OBJECTIVETo study the effect of Yiqi Wenyang Huoxue Lishui principle (YWHL, a therapeutic principle with Chinese medicine for reinforcing Qi warming Yang, activating blood circulation and promoting urination) on some neuroendocrine factors in patients with congestive heart failure (CHF).

METHODSForty-nine patients of CHF with heart function of grade III-IV were randomly divided into 2 groups. The 29 cases in the treated group were treated with YWHL and the 20 cases in the control group treated with captopril. Changes of angiotensin II (AT II), atrial natriuretic peptide (ANP), endothelin (ET), nitric oxide (NO) and alpha-granule membrance protein -140 (GMP-140) were observed.

RESULTSAfter treatment, plasma level of AT II, ANP, ET, and NO lowered in both groups with insignificant difference, but the GMP-140 reduced more significantly in the treated group (P < 0.05).

CONCLUSIONYWHL showed a regulatory effect of neuroendicrine system partially similar to that of angiotensin-converting enzyme inhibitor, it possibly can improve the ventricular remodeling and would be beneficial to prevent the thrombus formation and improve heart failure by means of inhibiting platelet activity.

Adult ; Aged ; Angiotensin II ; blood ; Atrial Natriuretic Factor ; blood ; Coronary Disease ; complications ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Heart Failure ; drug therapy ; etiology ; physiopathology ; Humans ; Male ; Middle Aged ; Neurosecretory Systems ; drug effects ; P-Selectin ; blood ; Phytotherapy

This study investigated a nano drug delivery system built by one sort of modified trimethyl chitosan (TMC). The TMC was modified by cRGDyk, ligand of integrin receptor avβ3. Single factor screening was used to optimize the prescription in which the particle sizes of TMC nanoparticle (TMC NPs) and cRGDyk modified TMC nanoparticle (C-TMC NPs) were (240.3 ± 4.2) nm and (259.5 ± 3.3) nm. Electric potential of those two nanoparticles were (33.5 ± 0.8) mV and (25.7 ± 1.6) mV. Encapsulation efficiencies were (76.0 ± 2.2) % and (74.4 ± 2.0) %. Drug loading efficacies were (50.1 ± 2.1) % and (26.1 ± 1.0) %. Then the cellular uptake, uptake mechanism and transport efficacy of TMC NPs and C-TMC NPs were investigated using Caco-2 cell line. The uptake rate and accumulating drug transit dose of C-TMC NPs were 1.98 and 2.84 times higher than TMC NPs, separately. Mechanism investigations revealed that caveolae-mediated endocytosis, clathrin-mediated endocytosis and macropinocytosis were involved in the intercellular uptake of both TMC NPs and C-TMC NPs. What is more, free cRGDyk could remarkably inhibit the uptake of C-TMC NPs.
Biological Transport ; Caco-2 Cells ; Caveolae ; Chitosan ; chemistry ; Clathrin ; Endocytosis ; Humans ; Integrin alphaVbeta3 ; chemistry ; Nanoparticles ; Particle Size ; Pinocytosis

Objective To explore the influencing factors and possible mechanism of osteonecrosis of the femoral head(ONFH)in patients with leukemia after allogeneic hematopoietic stem cell transplantation (Allo-HSCT).Methods One hundred and two patients with leukemia who received Allo-HSCT between January 2003 and October 2007 were evaluated for ONFH and graft-versus-host disease(GvHD)within 2years after transplantation,and the dosage of methylprednisolone(MP)in every patient from the first diagnosis of leukemia to 2 years after Allo-HSCT was calculated.For patients who suffered acute GvHD(aGvHD) and chronic GvHD(cGvHD),the serum leveh of soluble ligand of receptor activator of nuclear factor kappa B (sRANKL)which was index for differentiation of osteoclast(OC),tartrate-resistant acid phosphatase-5b(TRAP-5b)and C-terminal telopeptide of collagen Ⅰ(CTP-Ⅰ)which both were indexes for metabolism of OC were detected by enzyme-linked immunosorbent assay in different stages of MP dosage.According to these results.possible factors for ONFH after Allo-HSCT were analyzed.Results Seven in 102 patients after Allo-HSCT had experienced ONFH within 2 years,the incidence was 6.9%(7/102),which was not related to age,gender,types of leukemia and donor.ONFH mainly developed in patients with aGvHD and cGvHD,and the incidence could be achieved to 21.9%(7/32)which Was much higher than that in patients with no GvHD,merely aGvHD or merely cGvHD(P<0.05).In patients with aGvHD and cGvHD,the average dosage of MP in ONFH(+)was(232.7±28.6)mg/kg which was extremely higher than that in ONFH(-)(115.1±16.9)mg/kg(P<0.05).The serum levels of sRANKL,TRAP-5b and CTP-Ⅰ were also higher when ONFH happened than ahead of pretreatment and 28 days after transplantation(-)(P<0.05),and they were closely related to the dosage of MP(correlation coefficient was 0.597,0.664 and 0.682 respectively,P<0.05).Conclusions After Allo-HSCT,ONFH is related to the dosage of MP in treatment of GvHD.MP may be responsible for enhancement of OC differentiation and metabolism in leukemia patients,and ultimately induced the onset of ONFH.

