ObjectiveTo explore the procedures and methods for genetic diagnosis in one nonsyndromic variants of congenital neutropenia (NSVCN) patient and its pathogenic mutation.Methods Genomic DNA was prepared from one NSVCN patient who had progressed to chronic myelomonocytic leukemia and ELA2,HAX1,WASp and GFI1 genes were amplified and sequenced.Results A novel compound heterogeneous mutation consisting of two frame-shift mutations (c.430-1insG and c.655- 9del5bp) was found in HAX1 gene.ConclusionA practically genetic diagnosis procedure for NSVCN has been established,and the novel HAX1 gene mutation may contribute to the etiology of NSVCN.

Objective To investigate the effects of Ulinastatin(UTI)on the function of splenic lymphocytes from rats with severe acute pancreatitis(SAP).Method Twenty-eight Wister rats(clean grade)were randomly divided into control,sham operation,SAP,and ulinastatin group.No operation was performed in control group.And rats with sham-operation received laparotomy and catheterization into choledocho-pancreatic duct without injection of sodium deoxycholic.Rats in ulinastatin group received ulinastatin injection(50000 U/kg)via tail vein 30 minutes after pancreatitis induced with DCA injected into pancreatic duct.Rats ofother groups were given equal volume of saline.At 2,4 hours after operation,all animals were killed by neck dislocation,and splenocytes were isolated and cultured in RPMI 1640 medium containing 10%fetal calf serum.Proliferation of splenecytes was determined with MIT cellular proliferation assay.Levels of Th1 cytokines(IL-2,IFN-γ)and Th2 cytokine(IL-10)in supematants of splenoeytesweremeasured by ELISA.Quantitative data were expressed as mean±SE.Statistical analyses were performed by Student's t test with SPSS software(version 10.0 for Windows).A P value less than 0.05 Was considered statistically significant. Results The concentration of IL-2, IL-10 and IFN-γ and proliferative activity of splenocytes in SAP group were significantly lower than that in sham operation group.In contrast,the proliferative as well as the eytokine-releasing capacities of the solenecms from rats treated with UTI were significantly increased compared with those from rats with SAP.Conclusions The deficiencies in proliferation and cytokine release in response to antigen stimulation inaplys an anergic state of splenocytes during SAP.Treatment with UTI contributed to the recovery of the immune function by improving proliferative responses and cytokine release of splenocytes.

Aim To investigate the effect of polydatin on cardiomyocyte hypertrophy induced by high glucose (25.5 mmol·L -1 )and insulin (0.1 μmol ·L -1 ) (HGI)and its possible influence on peroxisome prolif-erator-activated receptor-β (PPARβ)/nuclear tran-scription factor-κB (NF-κB)/nitric oxide (NO)signa-ling pathway.Methods The cardiomyocyte hypertro-phy was characterized in rat primary cardiomyocytes by measuring the cell surface area,protein content,and atrial natriuretic factor (ANF)mRNA expression.The mRNA and protein expressions were measured by qRT-PCR and Western blotting,respectively.The activity of NO synthase (NOS)and NO content were measured by reagent kit through ultraviolet spectroscopy.Results HGI significantly induced cardiomyocyte hypertrophy which increased the cell surface area,protein content and ANF mRNA expression (P <0.01 ).Meanwhile, the expressions of PPARβmRNA and protein reduced while the NF-κB p65 and iNOS expressions increased significantly which occurred in parallel with rising NOS activity and NO concentration (P <0.01 ).Polydatin (0.1,1,10 μmol·L -1 )inhibited the cardiomyocyte hypertrophy induced by HGI (P <0.01 ),and re-versed the mRNA and protein expressions of PPARβ, NF-κB p65 and iNOS,and NOS activity,as well as NO content.These effects of polydatin were abolished by GSK0660 (1 μmol·L -1 ),a selective PPARβan-tagonist (P <0.05 ).Conclusion Polydatin resists HGI-induced cardiomyocyte hypertrophy,which may be mediated by PPARβup-regulation,and then NF-κB-iNOS-NO pathway inactivation.

