BACKGROUND:Under ischemia/hypoxia microenvironment,very low survival rate of transplanted bone marrow mesenchymal stem cells (BMSCs) in the host limits its efficacy in the treatment of acute myocardial infarction.OBJECTIVE:To explore the tolerance of human heme oxygenase 1 (hHO-1) gene modified rat BMSCs to ischemia/hypoxia injury.METHODS:The rat BMSCs were transfected with hHO-1 recombinant adenovirus.Western blot assay was used to determinate the optimal time of hHO-1 protein expression.hHO-1 modified rat BMSCs were cultured in hypoxia and serum-free conditions that simulated ischemia/hypoxia microenvironment in vivo.Cell counting kit-8 and trypan blue staining were performed to detect the survival rate of BMSCs at 12,24,48,72 hours after culture under the ischemia/hypoxia microenvironment.Flow cytometry was used to detect apoptosis in BMSCs at 24 hours after culture under the ischemia/hypoxia microenvironment.RESULTS AND CONCLUSION:The expression of hHO-1 protein was highest at 4 days after transfection.Under the ischemia/hypoxia microenvironment for 12,24,48,72 hours,the survival rates of transfected BMSCs were significantly higher as compared with the untransfected cells (P ＜ 0.05),shown by the cell counting kit-8 and trypan blue staining.In addition,the results from flow cytometry showed that there was a higher survival rate of transfected BMSCs than untransfected cells at 24 hours of culture under the ischemia/hypoxia microenvironment (P ＜ 0.05).To conclude,hHO-1 modified rat BMSCs have stronger tolerance to the ischemia/hvpoxia microenvironment.

BACKGROUND AND PURPOSE: Five single-nucleotide polymorphisms (SNPs) (rs4379368, rs10504861, rs10915437, rs12134493 and rs13208321) were recently identified in a Western population with migraine. These migraine-associated SNPs have not been evaluated in a Han Chinese population. This study investigated the associations of specific SNPs with migraine in a Han population. METHODS: This was a case-control study of Han Chinese residing in Fujian Province. Polymerase chain reaction—restriction-fragment-length polymorphism analysis and direct sequencing were used to characterize the relationships of SNPs in a control group of 200 subjects and in a migraine group of 201 patients. RESULTS: The frequencies of the five SNPs did not differ between patients with migraine and healthy non migraine controls. However, subgroup analysis indicated certain SNPs were more strongly associated with migraine with aura or migraine without aura than with controls. The CT genotype of rs4379368 was more common in migraine patients with aura (75%) than in migraine patients without aura (47.9%) and controls (48.5%) (p<0.05), and the TT genotype of rs10504861 was more common in migraine patients with aura than in controls (8.3% vs. 0.5%) (p<0.05). Meanwhile, the CC genotype of rs12134493 was less common in migraine patients without aura than in controls (80.6% vs. 88%) (p<0.05). CONCLUSIONS: Our findings suggest that the rs4379368 and rs10504861 SNPs are markers for susceptibility to migraine with aura and that rs12134493 is a marker for the risk of migraine without aura in this Han population. Future studies should further explore if these associations vary by ethnicity.
Asian Continental Ancestry Group* ; Case-Control Studies ; Epilepsy ; Genotype ; Humans ; Migraine Disorders* ; Migraine with Aura ; Migraine without Aura ; Polymorphism, Single Nucleotide

Objective To explore the expression level of CD36 in peripheral blood mononuclear cells(PBMCs) from patients with crytococcal meningitis and its clinical significance.Methods The experimental group was the peripheral blood samples from 36 patients diagnosed as crytococcal meningitis in Changhai Hospital and Changzheng Hospital in Shanghai from September 2013 to February 2017,while the control group was the peripheral blood samples from 36 health individuals examined at the same time.PBMCs were separated by density gradient centrifugation method,the relative mRNA expression of CD36,TLR4 in PBMCs was tested by qRT-PCR,the expression level of cytokines such as TNF-α in plasma was tested by ELISA,the comparison of the examined values of experimental and control groups performed by two independent samples' t test,Pearson correlation analysis was performed on the relative mRNA expression of CD36 in PBMCs and cytokines such as TNF-α in experimental group.Results The relative mRNA expression of CD36 in PBMCs of experimental group was 2.01 ± 0.63,the control was 1.49 ± 0.47,and there was statistical difference (t =3.969,P =0.000 2).Pearson analysis results showed that in experimental group CD36 was positively related with the protein expression level of IFN-γ in plasma (r=0.384,P＜0.05) and negatively related with TGF-β(r=-0.487,P＜0.005).Conclusion CD36 might involve in the immunopathology process of crytococcal meningitic by influencing the expression of cytokines such as IFN-γ,TGF-β,and was the potential target for monitoring and treating this disease.