Objective To compare the clinical efficacy and complications of allogeneic peripheral blood stern cell transplantation (Allo-PBSCT) and immunosuppressive therapy (IST) for severe aplastic anemia(SAA). Methods Twenty-five patients with SAA underwent allogeneic HLA-matched sibling donor PBSCT(n = 12) and IST (n = 13). PBSCT group received conditioning regimen of cyclophosphamido(Cy) in combination with antithymocyte globulin(ATG). IST group received ATG followed by cyclosporine A (CsA).The short-term and long-term effects and complications were investigated. Results The mean time of recovery in the absolute neutrophil count (ANC), platelet and hemoglobin (Hb) in PBSCT group [(13.5±2.3), (23.5±4.1), (82.7±6.1)d, respectively]was shorter than those in IST group [(32.6±3.5), (73.8±6.2), (296.4±12.5)d, respectively]and there were statistical differences between two groups(P<0.05). But the one-year treatment effect between two groups showed no difference (P>0.05). There were no statistical differences in 3-year survival and overall survival rate between two groups (P>0.05). However, statistical difference was observed in overall relapse rate (P<0.05). The common complication in two groups was virus infection including cytomegalovirus (CMV) and varicella zoster virus (VZV), but there was no statistical difference in the incidence of virus infection between them (P>0.05). Conclusions Both allo-PBSCT and IST are effective methods for treating patients with SAA. PBSCT is considered preferentially in clinic, because of its advantages in faster hematopoietic engraftment, lower relapse rate and no increased complication.

Objective To study the NR5A1 gene mutation in patients with idiopathic hypogonadotropic hypogonadism(IHH), and to find the new mutation point.Methods Sixty-one IHH patients and 100 normal control subjects were collected and genomic DNA was extracted from blood samples.These patients were with normal karyotype and no abnormality was discovered in magnetic resonance imaging (MRI) scan of the pituitary.Other endocrine diseases were also excluded.The 2-7 exons and splice-sites of NR5A1 gene were amplified with polymerase chain reaction.DNA of the coding sequence and splice-sites of NR5A 1 were sequenced by double deoxidizing terminal end sequencing method in 61 IHH and 100 normal control subjects.The results of sequencing were compared with their corresponding sequence data.61 IHH kindreds were investigated and the clinical data of these patients were collected.Finally, the phenotype and genotype positive cases were analyzed.Results Six patients carried NR5A1 gene mutational sites in 61 cases of IHH.Analysis of sequencing results from 100 age and ethnicity matched control subjects did not show any of these novel changes.Conclusions One mutation in NR5A1 gene may affect protein structure and function, which should be considered in male IHH patients with normal karyotype and without insufficiency of adrenal function.

This study investigated a nano drug delivery system built by one sort of modified trimethyl chitosan (TMC). The TMC was modified by cRGDyk, ligand of integrin receptor avβ3. Single factor screening was used to optimize the prescription in which the particle sizes of TMC nanoparticle (TMC NPs) and cRGDyk modified TMC nanoparticle (C-TMC NPs) were (240.3 ± 4.2) nm and (259.5 ± 3.3) nm. Electric potential of those two nanoparticles were (33.5 ± 0.8) mV and (25.7 ± 1.6) mV. Encapsulation efficiencies were (76.0 ± 2.2) % and (74.4 ± 2.0) %. Drug loading efficacies were (50.1 ± 2.1) % and (26.1 ± 1.0) %. Then the cellular uptake, uptake mechanism and transport efficacy of TMC NPs and C-TMC NPs were investigated using Caco-2 cell line. The uptake rate and accumulating drug transit dose of C-TMC NPs were 1.98 and 2.84 times higher than TMC NPs, separately. Mechanism investigations revealed that caveolae-mediated endocytosis, clathrin-mediated endocytosis and macropinocytosis were involved in the intercellular uptake of both TMC NPs and C-TMC NPs. What is more, free cRGDyk could remarkably inhibit the uptake of C-TMC NPs.