AIM:To investigate the role of peroxisome proliferator-activated receptor β( PPARβ)-nitric oxide (NO) signal pathway in cardiomyocyte hypertrophy induced by high glucose (25.5 mmol/L) and insulin (0.1 μmol/L) ( HGI) .METHODS: The cardiomyocyte hypertrophy was characterized in rat primary cardiomyocytes by measuring the cell surface area, protein content, and the mRNA expression of atrial natriuretic factor (ANF).The mRNA and protein ex-pression were measured by real-time PCR and Western blotting , respectively .The activity of NO synthase ( NOS) and NO content were measured by a reagent kit through ultraviolet spectroscopy .RESULTS:HGI induced profound change of hy-pertrophic morphology , and significantly increased the cell surface area , protein content and mRNA expression of ANF (P<0.01), but decreased the expression of PPARβat mRNA and protein levels (P<0.05).At the same time, the ex-pression of inducible NOS (iNOS) was obviously elevated (P<0.01), which occurred in parallel with the rising NOS ac-tivity and NO concentration (P<0.01).GW0742 (1 μmol/L), a selective PPARβagonist, inhibited the cardiomyocyte hypertrophy induced by HGI ( P<0.01 ) , and up-regulated the expression of PPARβat both mRNA and protein levels . Meanwhile, GW0742 also inhibited the increases in iNOS expression , NOS activity, and NO content induced by HGI , which were abolished by GSK0660 (1 μmol/L), a selective PPARβantagonist (P<0.01).CONCLUSION: PPARβdown-regulation and the following iNOS-NO activation are involved in the cardiomyocyte hypertrophy induced by HGI .

The aim of this paper is to compare the cytotoxicity and cellular uptake efficiency of three kinds of poly(b-benzyl-L-amino) block-poly(ethylene glycol) nanoparticles (PXA-PEG-NPs) using Calu-3 cells, and select one as a nasal drug delivery vector for curcumin (Cur). Poly(gamma-benzyl-L-glutamate) block-poly(ethylene glycol) nanoparticles (PBLG-PEG-NPs), poly(gamma-benzyl-L-lysine) block-poly(ethyleneglycol) nanoparticles (PZLL-PEG-NPs) and poly(gamma-benzyl-L-aspartate) block-poly(ethylene glycol) nanoparticles (PBLA-PEG-NPs) were prepared by emulsion-solvent evaporation method. MTT assays were used to evaluate the cytotoxicity of PXA-PEG-NPs against Calu-3 cells. The cellular uptake of nanoparticles was visualized by an inverted fluorescence microscope and quantified by a flow cytometer. The results indicated that even at high concentration of 2 mg x mL(-1) the three nanoparticles had no cytotoxicity on Calu-3 cells. Compared to the curcumin solution, the three curcumin-loaded PXA-PEG-NPs showed significantly higher cellular uptake efficiency on Calu-3 cells (at equal concentration of curcumin with 5 microg x mL(-1) Cur solution), PBLG-PEG-NPs group was the highest. The cellular uptake increased with incubation time, and has positive correlation with nanoparticle concentration. In brief, PXA-PEG-NPs are conducive to delivery Cur into cells, and PBLG-PEG-NPs might be provided as a good nasal drug delivery carrier.

