Objective:To explore the influence of tumor cells on CD4~+ CD25~+ Treg cells via TLRs.Methods:The numbers changes of CD4~+ CD25~+ Treg cells in the co-culture system of Lewis lung cancer cells and splenic lymphocytes were detected by flow cytometry;The expression of Foxp3 and TLR1-9 mRNA after co-culture were detected by RT-PCR;TLR9 expression on Lewis lung cancer cells was blocked by TLR9 receptor antagonist chloroquine.Results:Compared with control group,the number of CD4~+ CD25~+ Treg cells and Foxp3 mRNA expression were significantly increased in the co-culture group (P＜0.05).The expression of a variety of TLRs were affected by the co-culture of lymphocytes with Lewis lung cancer cells,and TLR9 mRNA expression was significantly increased compared with that of the control group (P＜0.05);Blocking TLR9 of Lewis lung cancer cells significantly reduce CD4~+ CD25~+ Treg cells and Foxp3 mRNA (P＜0.05).Conclusion:Lewis lung cancer cells could increase both number and function of CD4~+ CD25~+ Treg cells through inducing TLR9 expression in immunocells.This might be one of mechanisms of tumor-induced immune tolerance,and by which to contribute to the occurrence and progression of tumors.

Objective:To explore the influence of tumor cells on CD4+CD25+Treg cells via TLRs.Methods:The numbers changes of CD4+CD25+Treg cells in the co-culture system of Lewis lung cancer cells and splenic lymphocytes were detected by flow cytometry;The expression of Foxp3 and TLR1-9 mRNA after co-culture were detected by RT-PCR;TLR9 expression on Lewis lung cancer cells was blocked by TLR9 receptor antagonist chloroquine.Results:Compared with control group,the number of CD4+CD25+Treg cells and Foxp3 mRNA expression were significantly increased in the co-culture group(P

BACKGROUND: Osteosarcoma is the most common type of bone tumor and generally occurs between the age of 10 to 25; moreover, in clinical practice osteosarcoma is found to occur more often in minority nationalities in Xinjiang Autonomous Region. Do its incidence and prognosis vary between nationalities at gene levelOBJECTIVE: To observe the different expressions of p53, bcl-2 and proliferative cell nucleus antigen (PCNA) gene during the development of osteosarcoma between various Xinjiang nationalities.DESIGN: Non-randomized comparative experiment taking clinical pathological specimens as subjects.SETTING: Immunohistochemical Laboratory, the Pathological Center of Xinjiang Medical University.PARTICIPANTS: Totally 52 pieces of specimen were obtained from excised osteosarcoma tissues in the Pathological Department, the First Affiliated Hospital of Xinjiang Medical University, and the Pathological Department, the First Affiliated Hospital of Shihezi University, between January 1,1984 and December 31, 2001. The 52 cases of osteosarcoma included 29cases derived from male patients and 23 from female patients; among them there were 12 cases of Kazak minority, 17 cases of Uygur minority and 23cases of Han nationality. Other 32 specimens were obtained from 32 patients with tumor-like lesions (such as osteofibrous dysplasia or fibrous dysplasia), including 7 cases of Kazak minority, 11 cases of Uygur minority,and 14 cases of Han nationality. The informed consent was obtained from the patients.METHODS: This experiment was carried out at the Immunohistochemical Laboratory, the Pathological Center of Xinjiang Medical University. LSAB method was used to detect p53, bcl-2 and PCNA expression in the two groups. The first antibody was re placed by PBS as blank control, and the available positive expression was taken as positive control. P53 protein and PCNA were observed to express in cell nucleus, appearing obvious redbrown granules with positive expression, whereas Bcl-2 protein was expressed in cytoplasm.lationship between the expression of Bcl-2, p53 and PCNA in osteosarcoma.of different nationalities: The expression of p53, bcl-2 and PCNA in Kazak minority, Uygur minority and Han nationality was not remarkable p53, bcl-2 and PCNA in osteosarcoma and bone tumor-like lesions: The expression of P53, Bcl-2 protein and PCNA in osteosarcoma was remarkably higher than that in tumor-like lesions (42.31% vs. 3.13 %, 59.62%sion of bcl-2, p53 and PCNA in osteosarcoma: There was a close correlation between bcl-2 and p53, as well as between bcl-2 and PCNA in osteosarcoma tissues (X2 =5.818 2, 4.900 0, P＜0.05).CONCLUSION: The results indicate that there was no statistical difference between various nationalities in the expression of p53, bcl-2 and PCNA, as well as osteosarcoma differentiation. Suggesting that these genes may share the common regulation during the development of osteosarcoma,which is less associated with their nationality-related hereditary background.

Objective To assess the efficacy and safety of two different protocols of ketogenic diet (KD)-eating on demand or eating at regular intervals for refractory epilepsy in children.Methods Sixty children with refractory epilepsy were randomly divided into eating on demand group (n =30) and eating at regular intervals group (n =30) by random number table method.After taking the whole amount of KD,the capillary blood ketone and glucose level and urine ketone were monitored every 6 hours in 72 continuous hours.Seizure frequency and onset time were recorded.Antiepileptic efficacy and diet tolerability of the two groups were evaluated on 4 weeks,12 weeks,24 weeks and 48 weeks after initiating the diet.Adverse effects were monitored.Results After treatment of 4 weeks,the complete seizure remission rates of eating on demand group and eating at regular intervals group were 33.3％ (10/30) and 30.0％ (9/30) respectively,which suggested a comparable efficacy for two groups (P ＞ 0.05).The day when KD started to work was averaged (6.18 ± 2.42) d and (8.63 ± 2.63) d respectively.The group of eating on demand showed a faster onset of action (P ＜0.05).After treatment of 12 weeks,24 weeks and 48 weeks,complete seizure remission rates of eating on demand group were 30.0％ (9/30),34.8％ (8/23) and 36.8％ (7/19) respectively;the eating at regular intervals group were 33.3％ (10/30),30.4％ (7/23) and 44.4％ (8/18) respectively.The two groups had no significant difference (P ＞ 0.05).One year later,the treatment retention rates of the two groups were 63.3 ％ (19/30) and 60.0％ (18/30) respectively.There was no significant difference (P ＞ 0.05).The adverse effects mainly including transient gastrointestinal symptoms and metabolic disturbances were mostly tolerable and curable.Conclusion The two different protocols of KD-eating on demand and eating at regular intervals are both effective and well-tolerated for refractory epilepsy in children.While protocol of eating on demand is more easier to achieve ketotic state and the effect is more quickly,so it can be more easily received by children.Therefore in clinical practice,we can choose flexible eating time according to children's eating habits,which can improve the therapeutic compliance.

Objective To investigate the clinical and genetic characteristics in a patient with 17β-hydroxy-steroid dehydrogenase (17β-HSD) 3 deficiency, regarding its pathophysiology and pathogenesis. Methods Clinical features and laboratory data were analyzed in a pedigree of 17β-HSD3 deficiency. Blood samples from the patient and his parents were collected. HSD17B3 gene was screened for mutations by PCR and subclone sequencing. Results The patient presented with pubertal virilization and gynecomastia. The physical examination showed female external genitalia and testes in inguinal canals. The chromosome karyotype was 46, XY. Serum FSH, LH, dehydroepiandrosterone sulfate, androstenedione and 17-OH-progesterone levels were raised, whereas plasma testosterone was lowered. Sequencing analysis revealed 4 nucleotide deletion (172-175del) of HSD17B3 gene. Conclusion Virilization and gynecomastia in puberty suggest the probability of 17β-HSD deficiency. It may be verified clinically by hCG-stimulating test and confirmed by gene diagnosis.

BACKGROUND: Growth factors and other biological methods have become very popular in the repair of degenerative interevrtebral disc. Insulin-like growth factor-1 (IGF-1) can promote the proliferation of nucleus pulposus cells, and synthesis of functional extracellular matrix, but the mechanisms remain unclear.OBJECTIVE: To investigate the effect of IGF-Ⅰ on the expressions of aggrecan and collagen type Ⅱ in nucleus pulposus cells, and to explore its signal transduction mechanism.METHODS: The human nucleus pulposus cells were isolated and cultured. Passage 3 nucleus pulposus cells were induced in different concentrations of IGF-1 (0, 20, 50, 100 and 200 μg/L), respectively. The expressions of aggrecan and collagen type Ⅱ were detected by reverse transcription PCR and western blot assay. Western blot assay was adopted to observe the effect of 100 μg/L IGF-1 on the activation of PI3K/Akt signaling pathway in nucleus pulposus cells,and the expression of aggrecan and collagen type Ⅱ was detected after the inhibition of PI3K/Akt pathway by LY294002.RESULTS AND CONCLUSION: With the increase of IGF concentration, the expression levels of aggrecan and collagen type Ⅱ were increased gradually. 100 μg/L IGF-1 could significantly promote the expressions of p-PI3K and p-Akt (P <0.01), while LY294002 reversed this effcet (P < 0.01). 100 μg/L IGF-1 significantly upregulated the expression levels of aggrecan and collagen type Ⅱ in nucleus pulposus cells (P < 0.01); in contrast, LY294002 significantly downregulated the expression levels of aggrecan and collagen type Ⅱ promoted by IGF-1(P < 0.01). These results indicate that IGF-1 can promote the expression levels of aggrecan and collagen type Ⅱ in nucleus pulposus cells via the PI3K/Akt signaling pathway.

Objective To investigate the related risk factors of contrast-induced acute kidney injury (CI-AKI)after cerebrovascular intervention. Methods The clinical data of 5423 patients performed cerebrovascular angiography and intervention at the Departments of Neurology and Neurosurgery,Guangdong People′s Hospital from January 2005 to December 2013 were analyzed retrospectively. The patients who underwent cerebrovascular angiography and intervention were evaluated and screened. A clinical history database was established. All the selected patients used iodixanol,an isotonic contrast agent. The occurrence of CI-AKI was used as an endpoint. The patients were divided into either a CI-AKI group or a non CI-AKI group. A multivariate Logistic regression model was used to analyze the risk factors associated with the occurrence of CI-AKI. Results A total of 4164 patients were finally enrolled,including 137 had CI-AKI. The incidence of CI-AKI was 3. 3%. The results of multivariate Logistic regression showed that age >60 years (OR,1. 965,95%CI 1. 244-3. 136),baseline estimated glomerular filtration rate (eGFR)<60mL/(min·1. 73 m2)(OR,4. 163,95%CI 2. 422-5. 873),diabetes (OR,3. 140,95%CI 1. 983-3. 902),and anemia (OR,1. 524,95%CI 1. 226 -3. 253)were the influencing factors for occurring CI-AKI after cerebrovascular angiography and intervention. Conclusion Chronic kidney disease (eGFR<60 mL/[min·1. 73 m2 ]),diabetes,anemia,and old age (age >60 years)are the independent risk factors for occurring CI-AKI after cerebrovascular angiography and intervention.

Objective To compare the clinical and electroencephalogram(EEG)characteristics and therapeutic response prognosis of different age groups of children with epileptic spasm .Methods From January 2002 to October 2011 the clinical data ,EEG fea-tures of epileptic spasms children under 15 years old with unknown disease cause(cryptogenic) were retrospectively reviewed .74 of them were followed up for 12 to 92 months .All of them were divided into two groups on the basis of onset age :3 -12 months of onset as group infantile-onset spasms(group IOS ,n=60);and 12 months to 7 years old of onset as group late-onset epileptic spasms (group LOS ,n=14) .Clinical process ,seizure semiology and EEG features were compared between two groups .Results Semiologic features of two groups were similar ,but they showed differences in interictal EEG features including the background ,the location of discharges ;The response to drugs between the two groups are also different .Conclusion There are differences between group IOS and group LOS when comparing EEG features and response to drugs .

Objective To observe the effect of berberine on glucose transport in 3T3-L1 adipocytes and to investigate its mechanism. Methods The glucose consumption of the cells was determined by the glucose oxidase method. The glucose transportation rate of the cells was assayed by the uptake of 2-deoxy-〔 3H〕-D-glucose. Protein kinase B (Akt) activity was detected by immunoprecipitation and Western blot. The gene expression of c-Cbl-associated protein (CAP) was detected by Northern blot. Results 0.1～200 ?mol/L berberine significantly increased glucose consumption in 3T3-L1 adipocytes with a dose-dependent effect, which was independent of insulin. The glucose transportation was significantly increased in adipocytes incubated with 0.1～10 ?mol/L berberine; the action began at 2 h and reached a peak value at 12 h. The results of immunoprecipitation and Western blot showed that berberine did not enhance Akt activity. The result of Northern blot indicated that berberine significantly decreased CAP mRNA expression. Conclusion Adipocytes are the important target cells of berberine. Berberine significantly increases glucose transportation and consumption in adipocytes, the action appeares to be independent of insulin signal pathway.

OBJECTIVE To generate hemophilia A (HA) patient-specific inducible pluripotent stem cells (iPSCs) and induce endothelial differentiation. METHODS Tubular epithelial cells were isolated and cultured from the urine of HA patients. The iPSCs were generated by forced expression of Yamanaka factors (Oct4, Sox2, c-Myc and Klf4) using retroviruses and characterized by cell morphology, pluripotent marker staining and in vivo differentiation through teratoma formation. Induced endothelial differentiation of the iPSCs was achieved with the OP9 cell co-culture method. RESULTS Patient-specific iPSCs were generated from urine cells of the HA patients, which could be identified by cell morphology, pluripotent stem cell surface marker staining and in vivo differentiation of three germ layers. The teratoma experiment has confirmed that such cells could differentiate into endothelial cells expressing the endothelial-specific markers CD144, CD31 and vWF. CONCLUSION HA patient-specific iPSCs could be generated from urine cells and can differentiate into endothelial cells. This has provided a new HA disease modeling approach and may serve as an applicable autologous cell source for gene correction and cell therapy studies for HA.
Cell Differentiation ; Hemophilia A ; pathology ; therapy ; urine ; Humans ; Induced Pluripotent Stem Cells ; cytology ; transplantation ; Urine ; cytology