OBJECTIVE:To prepare dexibuprofen sustained-release pellets,and to analyze the drug release behavior in vitro. METHODS:Centrifugal granulation powder layering-eudragit dispersion coating method was used to prepare dexibuprofen sus-tained-release pellets using 3%HPMC as adhesive agent. The formula of the pellets was optimized by orthogonal test with weight ra-tio of sucrose to dexibuprofen,weight ratio of HPMC to Eudragit NE30D and coating weight as factors,using 1,4 and 10 h accu-mulated release rate (Q) as index. The release of the drug from the pellets was analyzed. RESULTS:The optimized formulation was that the proportion of sucrose to drug was 1:10,the weight ratio of HPMC to Eudragit NE30D was 1.5:1,the increased weight of coating material was 8%. Q1 h,Q4 h and Q10 h of prepared pellets were 21%,57% and 89%,respectively(n=3). The co-rrelation coefficient of zero-order,one-order and Higuchi equation release model were 0.956 6,0.989 9,0.996 5. CONCLUSIONS:Prepared pellets show good sustained-release effect in vitro. Drug release of pellets is more in accordance with Higuchi equation.

Objective:The effects of IL-13 ( Interleukin-13 ) on SDF-1 ( Stromal cell derived factor 1 ) and EGF ( Epidermal growth factor) expression in fibroblasts co-cultured with breast cancer cells were investigated to explore the mechanism for IL-13 in the development of breast cancer.Methods:The co-culture of human breast cancer cell line MDA-MB-231 with the human skin fibroblast line CCC-ESF-1 ( ESF) was used in vitro and in tumor-burdened nude mice.The effects of IL-13 on SDF-1 and EGF expression in the co-cultured fibroblasts in vitro were analyzed using reverse transcription quantitative polymerase chain reaction ( RT-qPCR ) , flow cytometry and Western blot assay.The proliferation of the co-cultured human breast cancer cells in vitro was detected by Cell counting kit-8(CCK-8).The effects of IL-13 on SDF-1 and EGF expression in the fibroblasts of tumor tissue of tumor-burdened nude mice were analyzed using immunofluorescence and laser confocal microscope, and the tumor volumes were examined.Results: IL-13 could up-regulate SDF-1 and EGF expression in the fibroblasts co-cultured with breast cancer cells in vitro,and promoted the proliferation of the co-cultured breast cancer cells.In tumor-burdened nude mice,IL-13 enhanced SDF-1 and EGF expression of fibroblasts in tumor tissue, and accelerated tumor growth.Conclusion:IL-13 up-regulates SDF-1 and EGF expression of fibroblasts co-cultured with breast cancer cells.The molecular mechanism of promoting effect of IL-13 on breast cancer relates to SDF-1 and EGF of fibroblasts in breast cancer stroma.

Purpose　To observe the changes of dynorphin-like immunoreactivities of neurons in some rat brain nuclei that are related to analgesia following exogenous administration of melatonin.　Methods　The experimental rats were divided into two groups, injected intraperitoneally with melatonin 110 mg/kg and with vehicle, respectively. One hour after the injection, the rat brain was processed for coronal sections. The sections were stained with immunohistochemical ABC technique. The integral optical density (IOD) of the stained section was measured by the computer-assisted image processing technique.　Results　Dynorphin-like immunoreactivities in the supraoptic nucleus and nucleus raphe dorsalis showed obvious reduction following the single injection of melatonin.IOD values in the above nuclei were decreased significantly (P<0.01) with the melatonin treatment. In the paraventricular nucleus of hypothalamus, periaqueductal gray and nucleus raphe magnus, there was no difference (P>0.05) about the IOD values between melatonin-treated group and vehicle-treated group.　Conclusions　Melatonin may result in the decrease of dynorphin content in the supraoptic nucleus and nucleus raphe dorsalis.

Objective To study the influence of residual methylene blue after plasma viral inactivation on the human immune cell function by using the peripheral blood mononuclear cell(PBMC) .Methods PBMC were isolated by adopting the Ficoll‐Hypaque density gradient centrifugation method and co‐cultured for 72 h in presence of specific T cell stimulating factors(Anti‐CD3/28 and Anti‐CD28) ,with or without different concentration of methylene blue .The culture supernatant was collected and detected the cyto‐kines secretion situation by ELISA .After 66 h culture ,CCK‐8 dye was added and continueously cultured for 4-6 h ,the prolifera‐tion was determined at A450 .Results The high‐concentration doses of methylene blue (1 .25 ,2 .5 ,5 μmol/L groups) had signifi‐cantly inhibiting effect on the proliferation of PBMC stimulated by Anti‐CD3/28(P< 0 .01) ,its OD value was decreased from 0 .897 ± 0 .385 to 0 .632 ± 0 .334 ,0 .524 ± 0 .254 and 0 .445 ± 0 .287 respectively ,showing certain dose dependent effect .The high concentrations of methylene blue (1 .25 ,2 .5 ,5 μmol/L groups) could down‐regulate interleukin(IL)‐17a ,IL‐10 and interferon (IFN)‐γ secreted by anti‐CD28 induced PBMC ,moreover showing a dose dependent effect .1 .25 ,2 .5 ,5 μmol/L methylene blue af‐fected the IL‐17a level secreted by PBMC from (406 ± 57)pg/mL descending to (276 ± 38) ,(192 ± 31) ,(134 ± 24)pg/mL respec‐tively ;affected PBMC to secrete IL‐10 ,its level was reduced from (184 ± 15) pg/mL to (132 ± 13) ,(110 ± 12) ,(42 ± 8)pg/mL ;af‐fected PBMC to secrete IFN‐γ,its level was deduced from (4 512 ± 187)pg/mL to (2 876 ± 143) ,(2 234 ± 153) ,(1 988 ± 112)pg/mL respectively .Conclusion High concentrations of methylene blue (≥1 .25 μmol/L ) has the significant inhibiting effect on the proliferation and cytokine secretion functions of PBMC .In other words ,the residual methylene blue concentration in viral inactiva‐tion plasma (≤0 .33 μmol/L) has no obvious effect on the immune function of PBMC ,but whether this concentration of methylene blue having the effect on human pure T cell immune function needs to be further evaluated and studied .

AIM: To investigate the influence of the cornus officinalis glycosides (COG) on immunological function of corneal transplantation model of rats, and to clarify the immunosuppressive mechanism of COG through observing the activation of lymphocytes in blood. METHODS: Wister rats were used as recipients and SD rats were used as corneal graft donors, then the corneal allografts transplantation model on the closed colony rats were set up. Splenocytes proliferation and mixed lymphocyte reaction of Wister rats activated by ConA were observed. The phenotype change of CD4, CD8, CD25 in blood in different time postoperatively were observed by the di-sign flow cytometry, and the rate of CD4/CD8 was calculated. RESULTS: 1. The COG suppressed the proliferation of T lymphocytes and one-way mixed lymphocyte reaction on the corneal allografting. 2. The phenotype change of lymphocytes in boold was as follows: there was no significant difference between the different time of the CD4, CD8 expression and the CD4/CD8 rate in blood of the control group. The CD4 positive cells expressed CD25 postoperatively increased obviously. The CD4/CD8 rate of medicine group had the tendency to decrease. The CD4 positive cells expressed CD25 postoperation in the medicine group were less than that in the control group obviously. CONCLUSION: The suppression of the T lymphocyte proliferation, mixed lymphocyte reaction, CD molecule expressed by the activated T lymphocytes and the IL-2 receptor expression may be the main immunosuppressive mechanisms of Cornus officinalis glycosides on the cell-mediated immunity.

AIM: To observe the effect of ginkgolide B (GB) on glutamate-induced apoptosis in the cultured neurons of rat retina. METHODS: Neurons of rat retina were cultured and apoptosis was induced by glutamate. The neurons were cultured with different concentration of GB and the survival rate was monitored by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The apoptosis in the cultured neurons and the expression of Bcl-2 and Bax were observed by flow cytometry. RESULTS: After exposed to glutamate, the survival rate and the number of Bcl-2 positive cells obviously decreased. At the same time, the number of Bax positive cells obviously increased, and the number of the apoptotic cells also obviously increased. Such phenomena were relieved by the treatment of ginkgolide B, with raise of survival rate and the expression of Bcl-2. Meanwhile, the expression of Bax and the apoptosis of neurocytes obviously decreased. CONCLUSIONS: Ginkgolide B protects retinal neurons from the virulence induced by glutamate. Such effects of GB might be brought about by increasing the expression of Bcl-2 while decreasing Bax, resulting in increasing the ratio of Bcl-2 to Bax and so reducing the apoptosis in the cultured neurons of rat retina.

Objective To compare clinical effect of gonadotropin releasing hormone agonist(GnRH-a) alone and GnRH-a combined with low-dose dydrogesteronea and estradiol valerate on sex hormone, hypoestrogenic symptoms, quality of life and bone mineral density (BMD)in treatment of endometriosis.Methods Seventy patients with moderate or severe endometriosis, who were diagnosed by laparotomy or laparoscopic surgery within two months, were randomly assigned into two groups.35 patients in GnRH-a group were treated by goserelin (3.6 mg)for three months, and 35 patients in add-back group were treated by goserelin (3.6 mg)combined with estradiol valerate 0.5 mg and dydrogesteronea 5 mg daily.Before and after the treatment, clinical parameters were recorded and analyzed, including visual analog scale (VAS), medical outcomes survey short form 36 (SF-36), Kupperman menopausal index(KMI), BMD, the serum level of follicle stimulating hormone (FSH), estradiol (E_2) and bone gla-protein (BGP) .The first menstruation and VAS were also followed up after treatment.Results Every 3 cases in two groups lost follow-up.(1)Reproductive hormone: the level of E_2 in add-back group [(94 ± 71) pmol/L]was significantly higher than (54±52) pmol/L in GnRH-a group(P <0.01).The level of FSH in add-back group [(3.0 ± 1.9) U/L]was significantly lower than (5.7 ± 2.9) U/L in GnRH-a group (P < 0.05).(2) VAS: after treatment, VAS in both group decreased significantly when compared with that before treatment(P < 0.05), and remained until menstruated.(3) KMI: KMI in add back-group (10 ± 8) was significantly lower than (14 ± 6) in GnRH-a group (P < 0.05).(4) BMD: compared with that before treatment, BMD decreased significantly after treatment in GnRH-a group (P < 0.05), no remarkable difference of BMD was observed before and after treatment in add-back group.Before treatment, serum BGP in both groups did not show statistical difference.After treatment, the level of BGP in GnRH-a group [(7932±5206) ng/L]was significantly higher than (5419±2917) ng/L in add-back group (P <0.05).Conclusions GnRH-a combined with estrogen-progesterone regimen could relieve pain from endometriosis as effectively as GnRH-a alone and reduce hypoestrogenic symptoms and bone loss.Therefore,it is a safe and effective treatment.

Objective To evaluate effects and safety of gonadotrophin-releasing hormone agonist (GnRH-a) combined with transdermal estradiol and medroxyprogesterone acetate in the treatment of endometriosis. Methods From January I st, 2007 to July 31 st, 2007, 28 endometriosis patients underwent laparnscopic or transabdominal surgery in Obstetrics and Gynecology Hospital affiliated to Fudan University were randomly divided into group A and group B. 14 patients in group A received 3.6 mg goserelin once every 4 weeks, 12 weeks in all 14 patients in group B received goserelin and added 1/2 piece of half-hydrate estradiol every week and 6 mg oral medroxyprogesterone acetate per day, 12 weeks in all. Serum estradiol (E2 ), follicle stimulating hormone(FSH), bone gla protein levels, visual analogue scale (VAS) of pain, bone mineral density of lumbar spine, vaginal exfoliate cell spurs and the form of Kupperman were compared in patients before and after treatment. Results (1 ) After treatment, the level of FSH and E2levels were (5.0 ± 2. 6 ) U/L and (29 ± 17 ) pmol/L in group A and (3.0 ± 1.5 ) U/L, and (87 ± 53 ) pmol/L in group B, which were significantly lower than those before treatment [FSH (17. 0 ± 12. 2) U/L, and E2 (184 ± 194) pmol/L in group A and FSH :(15.3±13.6)U/L and E2: (281±242) pmol/L in group B, P ＜ 0. 01]. On the seventh day after three-month GnRH-a treatment, it was observed that the level of E2 was higher and FSH was lower in group B than the level of E2 and FSH of group A (P ＜ 0. 01 ). (2 ) After treatment, the basal vaginal exfoliate cell proportion in group A [(66. 2 ± 29. 0) %] was significantly lower than that in group B [(11.8 ± 28. 0) %, P ＜ 0. 01] ; while patients in group A owned a lower proportion of the middle [(29. 1 ± 23.1 ) %], superficial layers [(4. 0 ± 5.5 ) %] and esinophilic cells [(2. 3 ± 2. 6)%]than patients group B [middle layer: (73. 0 ± 25.2)% ; superficial layer: (15. 2 ± 10. 9)% ; esinophilic cells: (10. 8 ± 7.9 ) % ; P ＜ 0. 01]. (3) Before the treatment, patients' VAS scores of total, pelvic pain, dysmenorrheal and dyspareunia were 7.43±3. 20,2. 35 ± 1.82, 4. 93 ± 1.98 and 0. 14±0. 53 in group A and were 7.71±2. 02, 2. 57 ± 1.60, 4. 86 ± 1.56 and 0. 29 ± 1.07 in group B; after treatment, the scores above were changed to 0. 14±0. 36,0. 07±0. 27,0. 07±0. 27and 0 in group A and 0. 36±0. 50, 0. 29±0. 47, 0. 07±0. 27 and 0 in group B, which were all significantly lower than those before treatment separately (P ＜0. 01 ). When menstruation recovered, the scores were 0. 21±0. 43, 0. 07±0. 27, 0. 14 ± 0. 36, and 0 in group A and 0. 50±0. 65, 0. 29±0. 47, 0. 21±0. 43 and 0 in group B, which were also significantly lower than those before treatment (P ＜ 0.01 ), however, no statistical difference was found between groups at any time spot(P ＞ 0. 05). (4) In group A, the bone density after treatment [(0. 96 ± 0. 06 ) g/cm2] was lower than that before treatment [(0. 99 ± 0. 06 ) g/cm2, P ＜ 0.01 )]. In group B, the index was (0. 98 ± 0. 09) g/cm2, which was lower than that before treatment [(0. 99 ± 0. 10 ) g/cm2, P = 0. 201]. No statistical difference was found between groups(P ＞ 0. 05 ). The bone loss rate were (- 2. 77 ± 1.97 ) % in group A and (- 0. 93 ± 2. 86 ) % in group B (P = 0. 058 ). Before treatment, the bone gla protein was (13±3) μg/L in group A and (13±6) μg/L in group B. After treatment, the bone gla protein levels was (17±6)μg/L in group A, which was higher than that before treatment (P ＜ 0. 01 ), the level was (16±6)μg/L in group B, which was higher than that before treatment, however showed no statistical difference(P =0. 053). No difference was found in bone gla protein before and after treatment between two groups (P＞0. 05). (5) The form of Kupperman after treatment were 15±7 in group A and 11±6 in group B, which did not show significant difference (P ＞ 0. 05 ). The incidence of flash and sweat were 93% (13/14)in group A, which was significantly higher than that 57% (8/14) in group B(P ＜0.01 ). Conclusion The add-back therapy that consists of an estradiol patch and oral medroxyprogesterone acetate is effective and safe treatment for endometriosis.

Objective To investigate the antimicrobial susceptibility of spectinomycin?minocycline?azithromycin and sparfloxacin to mycoplasma(Uu and Mh)and therapeutic effect of spectinomycin to my-coplasma infection in genitourinary tract.Methods①The susceptibility test:each of the4drugs was divided into two concentrations.One was at1?g/mL(sensitive concentration)and the other was at4?g/mL(resistant concentration).If mycoplasma does not grow in both concentrations,it means the drug tested is sensitive.If it grows in both concentrations,the drug tested is resistant.If mycoplasma grows in lower concentration and does not in higher concentration,it means moderate sensitive.②Treatment regimen:Spectinomycin was injected,2g/d IM,for7-10days as a course of treatmeant.Patients were followed-up7days later and2~4weeks after treatment.Results①Among1658specimens,519were found Uu positive,and61Mh positive.The resis-tance rates of Uu to4different drugs were:7.7%for minocycline,21.4%for sparfloxacin,13.9%for azithromycin and7.3%for spectinomycin.Whereas,those of Mh were:18.0%,45.9%,54.1%,and29.5%re-spectively.②The clinical effect of spectinomycin was:out of43treated patients,37(86.0%)cured,4(9.3%)markedly improved,2(4.7%)failed.Total effective rate was95.3%and so was the elimination rate of my-coplasma.Conclusion The resistant rate of mycoplasma to spectinomycin is lower than that to minocycline?azithromycin and sparfloxacin,and the former is widely used in the treatment of mycoplasma(especially Uu)infection,with a satisfactory clinical effect.