Objective To investigate the antitumor effect of cynanauriculoside A(CA) isolated from the root of Cynanchum auriculatum and its effect of apoptosis induction in tumor cells.Methods CA was evaluated for its cytotoxicity in vitro against MCF-7,BEL-7402,and HO-8910 cells by determining MTT assay and its antitumor effects in vivo on S180 tumor-bearing mice by calculating tumor-inhibited rate.Measures of apoptosis including Wright′s-Giemsa staining and flow cytometry(FCM) assay were involved to explore the mechanism.And the toxcity of CA on normal cells was also evaluated on in vitro cultured rat cortical neurons.Results CA showed a definite cytotoxicity to three tumor cell lines with IC50 in the range of 35.68—39.78 mg/L.And it significantly inhibited the tumor growth of S180 tumor-bearing mice at the dose of 40,50,and 160 mg/kg by ig administrated,the inhibitory rates were 20.0%,28.0%,and 48.1%,respectively.At the concentration of 80 mg/L,CA induced obvious apoptosis in MCF-7 cells(P

OBJECTIVE:To establish a method for the contents determination of harpagoside and stilbene glycoside in Shuang-shen xiaolong granule. METHODS:HPLC of harpagoside was performed on the column of Kromasil 100-5 C18 with mobile phase of acetonitrile-1% acetic acid solution (gradient elution) at flow rate of 1.0 ml/min,detection wavelength was 278 nm,column temperature was 25 ℃ and volume injection was 20 μl. HPLC of stilbene glycoside was performed on the column of Kromasil 100-5 C18 with mobile phase of acetonitrile-water(19∶81,V/V)at flow rate of 1.0 ml/min,etection wavelength was 320 nm,column temperature was 25 ℃ and volume injection was 10 μl. RESULTS:The linear range was 0.555 8-8.893 4 μg for harpagoside(r=0.999 9)and 0.010 6-0.340 2 mg for stilbene glycoside(r=0.999 6);RSDs of precision,stability and reproducibility tests were no more than 1.80%;recoveries were 97.30%-101.35%(RSD=1.43%,n=6) and 96.67%-100.83%(RSD=1.48%,n=6),respec-tively. CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the contents determination of harpa-goside and stilbene glycoside in Shuangshen xiaolong granule.

Objective To investigate the effects of pituitary adenylate cyclase activating polypeptide (PACAP) and its receptor (PACAP-R) on the pathogenesis of psoriasis. Methods The expression of PACAP and PACAP-R in the skin from 10 normal controls, 25 psoriatic lesions and non-lesional skins was measured by immunohistochemical technique. Results The expression of PACAP and PACAP-R was significantly lower in the psoriatic lesional skins than that of the non-lesional skins. The area density and mean absorbance of PACAP and PACAP-R in the lesional skins were significantly lower compared with those in the non-lesional skins (P

ObjectiveTo investigate the continuous pathological features of biopsy specimens from five cases of small bowel allotransplantation (SBT) in order to provide more reliable information for the diagnosis and treatment of acute rejection (AR) in SBT.Methods324 biopsy specimens of intestinal mucosa after SBT from 5 patients were collected and studied by histology,histochemistry and electron microscopy.ResultsIn the early stage after operation (0～3 months),AR IND-1 grade was diagnosed for four times on 3 of 5 patients.During 3-6 months,AR IND-1 grade for three times was diagnosed in 2 cases,and AR 2 grade for two times during 7 ～ 12 months. All the patients suffered ischemia reperfusion injury, lymphatic vessel reconstruction and AR.Conclusion The pathological examination of biopsy specimens of intestinal mucosa is still the most reliable detecting method to diagnose AR,and continuous observation may play an important role to monitor the occurrence,development,and treatment response of AR. The final diagnosis of AR depends on structure of intestinal mucosa,crypt epithelium injury and inflammatory cells infiltration. The communication among the pathologist and surgeon is the best way to reduce misdiagnoses.Ultrastructural examination is used to verify the pathogenic microorganism.