Objective To study the expression of CD69 in exosomes existed in serum from patients with autoimmune pancrea titis (AIP) and primarily discuss its affection on the pathology of disease progression.Methods Serum samples from 35 cases diagnosed as AIP and 35 healthy individuals examined at the same time were collected during October 2012 to December 2016 in Changhai Hospital,they were defined as experimental and control groups,separately.The cytokine levels in serum were measured by ELISA.The expression of surfaced molecules in exosomes existed in serum was tested by magnetic bead conjunct antibody method in flow cytometry.The comparison between the two groups' measurement data was measured by two independent samples' t test,correlation between two variables was showed by Pearson coefficient.Results The flow cy tometry analysis showed that the expression level of CD69 in serum exosomes in experiment and control groups were (85.76 ±19.45 vs 17.01±5.89)％,with statistical difference(t=20.01,P＜0.0001).CD69+ exosomes in experiment group were positively related with serum IL-4,IFN-γ and TGF-β (r 0.456,0.678,0.548,all P＜0.05),while CD69 exosomes were negatively related with serum IL-4,IFN-γ,IL-10 and TGF-β (r=0.589,-0.399,-0.784,-0.657,all P＜0.05).Conclusion The expression of CD69 in exosomes might participate in AIP progression through influencing the function of CD4 +T cell,and it was the potential examined biomarker and therapeutic target.

Objective To prepare F7 thermosensitive liposome and evaluate its physicochemical properties, then investigate its cytotoxicity against tumor cells in vitro. Methods The F7 thermosensitive liposome was prepared by the pH gradient active drug loading method using dipalmitoyl phosphatidylcholine myristoyl lyso-phosphocholine and 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy (polyethylene glycol)-2000 as membrane materials. The encapsulation efficiency and drug loading were determined for the F7 thermosensitive liposome by HPLC. The phase transition temperature of F7 thermosensitive liposome was investigated by differential scanning calorimetry;the liposome morphology was observed by atomic force microscopy;the drug release of liposome was examined by dialysis;and the particle size and zeta potential were measured through Malvern particle size analyzer. The cytotoxicity of F7 and F7 thermosensitive liposome was determined by the MTT method, and the freeze-drying process was optimized using the designexpert software. Results The encapsulation efficiency of F7 thermosensitive liposomes was (97.56±0.22) ％, and the drug loading ratio was (1.51±0.01) ％. The phase transition temperature of F7 thermosensitive liposome was 39.9℃, the zeta potential was (-15.10±0.85) mV, the particle size was (86.94±1.21) nm, and the poly disperse coefficient was 0.17±0.01. Compared with the F7 injection, the F7 thermosensitive liposomes showed a stronger, dose-dependent inhibitory effect on the growth of lung cancer H1299 and breast cancer MCF-7 cells. The freeze-dried powder of liposomes dissolved well with the encapsulation efficiency of 95％ and the particle size of approximately 130 nm. Conclusion The F7 thermosensitive liposome prepared by the pH gradient active drug loading method has high encapsulation efficiency and good stability. The preparation method is simple and feasible for further development of the F7 preparation.

Objective To examine the expression of CD93 in serous exosomes from patients with cryptococcal meningitis,fur-ther explore its clinical significance.Methods The 38 experimental serum samples were from patients who received the diag-nosis with cryptococcal meningitis in Changhai Hospital and Changzheng Hospital in Shanghai from November 2012 to De-cember 2016.The diagnosis standardization was that the cerebrospinal fluid dyeing was positive or the culturing was posi-tive.The 38 controls were collected from the health individuals examined at the same time.The exosomes in serum was sepa-rated by the ultracentrifuge method.Magnetic bead-capture combined with the flow cytometry method was used to identify antigens on the surface of exosomes.The protein level of cytokines in serum was quantified by ELISA method.The compari-son of tested values from experimental and controlled groups was measured by two independent samples't test,and the cor-relation between two variates was showed by Pearson coefficient.Results The results of flow cytometry showed that the ex-pression of CD93 on exosomes in the serum samples from experimental group was higher than that in the controlled group, which was(79.11±19.31 vs 23.98±6.56)%,with difference in statistics(t=16.66,P<0.000 1).The expression levels of IFN-γin serum samplefrom experimental and control groups were(39.78±10.77 vs 58.98±16.99)pg/ml,with differ-ence in statistics(t=5.884,P<0.000 1).The expression levels of IL-17 in serum sample from experimental and control groups were(16.32±4.03 vs 3.11±0.87)pg/ml,with difference in statistics(t=19.75,P<0.000 1).The expression lev-els of IL-1β in serum sample from experimental and control groups were(57.12 ± 12.98 vs 13.45 ± 4.78)pg/ml,with difference in statistics(t=19.46,P<0.000 1).The percentage of CD93+exosomes was positively correlative with IFN-γ, IL-17,IL-1β(r=0.488,0.456,0.532,P<0.01)in serum samples from the experimental group and there was statistical difference.Conclusion In the disease process of cryptococcal meningitis,the expression of CD93 on exosomes might involve in the diseases'processes by affecting the CD4+T cells.

To study the absorption kinetics of paeoniflorin lipid liquid crystalline nanoparticles (Pae-LLCN) in different intestinal segments of rats and compare them with paeoniflorin(Pae) solution. Rat everted gut sac models were adopted for intestinal absorption test, and Pae content was determined by HPLC method to study the absorption characteristics of Pae-LLCN in rat duodenum, jejunum, ileum and colon, and investigate the effects of different drug concentrations on intestinal absorption. Results showed that Pae-LLCN and Pae were well absorbed at different intestine segments and different concentrations. The absorption constant Ka was increased with the increasing of the drug concentration, indicating possible passive absorption. The accumulative absorption volume Q and absorption constant Ka of Pae-LLCN were higher than those of Pae at each intestinal segment(P<0.05). The results revealed that Pae-LLCN and Pae could be well absorbed in whole intestinal segments and its mechanism may be passive absorption. LLCN can effectively improve the intestinal absorption of Pae.

OBJECTIVE: To analyze the hematological characteristics of Hb Broomhill and Hb Hornchurch, and prenatal diagnosis should be carried out in two families. METHODS: RBC parameters and hemoglobin electrophoretogram were analyzed on the peripheral blood of all patients, and amniotic fluid was collected for prenatal diagnosis. PCR-Flow fluorescent hybridization and Sanger sequencing were performed for gene diagnosis of thalassemia. RESULTS: Three cases of Hb Broomhill were detected, including 2 cases with common SEA α-thalassemia, which was characterized by hypochromic microcytic mild anemia, the capillary electrophoregram revealed a tiny shoulder peak before the Hb A peak; 1 case was diagnosed as Hb Hornchurch combined with β-thalassemia, which also showed mild anemia. Hemoglobin electrophoretogram showed an abnormal hemoglobin variant peak at Hb A CONCLUSION The carriers of Hb Broomhill and Hb Hornchurch do not have microcytic hypochromic anemia, which do not aggravate the hematological symptoms, such as anemia when being combined with thalassemia of the same type.
Anemia, Hypochromic ; Hemoglobins, Abnormal/genetics* ; Heterozygote ; Humans ; alpha-Thalassemia/genetics* ; beta-Thalassemia

OBJECTIVE: To analyze the hematological characteristics of Chinese METHODS: Hemoglobin electrophoresis and blood routine test were used to analyze the hematological indexes of all peripheral blood samples，PCR-Flow fluorescent hybridization and Gap-PCR were used to detect the globin gene mutations and the data were analyzed statistically. RESULTS: The 3 types of deletion β- Thalassemia patients were showed as hypochromic small cell anemia. The MCH and MCV values of Taiwan type β-thalassemia patients were the lowest. The results of hemoglobin electrophoresis showed that the increasing of HbF was found in all of the 3 types. Except for the decreasing of Hb A2 in Chinese CONCLUSION Through analyze the hematological characteristics, it can be provide that the guidance for the differential diagnosis and genetic consultation of the three commonest deletion β-thalassemia in Chinese.
China ; Diagnosis, Differential ; Fetal Hemoglobin ; Humans ; Mutation ; Thalassemia ; beta-Thalassemia/genetics*