Objective To investigate the effect of bortezomib on the proliferation and apoptosis in leukemia cell line NB_4-R2 in vitro and provide some new evidences for the treatment of acute promyelocytic leukemia APL with ATRA-resistant using bortezomib. Methods NB_4-R2 cells were incubated with bortezomib at different does for 48 h. The proliferation capacity was measured by MTT assay, the morphology of cell apoptosis observed with Hoechst33342 staining by fluorescence microscopy and the percentage of apoptosis calculated by flow cytometry. The expression of apoptosis protein of cleaved (poly ADP-ribose polymerase, PARP) and Caspase-3 were determined by Western blotting. Results The proliferation of NB_4-R2 cells were obviously inhibited by bortezomib in vivo and the role of inhibition was a does-dependant manner within the scope of the bortezomib concentration from 1-5 μg /L.The incidence of inhibition was up to 74.9 % at the bortezomib concentration of 5 μg/L. Within this scope of the bortezomib concentration mentioned above, the role of inhibition of proliferation of NB_4-R2 cells mainly showed an increase of the late apoptosis, and the percentage of apoptosis was up to 78.9 %. In the meaning time, the expressions of the apoptotic protein of cleaved PARP and Caspase-3 were up-regulated in NB_4-R2 cells after treated with bortezomib by Western blotting assay. Conclusion Bortezomib can inhibit the proliferation of NB_4-R2 cells in vivo by inducing cell apoptosis.

AIM: To re-identify the special motif regulating osteoclast(OC)differentiation in receptor activator of nuclear factor kappa B(RANK)to provide evidences for studying the mechanism of OC differentiation.METHODS: Eight amino acids were mutated(from DIIVVYVS into ELLAAFAA)in the fragment between the 533th and the 540th amino acids in RANK cytoplasmic domain.Eight mutant TNFR1/RANK chimeras,each consists of TNFR1(tumor necrosis factor receptor 1)extracellular domain linked to transmembrane domain and cytoplasmic domain of RANK with one amino acid mutated in cytoplasmic domain was constructed by point mutation method.After the eight mutant chimeras were finished,they were packed with plat E cell line to produce the retrovirus expressing mutant TNFR1/RANK.The bone marrow macrophages(BMMs),isolated from TNFR1/R2 double knockout mice,were infected with retrovirus derived from different mutants and infected BMMs which did not differentiated into OCs were inspected after stimulated by TNF-? and M-CSF.The fragment consisted of different amino acids in TNFR1/RANK chimeras,which couldn't induce OC formation after mutated,may be the special motif regulating OC differentiation.RESULTS: We found that all BMMs transfected by TNFR1/RANK-533,TNFR1/RANK-539 or TNFR1/RANK-540 differentiated into OCs,indicating that none of amino acids D533,V539 or S540 had an effect on OC differentiation.A fewer of BMMs transfected by TNFR1/RANK-534 differentiated into OCs,indicating that I534 had a partial effect on OC formation.Most importantly,BMMs transfected TNFR1/RANK-535,TNFR1/RANK-536,TNFR1/RANK-537 or TNFR1/RANK-538 did not differentiated into OCs,indicating each of amino acids I535,V536,V537 and Y538 played a pivotal role in OC differentiation.CONCLUSION: The amino acid fragment consists of I534,I535,V536,V537 and Y538 may be the special motif regulating OC differentiation in RANK.

