exposure. All of these results were statistically significant (P<0.05). Conclusion Vibration could induce an increase in the levels of inflammatory factors TNF-α, IL-8, IL-4, and IL-10 and a decrease in the expression level of lncRNA MEG3 in vascular endothelial cells in vitro.