1.Iodine effect on survival time of neuron-like cells differentiated from bone marrow mesenchymal stem cells
Wenzhu YAN ; Shujian QIN ; Xuezheng LIU ; Dehua LI
Chinese Journal of Tissue Engineering Research 2010;14(10):1716-1720
BACKGROUND:In vitro experiment has shown that the survival time of conventional chemical induction-induced neuron-like cells differentiated from bone marrow mesenchymal stem cells(BMSCs)was short,which limited its further application.With regard to the possibility of extension of iodine-induced neuron-like cells,the survival time has not yet been professionally reported.OBJECTIVE:To research the effects of the micro-element iodine on the survival time of neuron-like cells differentiated by BMSCs.METHODS:Rat mesenchymal stem cells at passage 3 were obtained under sterile condition,and divided into groups A-F.In group A,iodine ion was not added.In groups B-F,iodine ion at mass concentrations of 2,55,90,125 and 2 500 mg/L was added respectively.An additional blank control group was established,and simultaneously the cells were induced into neuron-like cells with dimethyl sulphoxide(DMSO).Cells following induction were subjected to immunohistochemistry.Survival time of neuron-like cells was observed under different mass concentrations of iodine ion.RESULTS AND CONCLUSION:When mass concentrations of iodine ion were between 55-125 mg/L,the survival time of neuron-like cells prolonged to about 5 days and structures of induced cells were intact.From then on,the number of dead cells was gradually increased till approximately one week,all neuron-like cells died.When mass concentrations of iodine ion were 2 mg/L and 2 500 mg/L,cell survival time was from 12-36 hours.No significant difference was determined compared with group A.Till 2 or 3 days,all neuron-like cells died.Above-described results indicated that an appropriated concentration of iodine iron added in the common chemical induction may be benefit for the survival time of the neuron-like cells differentiated by BMSCs,but the effect may be negligible for the survival time of neuron-like cells induced when the added concentration of iodine iron is too low or too high.
2.Differentiation of rat bone marrow mesenchymal stem cells into neuron-like cells in vitro:Induction of Wnt3a signaling molecules
Wenzhu YAN ; Shujian QIN ; Xuezheng LIU ; Dehua LI
Chinese Journal of Tissue Engineering Research 2010;14(14):2476-2480
BACKGROUND:Wnt signaling pathway is the key to regulation of cell proliferation and differentiation.The evidence suggests that this pathway participates in the neural precursor cell proliferation,differentiation and determination of the regulation of cell fate.At present,the Wnt signaling pathway effects on the mesenchymal stem cells(MSCs)differentiated into neuron-like cells have not been reported.OBJECTIVE:To seek Wnt signal molecule that promotes the differentiation of MSCs into neuron-like cells.METHODS:MSCs were isolated from the femur of Sprague Dawley rats in vitro using the density gradient centrifugation,and then cultured.Following passage,cell surface markers CD29,CD44,CD34 and CD45 were measured using morphology and flow cytometry.These cells were selected and evaluated.Using neurotrophic factor and basic fibroblast growth factor combined with Wnt3a and Wnt5a induction scheme,effects of Wnt3a and Wnt5a during the differentiation of MSCs into neuron-like cells were compared utilizing immunohistochemistry and reverse transcription-polymerase chain reaction(RT-PCR).Simple culture of basic fibroblast growth factor served as controls.RESULTS AND CONCLUSION:Following culture and passage of MSCs,cells adhered to the wall,showing even spindle shape.Flow cytometry showed great expression of CD29 and CD44 and low expression of CD34 and CD45.Following Wnt3a induction,cells were positive for nestin,neuron specific enolase,but no significantly expressed glial fibrillary acidic protein.Following induction,cell viability was good.In the Wnt5a induction and control groups,weakly positive expression of nestin was found,but neuron specific enolase and glial fibrillary acidic protein were negative.RT-PCR results demonstrated that nestin expression was detected in the Wnt3a induction group before and after induction.Significant amplified bands for neuron specific enolase were detected at day 5 following induction,and became more significant at day 10.Weak bands for glial fibrillary acidic protein were visible at day 10 following induction.These indicated that Wnt3a can promote the differentiation of MSCs into neuron-like cells in vitro.
3.Expression and significance of p53,nm23 and p16 in Wilms' tumor of children
Jie SUN ; Yeming WU ; Guohua LIU ; Wenzhu ZHANG ; Zhongde ZHANG ;
Chinese Journal of Tissue Engineering Research 2001;5(16):154-155
We studied 22 Wilms'tumors of children immunohistochemically.We've found that the positive rate of p53 in slices was 31.8% (7),of nm23 was 50% (11),and of p16 was 86.4% (19).It suggested that mutation rate of p53 was high in tumors,expression of nm23 in favorite histology(FH)was higher than that in unfavorite histology(UFH) group,and p16 showed very high positive rate in tumors.All of the three showed no relation with sex,age,or pathological type.So each one may be useful in clinic to evaluate pathogenesis and prognosis.
4.Clinical efficacy analysis of dorsal root ganglion pulse radio frequency stimulation combined with prega-balin in treatment of postherpetic neuralgia
Wenzhu WANG ; Ran WANG ; Jian LIU ; Sheng YANG
The Journal of Clinical Anesthesiology 2014;(7):663-665
Objective To observe the clinical curative effect of thoracic dorsal root ganglion pulse radio frequency stimulation combined with pregabalin in treatment of postherpetic neuralgia (PHN).Methods Sixty-two PHN patients were randomly divided into the single drug group and combination group (n=31).The pre-and post-treatment VAS values and pittsburgh sleep quality in-dex (PSQI),decreased dosage of tramadol after treatment,adverse reaction during treatment,and long-term therapeutic effects were observed.Results VAS values,PSQI and tramadol dosage de-creased significantly post-treatment in both groups,especially in the combination group (P < 0.05). In addition,the incidence of dizziness,somnolence and ataxia was less,but puncture pain was more in combination group (P < 0.05 ).Long-term therapeutic effects of patients in the combination group were significantly improved.Conclusion Thoracic dorsal root ganglion pulsed radio frequence com-bined with pregabalin is effective in the treatment of PHN.
5.A Microfluidic Chip with Integrated Microelectrode for Real-time Dopamine Detection
Junshan LIU ; Qinglong XIAO ; Dan GE ; Yangyang ZHANG ; Wenzhu ZHANG ; Zheng XU ; Chong LIU ; Liding WANG
Chinese Journal of Analytical Chemistry 2015;(7):977-982
A microfluidic chip with integrated microelectrode for real-time dopamine detection was designed and fabricated. The chip consisted of a polydimethylsiloxane ( PDMS) channel plate and a glass electrode plate. One central channel as the culture chamber of neural stem cells and two lateral channels for transport of the culture medium were integrated on the PDMS channel plate. Microelectrodes for real-time dopamine detection were integrated on the glass electrode plate. To solve the problem in demoulding the PDMS channel plate from the silicon mould, a novel demoulding method was developed. An Au-Au-Au three-electrode system was constructed, and it performed well in electrochemical detection. The performance of the microfluidic chip was primarily studied by detecting dopamine dissolved in the medium for the culture of neural stem cells. The limit of detection was 3. 92 μmol/L, the linear detection range was from 10 μmol/L to 500 μmol/L, and the detection reproducibility from different chips was less than 4%.
6.Local injection of simvastatin for repair of mild-to-moderate unstable humeral fractures:effects on bone healing and bone density
Jian LIU ; Baofang TIAN ; Wenzhu WANG ; Weidong ZHENG ; Li FENG ; Baoyin DU ; Dawei TIAN
Chinese Journal of Tissue Engineering Research 2015;(37):5933-5937
BACKGROUND:Studies have shown that a single injection of smal-dose simvastatin can significantly improve bone microstructure and promote trabecular bone remodeling of the femoral condyle after osteoporosis. OBJECTIVE:To investigate the effect of local single injection of simvastatin on the prognosis of mild-to-moderate unstable humeral fractures. METHODS:A total of 93 patients with mild-to-moderate unstable humeral fracture were selected and randomized into injection group (49 cases) and normal group (44 cases). The normal group received normal physical therapy, and the injection group underwent local single injection of simvastatin based on the physical therapy. Bone mineral density and healing time were compared between two groups. RESULTS AND CONCLUSION:There was no difference in the bone mineral density between the two groups within 2 weeks after treatment (P>0.05), but the bone mineral density was significantly higher in the injection group than the normal group at 1-2 months after treatment (P<0.05). In addition, the injection group was superior to the normal group in the fol owing aspects:incidence of complications, excel ent-good rate of joint function recovery, and hospital stay. These findings indicate that the local injection of simvastatin based on conventional physical therapy can shorten the time of fracture healing, increase bone mineral density and improve the prognosis of patients with mild-to-moderate unstable proximal humeral fractures.
7.Bilateral external fixation combined with limited internal fixation in the repair of open fractures of middle and distal tibia and fibula:reliable fixation on the broken ends
Jian LIU ; Baofang TIAN ; Wenzhu WANG ; Weidong ZHENG ; Li FENG ; Baoyin DU ; Dawei TIAN
Chinese Journal of Tissue Engineering Research 2015;(39):6350-6354
BACKGROUND:Open fracture of tibia and fibula is a common fracture of long bones. Simple external fixation is often difficult to achieve thorough and effective reduction and fixation. Shortcomings of open reduction and internal fixation highlighted in the high incidence of postoperative complications, and seriously affected the recovery of joint function. Therefore, external fixation combined with internal fixation for repair of open fractures of tibia and fibula has been extensively used in the clinic.
OBJECTIVE:To explore the repair efficacy of bilateral external fixation combined with limited internal fixation on open fractures of middle and distal tibia and fibula.
METHODS: Clinical data of 56 patients, who were identified by X-ray or CT examination and were diagnosed as having open fractures of middle and distal tibia and fibula in the Jining No.1 People’s Hospital, as the treatment group, were retrospectively analyzed from January 2009 to January 2013. Patients were subjected to thorough debridement, reduction of the fracture fragments, limited internal fixation and fixed bilateral external fixation within 8 hours. They received stage I suture. When the local conditions of the wound permitted, they received stage II grafting, and were repaired with adjacent muscle flap or free flap. The efficacy and postoperative complication were observed. Above data were compared with those of 44 patients with open fractures of middle and distal tibia and fibula who received reduction and internal fixation (control group).
RESULTS AND CONCLUSION:Among the 56 patients in the treatment group, the outcomes were excelent in 35 cases, good in 16 cases, average in 4 cases and poor in 1 case. The excelent and good rate was 91%. Among the 44 patients in the control group, the outcomes were excelent in 23 cases, good in 10 cases, average in 7 cases and poor in 4 cases. The excelent and good rate was 75%. The excelent and good rate was significantly higher in the treatment group than that of the control group (P < 0.05). Significant differences in incision length, operation time, fracture healing time and bone nonunion rate were detected between the two groups, and above indexes were better in the treatment group than in the control group (P < 0.05). These findings verify that bilateral external fixation combined with limited internal fixation for open fractures of middle and distal tibia and fibula is reliable, can significantly reduce postoperative complications after internal fixation, promote the healing of fracture, and is conducive to the early recovery training of the affected limb.
8.Efficacy and safety of allopurinol in the treatment of chronic kidney disease: a meta-analysis
Shunyao LIU ; Jing E ; Hongyan LUO ; Li BAO ; Wenzhu TIAN ; Xi BAO ; Shilu CAO ; Yali ZHENG
Journal of Chinese Physician 2022;24(4):505-511
Objective:To evaluate the efficacy and safety of allopurinol in the treatment of chronic kidney disease.Methods:The databases of Embase, PubMed and the Cochrane library were searched for randomized controlled trials of allopurinol in patients with chronic kidney disease. According to the Cochrane system evaluation method, two evaluators independently screened the literature and extracted the data, and analyzed the results with Revman 5.3 software.Results:Finally, 10 articles were included, including 940 patients (472 in the experimental group and 468 in the control group). Meta analysis showed that allopurinol treatment could reduce blood uric acid ( MD=-2.40, 95% CI: -2.74--2.05, P<0.01), 24-hour urinary protein ( MD=-0.61, 95% CI: -1.17--0.06, P=0.03) and increase estimation of glomerular filtration rate(eGFR) ( MD=2.51, 95% CI: 1.86-3.17, P<0.01). There was no significant difference in adverse events between the experimental group and the control group ( OR=1.40, 95% CI: 0.61-3.19, P=0.42), but allopurinol treatment could reduce the risk of cardiovascular events ( OR=0.58, 95% CI: 0.38-0.89, P=0.01). Conclusions:Allopurinol treatment of chronic kidney disease can reduce urinary protein, improve eGFR, and reduce the risk of cardiovascular events.
9.Clinical and pathological characteristics of primary IgA nephropathy patients with different blood types
Shunyao LIU ; Jing E ; Jing LI ; Jing LI ; Bo LI ; Danna MA ; Wenzhu TIAN ; Yali ZHENG
Journal of Chinese Physician 2023;25(4):516-520,527
Objective:To investigate the differences of clinical data and pathological changes in patients with primary IgA nephropathy (IgAN) with different blood types.Methods:The clinical and pathological data of patients with primary IgAN diagnosed by renal biopsy in the People's Hospital of Ningxia Hui Autonomous Region from May 2016 to May 2021 were collected. They were divided into groups A, O, B and AB according to blood group. The clinical manifestations and pathological changes of the four groups during renal biopsy were analyzed.Results:A total of 258 patients with primary IgAN were included, including 87 cases of type A, 74 cases of type O, 72 cases of type B and 25 cases of type AB. The male to female ratio was 1.34∶1, and the median age was 36 (29, 47) years old. There was no significant difference in age, sex, blood pressure, hemoglobin and renal function among the four groups (all P>0.05). Neutrophil gelatinase-associated lipocalin (NGAL) in patients with type A and B was higher than other groups (all P<0.05). There were no significant differences in mesangial cell hyperplasia (M), capillary cell hyperplasia (E), glomerular segmental sclerosis (S), renal tubule atrophy/interstitial fibrosis (T), crescent body (C) lesions and proportion of sclerosed glomeruli among the four groups (all P>0.05). Subgroup analysis by gender showed that the hemoglobin, uric acid and creatinine of male patients were higher than those of female patients (all P<0.05), but the estimated glomerular filtration rate (eGFR) and urinary protein had no statistical significance (all P>0.05). Women with blood type A and O were heavier than men under microscope. The pathological manifestations of M, E, S and C lesions in women with type A blood were heavier than those in men, and S and T lesions in men with type B blood were heavier than those in women. There was no significant difference in the general baseline data, inflammation and kidney indexes between the four groups of men and women (all P>0.05). Pathologically, the M lesions of men with B blood group were more severe than those of other blood groups, while the S and T lesions of women with B blood group were less severe than those of other blood groups. Conclusions:The clinical and pathological manifestations of IgAN women with type A are heavier, the pathological manifestations of IgAN women with type B are lighter, but the pathological lesions of IgAN men with type B are heavier.
10.Optmization of condition in oxygen-glucose deprivation/re-oxygenation models of HT22 cells
Lixi ZHAO ; Wenzhu WANG ; Tong ZHANG ; Hui CHEN ; Lixu LIU
Chinese Journal of Neuromedicine 2016;15(1):55-59
Objective To optimize the HT22 cell oxygen-glucose deprivation/re-oxygenation models in hippocampal neurons of mice,discuss the basis of model establishment and test the related parameters.Methods HT22 cell growth curve was plotted by cell counting and cell doubling time was calculated,and then,the oxygen-glucose deprivation time and re-oxygenation time were determined by LDH assay.The CCK-8 assay,cell counting,cell morphology and flow cytometry were performed to further optimize the parameters of the models.Results The HT22 cell doubling time was 17 h 18 min.The oxygen-glucose deprivation time was 18 h and the re-oxygenation was 20 h.CCK-8 assay indicated that absorbance in cells of oxygen-glucose deprivation 18 h/re-oxygenation 20 h (OGD 18 h/R20 h) group was 0.334±0.138,which was significantly lower than that in the control group (0.756±0.026,P<0.05);cells in the OGD18 b./R20 h group were shrank obviously.Flow cytometry indicated that the percentage of cells in terminal apoptosis or non-viable non-apoptotic cells was 24.56%±5.76% and that of viable apoptotic cells was 2.63%±0.55% in the OGD18 h/R20 h group,which were significantly higher than those in the control group,respectively (8.13%±0.87% and 0.80%±0.10%,/P<0.05).Conclusion The HT22 cell oxygen-glucose deprivation/re-oxygenation models are successfully established;both oxygen-glucose deprivation time and re-oxygenation time are related to the cell doubling time,and OGD18 h/R20 h can preferably help the establishment;.