The observation was carried out on the effects of "Sheng Xue Wan" (pills for promoting the production of blood ) upon the rat colony forming unit--spleen, (CFU-S), colony forming unit--diffusion chamber (CFU-D) and colony forming unit--that responds to erythropoietin(CFB-E)with the extrinsic culture of splenetic colony, the intrinsic culture in medium of a diffusion chamber and the intrinsic plasma clotting. The results of our experiments indicated that "Sheng Xue Wan" has a stimulating action on the multiplication of the rat CFU-S, CFU-D and CFU-E. Thus, we infer that the mechanism of the drug used to treat aplastic anemia might be related to the effect of "Sheng Xue Wan" upon the hematopoietic stem cell.

We investigated the inhibitory effect of autologous sera on erythroid colony forimation(CFU-E) of bone marrow cells from patients with chronic renal failure and the clinical effect of recombinant erythropoietin, Colonies formed in cultures using autologous serum(AS) decreased in 15 cases as compared with those using fetal calf serum(FCS). This inhibitory effect of autologous sera was diminished by treatment with activated charcol in all these cases. The degree of hemoglobin increase after administration of recombinant erythropoietin appeared to correlate with the intensity of inhibitory activity of AS, These data indicate the clinical significance of an inhibitor(S) of erythropoiesis in uremic sera and suggest that the clinical effects of erythropoietin in this disease are further improved if the inhibitor(s) can be effectively removed.

Objective To investigate the pathological characters of capillaries in tubulointerstitial nephropathy,and to study the expression of vascular endothelial growth factor(VEGF)and proliferation cell nuclear antigen(PCNA)in capillary endothelial cells in aristolochic acid-induced nephropathy in rat.Methods 46 male Wistar rats were randomly divided into 2 groups,the model group was composed of 26 rats which were gavaged with the extract of Caulis Aristolochiae Manshuriensis(aristolochic acid AA 20mg/kg?d),and the control group consisted of 20 rats which were treated with equal volume of potable water.The rats were sacrificed in batches at the end of 4th,8th and 12th weeks,and the blood samples were collected from abdominal aorta for the tests of renal functions.The kidney of each rat was harvested.The renal tissues were stained with HE,PAS,and Masson's technique for the analysis of degree of tubulointerstitial injury,interstitial fibrosis,and peritubular capillary(PTC),and the expressions of both VEGF and PCNA were morphologically observed and immunohistochemically analyzed.Results In the model group,the level of serum creatinine/body weight increased markedly,the renal pathological changes consisted of severe acute renal tubulointerstitial damage with cloudy swelling of tubule,degeneration,and exfoliation.With prolongation of feeding time,the damage progressively aggravated,and showed a remarkable tubulointerstitial injury.The interstitial fibrosis area was 31.36% at the end of 12th week.The renal capillaries showed thickening of vessel wall,narrowing of the vessel cavity,obstruction or hyalinization of some capillaries.There was focal infiltration of inflammatory cells around the injured vessels.But there was no apparent change in the globules compared with that in control group.The PTC dwindled in caliber or distorted,and the PTC density decreased significantly in models,especially in the region of tubular damage or interstitial fibrosis.The expression of VEGF showed compensatory up-regulation at the 4th week,but it was down-regulated gradually.The expression of PCNA was up-regulated at the 4th week,but down-regulated after 8th week,and only a few basement membrane naked tubular cells showed positive expression at 12th week.Conclusion AA could induce injury and loss of capillaries of the kidney.The decrease in capillary density might contribute to the impairment of renal function and progressive interstitial fibrosis,and the relative deficiency of VEGF expression may be related to PTC injury,which is one of the causes of chronic progression of aristolochic acid nephropathy.

Objective To explore the effect of VEGF-C,VEGFR-3 gene in primary culture in vitro of laryngeal squamous carcinoma tissue by dexamethasone (DEX). Methods Applying the technique of Drimary tissue cuiture in vitro,twelve specimens,which were identified pathologically as laryngeal squamous cell carcinoma,were cultured.After succeeded culture of twelve specimens,the cells were divided in to two groups.One continued to cuhum,another gave 10-7 mol/L of DEX.VEGF-C and VEGFR-3 gene express in 12 cases of cell were examined by reverse transcription polymerase chain reactions(RT-PCR).Results The frequency of VEGF-C(83%) and VEGFR-3(58%) in primary culture in wtro of laryngeal squamous carcinoma tissue were higher than that of the primary culture in vitro of laryngeal squamous carcinoma tissue by dexamethasone (DEX) (25%,8%P＜0.05).Conclusion The expressions of VEGF-C and VEGFR-3 in Drimary culture in vitro of laryngeal squamous carcinoma tissue by DEX are decreased.It may provide guide to apply DEX in clinic laryngeal squamous carcinoma chemotherapy and laryngeal squamous carcinoma laryngeal obstruction.