ObjectiveTo explore the effects of heat treatment and ultraviolet B (UVB) radiation alone or in combination on the expression of heat shock protein (HSP) 72 in human epidermal melanocytes.Methods Melanocytes were obtained from human foreskin,and subjected to primary culture.After 3 to 5 passages,the melanocytes were classified into 4 groups:control group (receiving no treatment),heat treatment group (treated with heat at 42 ℃ for 1 hour every day for 3 days),UVB group(irradiated with UVB at 50 mJ/cm2 daily for 3days),combination group(treated with heat at 42 ℃ for 1 hour followed by irradiation with UVB at 50 mJ/cm2daily for 3 days).After another 2- to 6-hour culture following the last treatment,melanocytes were collected and subjected to real time PCR and Western blot for the detection of HSP72 mRNA and protein expression,respectively.ResultsThe mRNA and protein expressions of HSP72 were significantly higher in the heat treatment group and combination group than in the control group (mRNA:6.584 ± 0.871 and 7.269 ± 0.454 vs.0.975 ± 0.089,both P ＜ 0.001; protein:2.022 ± 0.058 and 2.080 ± 0.045 vs.0.532 ± 0.033,both P ＜ 0.001 ),but was similar between the UVB group and control group (mRNA:0.832 ± 0.084 vs.0.975 ± 0.089,P ＞ 0.05;protein:0.546±0.021 vs.0.532 ± 0.033,P ＞ 0.05).The ANOVA of factorial design showed that neither heat treatment nor UVB irradiation had interaction effect on the mRNA or protein expression of HSP72 (F =2.106,1.399 respectively,both P ＜ 0.05).ConclusionsHeat treatment can cause an increase in the expression of HSP72,which may enhance the function of melanocytes and protect melanocytes from UVB induced damage.

ObjectiveTo investigate the effect of heat treatment combined with narrow band ultraviolet B(NB-UVB) on cultured normal human melanocytes in vitro.MethodsMelanocytes were isolated from the foreskin of normal human,cullured in vitro,and irradiated with NB-UVB of different doses(20,30,50,70,90,120 and 180 mJ/cm2).Then,MTT assay was performed to evaluate the proliferation and activity of melanocytes to determine the optimal dose of UVB for the next experiment.Melanocytes were classified into 3 groups to be treated with heat at 42 ℃ for 1 hour (heat group),irradiated with UVB at 50 mJ/cm2 (UVB group),or irradiated with UVB at 50 mJ/cm2 followed by heat treatment at 42 ℃ for 1 hour (combination group),daily for 3 successive days; those receiving no treatment served as the control.After 24-hour culture following the last treatment,tyrosinase activity was evaluated with L-dopa as the substrate,melanin content was detected by NaOH assay,and cell cycle stages were determined by flow cytometry.ResultsNB-UVB irradiation decreased the viability of melanocytes in a dose-dependent manner,and the optimum dose of UVB was 50 mJ/cm2.The tyrosinase activity of melanocytes was 0.244 ± 0.018 and 0.310 ± 0.015 respectively in the UVB group and combination group,and increased by 3.8％ (P ＜ 0.05) and 31.9％ (P ＜ 0.05) respectively compared with the control group (0.235 ± 0.018); the melanin content was 0.201 ± 0.016 and 0.286 ± 0.019,respectively in the UVB group and combination group,and increased by 17.5％ (P ＜ 0.05 ) and 67.3％ (P ＜ 0.05) compared with the control group (0.171 ± 0.016).In comparison with the control group,the percentage of melanocytes in G1 phase was decreased by 23.94％ in the UVB group(P＜ 0.05) and 33.51％ in the combination group(P ＜ 0.05),while that in S phase and G2 phase increased by 15.35％ (P ＜ 0.05 ) and 11.93％ (P ＜ 0.05),respectively in the UVB group,and 17.76％ (P ＞ 0.05) and 16.08％ (P ＞ 0.05),respectively in the heat group.ConclusionHeat treatment and NB-UVB can synergistically enhance the tyrosinase activity and accelerate melanogenesis,proliferation and differentiation,of melanocytes.

