BACKGROUND:Neural stem cells (NSCs) hold self-renewal and multi-directional differentiation potential. NSCs differentiation into neurons in high proportion under induction conditions exhibits broad application prospect. OBJECTIVE:To explore the effect of basic fibroblast growth factor (bFGF)-chitosan carrier on the NSCs differentiation into neurons in vitro, and whether the differentiated neurons could form synaptic-like connection with myocytes. METHODS:After purification, the NSCs were co-cultured with chitosan, soluble bFGF or bFGF-chitosan carrier. After 7-day induction, the NSCs differentiation into neurons was observed by immunofluorescence staining of beta tubulin Ⅲ. The NSCs differentiation into cholinergic neurons was observed through double immunofluorescence staining of ChaT and beta tubulin Ⅲ. The synaptic-like connection between the neurons and myocytes was observed by triple staining of beta tubulin Ⅲ and MHC. RESULTS AND CONCLUSION:The percentage of differentiated neurons in the bFGF-chitosan carrier group was 74%, which was significantly higher than that in the other two groups. Additionally, the synaptic-like connection formed between the differentiated neurons and myocytes. To conclude, the bFGF-chitosan carrier promotes the NSCs differentiation into neurons to form synaptic-like connection with the co-cultured myocytes.

Objective To compare the application of CUBIC and iDISCO clearing methods in observing 3D imaging of spinal cord with immunofluorescent staining. Methods 1 mm thick spinal cord coronal sections were processed with CUBIC and iDISCO, respectively. The neurofilament (NF) protein was detected by immunofluorescent staining and then was observed by a laser confocal microscope. Results Compared with CUBIC, iDISCO had the advantages of shorter time, higher transparency (F=6.64, P<0.01), and deeper penetration (F=5117.55, P<0.01). Conclusion Immunofluorescent staining combined with iDISCO could completely observe the spinal axons with shorter time and better stain effect.

Objective To observe the effects of neurotrophin 3 (NT3)-chitosan on motor function, and proliferation and differentiation of the neural stem cells (NSCs) in the injury area and subventricular zone (SVZ) in rats with motor cortex injury. Methods Sixty-five Wistar rats were divided into control group (n=7), injury group (n=29) and NT3-chitosan group (n=29). The motor cortex was aspirated and re-moved as cerebral injury model. NT3-chitosan was immediately implanted into the injured area after operation, and the control group re-ceived no intervention. Pellet reaching test was performed to detect the recovery of the forelimb function, HE staining was used to observe the lesion cavity size, and immunofluorescence staining was used to observe the proliferation and differentiation of NSCs 3 days, 7 days, 14 days, 1 month, 2 months and 3 months after operation. Results The grasp success rate was higher (F>6.00, P≤0.05), and the lesion cavity size was significantly smaller (F>629.5, P<0.001) in the NT3-chitosan group than in the injury group. In the NSCs differentiation experi-ment, the number of BrdU cells at all the time points was significantly higher in the NT3-chitosan group than in the injury group (F>171.43, P<0.001). In the NSCs proliferation experiment, the number of BrdU positive cells was still significantly higher in the NT3-chitosan group than in the control group and in the injury group (F>155.06, P<0.001), the number of Dcx positive cells was significantly higher in the NT3-chitosan group than in the injury group (F=62.367, P<0.001), and the number of BrdU/Dcx positive cells was significantly higher in the NT3-chitosan group than in the control group (F=33.527, P<0.001). Conclusion NT3-chitosan could activate NSCs in the SVZ, and pro-mote endogenous neurogenesis and forelimb function recovery in rats after motor cortex injury.

Objective To investigate the potential mechanism of basic fibroblast growth factor (bFGF)-chitosan carrier to induce neural stem cells to differentiate into neurons. Methods After purification, the neural stem cells were cocultured with chitosan, soluble bFGF and bFGF-chitosan carrier. Three hours, twenty-four hours, three days and seven days after induction, immunofluorescence staining of Nestin, beta tubulin III, microtubule-associated protein-2 (MAP2), and fibroblast growth factor receptor 1 (FGFR1) were used to observe the expres-sion of FGFR1;real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) and Western blotting were used to detect RNA and protein level changes after induction. Results Three hours after induction, there was no significant difference in the expression of FGFR1 among three groups. Twenty-four hours after induction, the expression level of FGFR1 was significantly higher in the bFGF-chitosan carrier group than in the chitosan group and the soluble bFGF group (P<0.001);three days and seven days after induction, the expression of FGFR1 decreased significantly in the chitosan group and soluble bFGF group (P<0.001), however, it was still higher in the bFGF-chitosan carrier group;moreover, the expression of genes associated with the pathway of extracellular regulated protein kinases/mitogen activated protein ki-nase (Erk/MAPK) was significantly higher in the bFGF-chitosan carrier group than in the chitosan group and soluble bFGF group (P<0.001). Conclusion bFGF-chitosan carrier might upregulate the expression of FGFR1, then activate Erk/MAPK signal pathways, and finally promote the differentiation of neural stem cells into neurons.