Objective To investigate the expression and significance of VEGF and NF-κB/p65 in the two subtypes of pleomorphic adenoma. Methods Immunohistochemistry was applied to detect the expression of VEGF and NF-κB/p65 in pleomorphic adenoma of 60 patients including 31 cases of cell-rich and 29 cases of stroma-rich as well as normal salivary gland tissues of 30 cases from adjacent tumor. Results VEGF positive staining was mainly found in tumor epithelia,while NF-κB/p65 positive was detected in gland alveolus cell and ductal epithelia. The Mean optical density( MA ) values of VEGF were 955.67 ± 305.79,149. 13 ± 60. 85 and 53.46 ± 9. 66, respectively, in cell-rich adenoma, stroma-rich adenoma and normal control. The difference in VEGF expression between the groups was significant (Ps ＜ 0. 05 ) . The MA values of NF-κB/p65 were 529. 80 ± 164. 81,43.40 ±5.46 and 6. 84 ± 1.91 ,respectively,in three groups mentioned above. The difference in NF-κB/p65 expression between the groups was significant ( Ps ＜ 0. 05 ). In pleomorphic adenoma, the expression level of NF-κB/p65 was positively correlated with VEGF. Conclusion ( 1 ) The angiogenesis and proliferation potential of carcinomas increased with the cell component in pleomorphic adenoma. Stroma-poor adenoma is more frequently subjected to malignant transformation than stroma-rich adenoma. (2) NF-κB/p65 may have effects on angiogenesis by activating VEGF. Detecting the expression of VEGF and NF-κB/p65 may be helpful to predict the biological behavior of pleomorphic adenoma and prognosis of the patients, which couldprovide useful information for future targeted therapy of pleomorphic adenoma.

Objective To compare the influence of propofol combined with sufentanil or remifentanil on the quality of wake-up during scoliosis surgery by wake-up test.Methods Fifty pa-tients undergoing scoliosis surgery were randomized into two groups.During the surgery,propofol combined with sufentanily 0.3-0.6 μg·kg-1·h-1 (group SF)or remifentanil 0.2-0.3 μg·kg-1·min-1 (group RF)were continuously infused to maintain anesthesia,and BIS was maintained at 40-60.In wake-up test,the infusion of sufentanyl in group SF was paused and,the infusion rate of remifentanil in group RF was adjusted to 0.05 μg·kg-1·min-1 until the patient completed the wake-up test under instruction.The time that spontaneous breathing occurred,body movement was detected and the capa-bility to follow instructions in both two groups were recorded.MAP,HR,PET CO2 were measured at the time 10 min after medication adjustment (T1 ),waking up(T2 )and 10 min after waking up (T3 ), respectively,in both two groups.Wake-up quality was also recorded.Results The time that sponta-neous breathing occurred,body movement was detected and the capability to follow instructions in group RF were significantly shorter than those in group SF (P <0.05).At T2 the incidence of agita-tion in group RF was significantly higher than that in group SF(P <0.05).And the hemodynamics of group SF were more stable than those of group RF (P <0.05).Conclusion Propofol combined with sufentanil can improve wake-up quality during scoliosis surgery,but the wake-up time is relatively lon-ger.

Objective To investigate the role of human adipose-derived mesenchymal stem cells (ADSC) cultured with astragaloside Ⅳ(Ast) on renal tubular epithelial cells line (HKC) induced by cisplatin in vitro.Methods HKC was divided into four groups:(1) normal control group; (2)cisplatin group; (3) ADSC and HKC group; (4) ADSC cultured with Ast and HKC group.After coincubated with ADSC,the apoptosis index and proliferation of HKC were detected respectively.The levels of interleukin-6 (IL-6),regulated upon activation normal T cell expressed and secreted (RANTES),erythropoietin (EPO) and insulin like growth factor 1 (IGF-1) were detected by ELISA.Transwell culture system were used to test the migration effect of ADSC.Results Compared with cisplatin group,the apoptotic rates of HKC were decreased and the cell number increased obviously (P ＜ 0.05) in ADSC and HKC group and ADSC cultured with Ast and HKC group.Compared with cisplatin group,the expression of IL-6 and RANTES were lower (P ＜ 0.05),and the level of EPO and IGF-1 increased significantly (P ＜ 0.05),in ADSC cultured with Ast and HKC group.Crystal violet staining showed that ADSC cultured with Ast and HKC group migration index was higher than the other three groups (P ＜ 0.05).Conclusion The intervention of Ast may stimulate paracrine effect and migration of ADSC,so as to reduce apoptosis of HKC.