ObjectiveTo observe the increasing effect of infrared irradiation on tyrosinase activity and melanin content in cultured normal human epidermal melanocytes in vitro and to explore the optimal dose of infrared irradiation.MethodsEpidermal melanocytes were isolated from normal human foreskin tissue,and subjected to primary culture.Methyl thiazolyl tetrazolium(MTT) assay was performed to evaluate the effect of different doses(0,20,60,80,100,140,240,320 J/cm2)of infrared light on the proliferation of melanocytes and to select the optimal irradiation dose.Then,melanocytes were irradiated with infrared light at the optimal dose for 3 consecutive days followed by the determination of tyrosinase activity,melanin content,and cell cycle via dopa oxidation assay,NaOH solubilization method and flow cytometry,respectively.ResultsThe best intervention dose of infrared light was 80 J/cm2.The tyrosinase activity(A492 nm) and melanin content(A492 nm)were 0.3601 ± 0.0301 and 0.2748 ± 0.0243 respectively in melanocytes after irradiation with infrared light of 80 J/cm2 for 3 days,significantly higher than those in unirradiated melanocytes(0.3114 ± 0.0341,0.2325 ±0.0254,respectively,both P ＜ 0.05),with an increase rate of 15.64％ and 18.19％ respectively.Cell cycle analysis revealed a decline in cell percentage in G1 phase(P ＜ 0.01 ) but a concomitant increase in cell percentage in G2 and S phase (both P ＜ 0.05) in irradiated melanocytes compared with unirradiated melanocytes.ConclusionsThe optimal dose of infrared light is 80 J/cm2 for the irradiation of melanocytes,and this dose of infrared light can increase melanin content,tyrosinase activity,differentiation and proliferation of melanocytes.

Objective To investigate the clinical presentation, endoscopy and pathological features of subclinical cellular rejection (SCR) of small bowel allotransplantation. Methods Three times of SCR in a patient after isolated small bowel transplantation were studied by endoscopy and microscopy, and the clinical data and literature were reviewed. Results SCR was an unusual type of acute rejection after small bowel transplantation. SCR showed low-grade morphological changes of acute rejection, and may be relived after low-dose steroid or bolus steroid was given. Conclusion The causes of SCR are not clear now. SCR may be the early stage of clinical acute rejections, and may be correlated with unexpected high grade acute rejection, and chronic loss function of graft. The biopsy through ileoscopy is a "golden standard" of diagnosis of SCR in small bowel transplantation.However, the vessel lesions of graft, ileus, and inflammation should be excluded before diagnosis.

Objective There are a few reports on rats with spontaneous congenital cataract in China .The purpose of this study was to investigate the morphological changes of lens in rats with spontaneous congenital cataract . Methods 24 d, 1-year rats with cataract and microphthalmos cataract and normal rats (n=5) were selected as research objects .Their lens were observed by a slit lamp microscope and taken photos in front of them , followed by examination through light micrograph and transmission electron micros-copy. Results Rats with microphthalmos cataract showed narrowed palpebral fissure and broaden nucleus while rats with cataract showed normal palpebral fissure and narrowed nucleus .As for 24 d,1-year rats with microphthalmos cataract , the fibers of their lens showed derangement and vacuole-like degeneration by light microscope , in addition, the abnormal connection between fiber cells were observed by electron microscopy .As for 1-year normal rats , the fibers were in consistent structure and regular arrangement without cell ingredient . Conclusion The appearance and morphological changes of the lens in rats with spontaneous congenital cataracts are in consistence with the pathological changes of cataracts , which is appli-cable in further research on the pathogenesis of cataract .

Purpose To investigate the expression and significance of HLA-G in ovarian serous carcinoma ( OSC) . Methods HLA-G antigen was immunohistochemically labeled on paraffin-embedded sections of 108 OSCs. The relationship between HLA-G expression and the clinicopathologic parameters was studied. Results The positive expression of HLA-G was observed in 58. 33% (63/108) of OSC tissues. Positive expression of HLA-G was significantly related with lymph node metastasis, recurrence, occurrence site, FIGO stage and MDACC grading system (P<0. 05). In survival analysis, the expression of HLA-G was significantly relevant to prognosis (P=0. 015). Multivariate Cox analysis showed the expression of HLA-G was an important prognosis factor of OSC (P=0. 01). Con-clusion The positive expression of HLA-G could predict high grade and advanced stage of OSC as well as poor prognosis. Also it could distinguish high-grade OSC from low-grade OSC.