Objective To know the prevalence of iodine deficiency disorders(IDD) and the implementation of control measures against the disorders in high-risk areas of Hainan province. Methods Typical sampling principle was used. Eight townships with lower coverage rate of iodized salt were chosen in IDD high-risk counties of Hainan. New cretinism cases were screened among children aged 10 years and under. Intelligence quotient (IQ)value, urinary iodine (UI) concentration and thyroid gland size (by palpation and B ultrasonic) were investigated in children aged 8 to 10 years and questionnaire was also carried out among students from grade 5 in elementary school. Edible salt and urine of women of childbearing age were collected to detect iodine levels, respectively,questionnaire was also carried out among the women of childbearing age. The UI was determined by As3-Ce4+ catalytic spectrophotometer, the salt iodine level was determined using self-quantitative kit. IQ values were measured by the Combined Raven Test in China (CRT-C2). Results Seventy-two townships were investigated in three consecutive years, 7937 children aged 8 to 10 years, 1797 women of childbearing age and 4128 students of grade 5 were included in the survey. One child was diagnosed as suspected new cretinism case. The coverage rate of iodized salt was increased from 44.6%(629/1411) in 2007 to 92.1%(1688/1832) in 2009. The goiter rate was dropped from 5.9% (269/4548), 6.0% (274/4548) in 2007 to 1.6% (24/1461), 0.1% (2/1461) in 2009 bypalpation, and by B-ultrasound, respectively, in children aged 8 to 10 years. The median urinary iodine(MUI) was found to be increased from 97.4 μg/L in 2007 to 165.0 μg/L in 2009, and the percentage of UI less than 50.0 μg/L, and 100.0 μg/L were dropped from 21.4%(973/4548), 51.2%(2329/4548) in 2007 to 7.5 %(110/1461), 23.4%(342/1461) in 2009, respectively. The MUI of women of childbearing age was found to be elevated from 73.7 μg/L in 2007 to 126.1 μg/L in 2009, and the value was also increased from 55.7 μg/L to 121.5 μg/L in pregnant women, but these values were still lower than 150.0 μg/L The pass rate of the questionnaire of IDD controlling among the students of grade 5 and the women of childbearing age was raised from 29.8% (446/1495) ,24.1% (179/742) in 2007 to 65.8%(732/1112), 72.1%(264/366) in 2009, respectively. The mean IQ of children was 90.4±16.0, and the percentage of IQ value less than 69 was 12.6% (1000/7937). Conclusions The diseased state of IDD has improved in the high-risk counties of disorders in Hainan province, but the pregnant women are still iodine deficient. Iodine supplimentation should be promoted among pregnant women.

Objective To investigate the value of ultrasound in tethered cord syndrome (TCS) in infants. Methods From December 2005 to July 2013, 25 cases TCS were confirmed by surgery in Shenzhen children's hospital. The ultrasonogram and MRI of 25 infants were analysed retrospectively. The diagnostic accuracy of ultra-sound and its clinical significance were evaluated. Results In 25 cases of TCS confirmed by surgery, the coincidence rate of conus level detected by ultrasound [96%(23/24)] was lower than that by MRI (100%(25/25)). Ultrasound showed reduced spinal cord vibration in 4 cases, disappeared spinal cord vibration in 21 cases. Two cases of reduced spinal cord vibration detected by ultrasound were confirmed as disappeared spinal cord vibration by surgery. The coincidence rate of disappeared spinal cord vibration detected by ultrasound was 91%(21/23). The sacral spinal seg-ments form showed by ultrasound were entirely consistent with those of MRI, including 4 cases of enlarged spinal cord, 13 cases of spinal cord without enlargement and 8 cases of spinal cord ended with rat caudate. Twenty-five cases of TCS had malformations:7 cases meningocele (3 cases combined lipoma), 14 cases myelomeningocele (5 cases combined lipoma, 1 case combined hydromyelia), 3 cases spinal canal-epidermis fistula (all combined lipoma) and 1 case solitary lipoma. Compared with the operation findings, ultrasound misdiagnosed 2 cases of myelomeningo-cele as meningocele, missed 1 of case lipoma which combined with meningocele. MRI missed two cases of spinal canal-epidermis fistula. Conclusions Infantile spinal ultrasound examination can accurately locate the position of conus, accurately display the spinal cord. Compared with MRI, ultrasound examination can real-time visually display spinal cord vibration and help to diagnose tethered cord. Ultrasound examination are convenient, repeatable opera-tion with low cost, therefore it can be used as the preferred screening method to diagnose of tethered spinal cord.