Objective: To observe the clinical efficacy of DZWJY-1 type electronic moxibustion apparatus and traditional moxibustion in treating knee osteoarthritis (KOA). Methods: A total of 76 eligible patients were randomized into an electronic moxibustion apparatus group and a traditional moxibustion group, with 38 cases in each group. The electronic moxibustion apparatus group was intervened by DZWJY-1 type electronic moxibustion apparatus, and the traditional moxibustion group received moxa stick moxibustion for treatment. Neixiyan (EX-LE 4), Dubi (ST 35), Xuehai (SP 10) and Liangqiu (ST 34) were selected for both groups and the treatment was conducted 3 times a week for a total of 12 times. The visual analog scale (VAS) and the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) scores were observed before treatment and after 6 and 12 sessions of treatment, respectively. Results: There were 4 dropout cases in the traditional moxibustion group. Therefore, this trial had 72 valid cases, including 38 cases in the electronic moxibustion apparatus group and 34 cases in the traditional moxibustion group, the differences in the baseline data between the two groups were statistically insignificant (P>0.05). After 6 and 12 sessions of treatment, the VAS scores decreased significantly with the increase of treatment sessions in both groups (all P<0.01), and the betweengroup differences were statistically insignificant at the same time points (both P>0.05). The pain intensity was evaluated using the weighted value of VAS score. The markedly effective rate was 47.4％ and the total effective rate was 89.5％ in the electronic moxibustion apparatus group, versus 50.0％ and 94.1％ in the traditional moxibustion group, and the betweengroup differences were statistically insignificant (both P>0.05). After 6 and 12 sessions of treatment, the total score and the component scores including pain, stiffness and difficulty moving in the WOMAC decreased significantly with the increase of treatment sessions in both groups (all P<0.01), and the between-group differences were statistically insignificant (all P>0.05). Conclusion: Electronic moxibustion apparatus and traditional moxibustion both are effective in reducing joint pain and improving joint function in KOA patients, and they are equivalent comparing the clinical efficacy.

OBJECTIVETo explore the in vitro anti-tumor effect and mechanism of dendritic cell (DC) tumor vaccine induced by astragalus polysacharin (APS).

METHODSPeripheral blood mononuclear cells (PBMCs) isolated from human peripheral blood. DCs obtained from human peripheral blood were cultivated and added with culture solution for in vitro inducing them to immature DCs. On the 5th day of culture, 100 microg/mL (as the final concentration) APS was added to cells in the APS group. DCs were induced to mature in the cytokine groups by adding 20 ng/mL rhTNF-alpha (as the final concentration). Changes of morphology and phenotype of DCs were observed. Mature DCs were sensitized with tumor antigen SGC-7901 and co-cultured with allogeneic T cells. The proliferative function of T lymphocytes was detected by MTT assay. Levels of IL-12 and IFN-gamma in co-cultured supernatant were detected by ELISA. Cytotoxic lymphocytes (CTL) activated by DC were co-cultured with tumor cell SGC-7901. The specific killing capacity of CTL to target cells was detected by LDH release assay.

RESULTSThe morphological observation and phenotypic identification of APS induced DCs were in accordance with the characteristics of mature DCs. APS induced mature DCs could stimulate the proliferation of allogeneic T lymphocytes. The proliferation index of T cells increased with increased ratio of stimulator cells to effector cells (P < 0.05). Levels of IL-12 and IFN-gamma in co-culture supernatant significantly increased in a time-dependent manner (P < 0.05). CTL cells activated by sensitization of DCs could significantly kill tumor cells, and the killing effect increased along with increased effector-to-target ratio.

CONCLUSIONAPS could in vitro induce DCs to mature, promote its antigen-presenting capacity, effectively activate CTLs, and enhance anti-tumor function of the organism.

Antigen-Presenting Cells ; cytology ; drug effects ; immunology ; Cancer Vaccines ; immunology ; Cell Line ; Cell Proliferation ; drug effects ; Coculture Techniques ; Dendritic Cells ; cytology ; drug effects ; immunology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Interferon-gamma ; immunology ; Interleukin-12 ; immunology ; Leukocytes, Mononuclear ; cytology ; immunology ; Lymphocyte Activation ; T-Lymphocytes, Cytotoxic ; cytology ; drug effects

Objective To explore the clinical skills and curative efficacy of Glubran (NBCA-MS) embolization of cerebral arteriovenous malformations. Methods Ninety-six cases of cerebral arteriovenous malformations were treated by 187 endovascular embolizations with different concentrations of Italy GEM company's Glubran (NBCA-MS). Follow-up was performed to 96 cases of cerebral arteriovenous malformation with NBCA-MS embolization. Results During the follow-up in 96 cases, clinical symptoms completely disappeared in 92, were improved in 3 and reoccurred in 1. The malformation was totally got rid of in 44 of 48 cases DSA examinated, and the 80%-90% embolization was achieved in the rest who need further γ-knife therapy. In all cases, γ-knife therapy was performed in 32 cases after embolization, and among them 26 cases received DSA examination 1-2 years after γ-knife therapy, and 21 cases were proved to have no malformation. Conclusions The method ofendovascular emdolization with NBCA-MS is safe, reliable and effective to treat the cerebral arteriovenous malformation.