BACKGROUND:The validity of stem celltransplantation for acute kidney injury has been verified by many studies. However, the mechanism of repair of tubular epithelial cellinjury remains unclear. OBJECTIVE:To investigate protective effects and mechanisms of adipose-derived stem cells cultured with astragaloside on the apoptosis of renal tubular epithelial cells induced by cisplatin. METHODS:There were four groups. Renal tubular epithelial cells were induced by 2.5μmol/L cisplatin for 24 hours. Models of renal tubular cellinjury were established in the cisplatin group. Adipose-derived stem cells were cocultured with renal tubular epithelial cells of injured kidney in the adipose-derived stem cells+renal tubular epithelial cells group. Using Transwel chamber, adipose-derived stem cells were incubated with 20 mg/L astragaloside for 48 hours, and then cocultured with renal tubular epithelial cells in the astragaloside-incubated adipose-derived stem cells+renal tubular epithelial cells group. Normal renal tubular epithelial cells served as controls (normal control group). RESULTS AND CONCLUSION:Compared with renal tubular epithelial cellinjury group, AV/PI and TUNEL results demonstrated that the apoptotic rate and number of renal tubular epithelial cells were apparently reduced in the adipose-derived stem cells+renal tubular epithelial cells group and 20 mg/L astragaloside-incubated adipose-derived stem cells+renal tubular epithelial cells group. ELISA results displayed that insulin-like growth factor-1 levels were significantly elevated in the 20 mg/L astragaloside-incubated adipose-derived stem cells+renal tubular epithelial cells group (P<0.05). Western blot assay results exhibited that caspase-3 protein levels were noticeably diminished, but Bcl-2 expression was significantly increased in the 20 mg/L astragaloside-incubated adipose-derived stem cells+renal tubular epithelial cells group (P<0.05). Results suggested that astragaloside-incubated human adipose-derived stem cells suppressed the apoptosis of renal tubular epithelial cells induced by cisplatin, which contributes to the early recovery of injured renal tubule. Its protective mechanism was probably associated with the secretion of insulin-like growth factor-1, inhibition of caspase-3 expression and upregulation of Bcl-2 levels.

Objective To clarity the role of VEGF and its receptors in laryngeal squamous cell carcinoma development and metastasis. Methods The protein and mRNA expression of VEGF and its receptors (Fit-1 and KDR) were examined by Western-blotting and semiquantitative reverse transcriptase polymerase chain reaction analysis in laryngeal squamouse cell carcinoma (20 specimens) and the adjacent non-neoplastic laryngeal tissues (18 specimens). Results The protein and mRNA expression levels of VEGF and its receptors(Fit-1 and KDR) were more abundant in laryngeal squamous cell carcinoma than that in the adjacent non-neoplastic laryngeal tissues (ordedy 4.32±2.21, 2.00±0.91. 1.20±0.55, 0.29~0.31. 2.50±1.69, 0.85±0.28. P<0.01). The expression of VEGF and its receptors(Fit-1 and KDR) in laryngeal squamous cell carcinoma was significantly higher in lymph node positive group than that in lymph node negative group (P<0.01 or P<0.05). Positive correlation was observed between VEGF and KDR in the laryngeal squamouse cell carcinoma and adjacent non-neoplastic laryngeal tissues (P<0.01). There was no correlation between VEGF and Fit-1 expression(P>0.05). Conclusion The intense expression of VEGF, Fit-1 and KDR in laryngeal squamous cell carcinoma provides strong evidence linking VEGF, Fit-1 and KDR expression to the angiogenesis associated with laryngeal squamous cell carcinoma. The positive correlation between VEGF expression and KDR expression clearly demonstrates that KDR is the main signaling receptor for VEGF in laryngeal squamouse cell carcinoma. The VEGF/KDR system plays a functional role in the progression of laryngeal squamous cell carcinoma.

ObjectiveTo study the effect of berberine on diabetes or insulin resistance accompanied with reduction of skeletal moscle and wasting in db/db mice.Methods db/db mouse-a model of diabetes/insulin-resistance was studied,with the wild type mouse as control.After being treated with berberine ( 5 mg · kg-1 · d-1 ) for 3 weeks,the muscle size of tibia anterior (TA) of the animals was measured after staining with laminine/Hyosin Heavy chain-Slow using immunochemistry,then observed under fluorescent microscope and calculated with software.The rate of [ 14 C ] -Phenalanine incorporation into the uuscle was measured to analyze the protein synthesis,and the [ 3 H ] -Tyrosine released into the medium was determined in order to analyze protein degradation.The mRNA expressions of muscle atrophy Fbox-1 ( Atrogin-1 ) and muscle ring finger-1 ( MuRF-1 ) were measured by Northern blot.Results With berberine treatment,blood glucose and fat levels were lowered [ ( 18.55 ± 3.79 vs 26.32 ± 4.02 ) mmol/L,P＜0.01 ; ( 2.75 ± 0.30 vs 3.77 ± 0.52 ) g,P＜0.05 ],but tibia anterior muscle weight/length ratio and cross-section area were decreased,rates of protein synthesis in isolated muscles of db/db mice were decreased by 18％ -22％,and the rates of degradation were significantly raised by 24％ -26％ after berberine treatment.There also was increased the transcription and translation of Atrogin-1 and MuRF-1,accompanied with decreased eukaryotic initiation factor 3 subunit (eIF3-f)protein level simultaneously. ConclusionBerberine improves hyperglycemia and insulin resistance by down-regulating blood sugar and body fat,but it causes reduced protein synthesis and minimally enhanced protein degradation.The mechanism might be related to berberine-induced up-regulating Atrogin-1 、MurF-1 and down-regulating eIF3-f.