We used immobilized lipase from Candida sp. 99-125 to produce fatty acid methyl esters (FAMEs) from crude oil and methanol. We studied the effects of phospholipids on activity of immobilized lipase, reaction velocity, stability of immobilized lipase and the stability of immobilized lipase in crude and refined oil. Results showed that the activity of the lipase immersed in petroleum ether with 1% phospholipids dropped more quickly than the lipase in petroleum ether without phospholipids. When soybean oil was used without phospholipids as material, the FAMEs yield of 15 min was 26.2%, whereas the yield decreased to 12.4% when there were 5% phospholipids in the soybean oil. However when the phospholipids content was below 1%, the stability of the lipase did not change obviously. The lipase was stable when used to catalyze crude soybean oil and crude jatropha oil, after 10 cycles the FAMEs yield was still above 70%. This lipase showed great potential for industrial production of biodiesel from crude oil.
Biofuels ; analysis ; Candida ; enzymology ; metabolism ; Enzymes, Immobilized ; Fatty Acids ; metabolism ; Lipase ; metabolism ; Methanol ; metabolism ; Methyl Ethers ; metabolism ; Petroleum ; metabolism ; Phospholipids ; metabolism

Objective To evaluate the diagnostic value of ultrasound image features combined with cesarean section scores for placenta accreta . Methods A total of 151 pregnant women in the later trimester of pregnancy underwent two-dimensional and color Doppler ultrasonography . According to the results of the operation ,they were divided into the accreta group and the non-accreta group . The number of cesarean section and placental thickness in the two groups were recorded . The ultrasound image characteristics of the placenta previa ,the placenta posterior gap disappeared ,the placental lacuna in the placenta ,the muscle layer becoming thin or disappeared after the placenta ,the anatomical region of the uterus-bladder junction line and the abundant blood flow signal behind the placenta were compared between the two groups . Logistic regression analysis was performed according to the results of single factor analysis . After assigning the characteristics of the regression model ,the ROC curve was drawn to determine the best diagnostic intercept point . Results The presence of placenta previa ,placental lacuna and placental enriched blood flow signals combined with cesarean section scores were used to diagnose placenta accreta ,the area under the ROC curve was 0 .991 ,based the best cut-off point with 4 .5 ,the corresponding diagnostic sensitivity and specificity were 91 .1% and 100% ,respectively . Conclusions Ultrasound image features combined with cesarean section history scores have a high practical value in the diagnosis of placental accreta .

Objective:To explore the application of microvascular flow imaging (MVFI) combined with high-frequency ultrasonography in children with haemophilic arthropathy A (HAA).Methods:Retrospective study.A total of 82 children diagnosed with HAA in the First Affiliated Hospital of Zhengzhou University from October 2018 to October 2020 were recruited.The elbow, knee and ankle joints of each child were examined by high-frequency ultrasonography.The numbers of thickened synovial joints were recorded.Blood flow signals of the thickening of synovial joints was checked by the MVFI and power Doppler ultrasound (PDUS), respectively.Color flow signals were graded by the semi-quantitative scoring systems.The chi- square test and independent multi-group ordinal multi-category rank-sum test were used to compare the differences of MVFI and PDUS in the display of thickened synovial blood flow. Results:A total of 254 joints were involved in 82 children with HAA, including synovial hypertrophy in 188 joints, hydrops articuli in 146 joints, fibrotic septa in 66 joints, cartilage damage in 63 joints, haemosider indeposition in 45 joints, bone erosion in 25 joints, osteophytes in 15 joints and bone remodeling in 8 joints.Grade Ⅱ synovial thickened joints were the most common.The proportion of blood flow signals detected by PDUS in thickened synovial membranes was significantly higher than that detected MVFI (52.66% vs.70.21%, χ2=12.225, P<0.05). Numbers of grade 0 and Ⅰ joints with thickened synovial membranes detected by MVFI were less than those of PDUS, while the opposite result was obtained in detecting grade Ⅱ and Ⅲ joints ( H=21.158, P<0.05). Compared with PDUS, MVFI more sensitively visualized the blood flow of the thickened synovial membrane. Conclusions:MVFI can more prominently detect the thickened synovial blood flow in children with HAA.A combined application of MVFI and high-frequency ultrasonography contributes to the evaluation of children with HAA.

【Objective】 To explore the effect of B lymphocytes on cardiac structure and function and myocardial immune cells during heart development. 【Methods】 Echocardiography, immunofluorescence staining and flow cytometry were used to evaluate the composition of immune cells of the heart and the cardiac structure and function in wild-type (WT) mice and B-lymphocyte-deficient (μMT) mice, respectively. 【Results】 Compared with those of μMT mice, the ratio of heart weight to mouse weight (P<0.05), left ventricular mass (P<0.05) and the cross-sectional area of myocardial cells WT mice were significantly increased, while the ventricular ejection fraction was significantly decreased (P<0.05). The results of mRNA sequencing showed that WT mice and μMT mice differentially expressed genes were mainly enriched in the signal pathway of heart development and hypertrophic cardiomyopathy. The results of flow cytometry showed that WT mice had more Ly6g⁺ neutrophils, CD4⁺ positive T cells (P<0.001) and CD8⁺T cells (P<0.05) compared with μMT mice. 【Conclusion】 B-lymphocyte depletion alters the composition of cardiomyocyte immune cells, reduces left ventricular mass, and increases myocardial contractility.