Objective To explore brain activity features during the resting state in alcohol dependent individuals,and study the relationship between the brain activity features and alcohol dependent individuals' clinical symptoms.Methods Twenty-four alcohol dependent individuals and 22 healthy control subjects,well matched in gender,age,education and handedness,were enrolled as the alcohol dependent group and control group respectively.AGE 3.0 T MR scanner was used to acquire all the subjects' resting state data. DPARSF software was used to process resting functional MRI data,and then the whole brain fractional amplitudes of low frequency fluctuation (fALFF) data were acquired.Two-sample t test statistical analysis was made to access fALFF difference between the two groups. Results In comparison with the control group,the alcohol dependent group showed reduced fALFF in bilateral medial prefrontal gyrus,right inferior occipital gyrus,left precuneus,left inferior temporal gyrus,and left posterior lobe of cerebellum (0.64-1.69 vs.0.87-1.78,t =- 4.23- - 2.79,P ＜ 0.05 ). fALFF was increased in the alcohol dependent group at the anterior cingulate,bilateral inferior frontal gyrus,right middle frontal gyrus,bilateral insular lobe,bilateral dorsal thalamus ( 0.86-1.82 vs. 0.76-1.58,t =3.56-3.96,P ＜ 0.05 ).Conclusion Alcohol dependent individuals had abnormal activity at the bilateral prefrontal lobe,anterior cingulate,bilateral dorsal thalamus,bilateral insular lobe,left posterior lobe of cerebellum et al,during the resting state,and these abnormal activities might be related with clinical manifestation and pathophysiology.

OBJECTIVETo explore the effect of Qiling Decoction (QD) combined highly active antiretroviral treatment (HAART) on expression levels of peripheral blood Th17 and Treg cells in HIV/AIDS patients.

METHODSTotally 55 HIV/AIDS patients were randomly assigned to the treatment group (28 cases) and the combination group (27 cases). Besides, 21 HIV negative patients were recruited as the healthy control group. Those in the treatment group received HARRT alone, while those in the combination group received HAART combined QD. The observation lasted for 24 weeks. Meanwhile, according to peripheral blood CD4+ T cell counts before treatment, HIV/AIDS patients were assigned to three subgroups. For patients in subgroup 1, 1 cells/microL < CD4+ T cell counts < or = 100 cells/microL; For patients in subgroup 2, 101 cells/microL < CD4+ T cell counts < or = 200 cells/lL; For patients in subgroup 3, 201 cells/microL < CD4+ T cell counts < or = 350 cells/microL. Expression of peripheral blood Th17 and Treg cells, and number of CD4+ T cell counts were detected using flow cytometry (FCM)in HIV/AIDS patients at the pre-treatment baseline, week 4, 12, and 24, as well as those in the healthy control group.

RESULTSCompared with the healthy control group, CD4+ T cell counts and the baseline expression level of Th17 cells in the peripheral blood of HIV/AIDS patients significantly decreased, the expression level of Treg cells significantly increased P < 0.01). Compared with before treatment in the same group, CD4+ T cell counts all increased at week 4, 12, and 24 in the two treatment groups, showing statistical difference (P < 0.05, P < 0.01). There was no statistical difference in the effective rate at various CD4+ T cell levels between the two groups (P > 0.05). There was no statistical difference in expression levels of Th17 and Treg cells between the combination group and the treatment group at any time point (all P >0.05). The Th17/Treg ration significantly increased in the combination group after 24 weeks of treatment, showing statistical difference when compared with the treatment group (U = 2.135, P = 0.038).

CONCLUSIONQD could improve the immune balance of Th17/Treg cells, which might be one of its mechanisms for improving HIV/AIDS patients' immunity.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adult ; Antiretroviral Therapy, Highly Active ; CD4 Lymphocyte Count ; Case-Control Studies ; Drugs, Chinese Herbal ; therapeutic use ; Female ; HIV Infections ; drug therapy ; immunology ; Humans ; Male ; Middle Aged ; Phytotherapy ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology