Two lipase active fractions Lip1 and Lip2 were purified from the cell extract of Rhizopus chinensis CCTCCM201021.Both gave a single band on SDS-PAGE after using ammonium sulfate precipitation、Phenyl-Sepharose FF、DEAE-Sepharose FF and Sephadex G100 gel filtration chromatographies.The molecular masses of two lipases were 59.2kD and 39.4kD respectively.Lip1 and Lip2 showed optimal pH at 8.0 and 8.5 and their optimal temperatures were 40℃ and 35℃ respectively.The substrate specificity of the two lipases was obviously different.Lip1 was more specific to long chain fatty acid of p-nitrophenyl esters while Lip2 had a preference for the hydrolysis of short chain fatty acid of p-nitrophenyl esters.Lip1 had 1,3-position specificity for triacylglycerols hydrolysis while Lip2 had nonspecific position.Both lipases were stimulated by Ca 2+、Mg 2+ while SDS had strong inhibition on their activities.Lip1 and Lip2 had good stability in cyclohexane、hexane、heptane and isooctane(30% V/V).

OBJECTIVE:To evaluate the efficacy and safety of salmon calcitonin plus Xianling gubao for osteoporosis and ostealgia in postmenopausal women. METHODS:160 women with postmenopausal osteoporosis and ostealgia were randomized to 3 groups (treatment group and 2 control groups):the treatment group received(500 IU q.d) salmon calcitonin nasal spray plus Xianling gubao(1.5 g b.i.d orally); the control group Ⅰ received(500 IU q.d) salmon calcitonin nasal spray alone and the control group Ⅱ received Xianling gubao(1.5 g b.i.d orally) alone. All the patients received oral Caltrate D (1 tablet q.d). A course was defined as 30 days. After treatment for a total of 3 courses,the nature and degree of ostealgia in all the patients were assessed,and the outcome indexes were measured and the adverse drug reactions were recorded. RESULTS:There were significant differences between the treatment group and two control groups in total effective rates(92.59% for treatment group vs. 69.81% for control group Ⅰ and 67.92% for control group Ⅱ,P0.05). CONCLUSION:Treatment of osteoporosis and ostealgia in postmenopausal women with salmon calcitonin plus Xianling gubao has been proved to be safe and effective in that the bone density can be increased effectively and the osteoporotic pain of lumbar and back of the patients can be relieved.

Background and purpose:Although FDG tumor imaging has been applied in clinic widely, dual-phase imaging can provide much more information about the FDG uptaking of pulmonary lesions. The purpose of the study was to evaluate the usefulness of dual-phase18F-FDG coincidence detection SPECT/CT imaging in the differential diagnosis of the pulmonary lesions.Methods:There were 28 patients with pulmonary lesions which were detected by CT. All the patients undertook the SPECT/CT imaging at 2 time-phases respectively: early imaging at 40-60 min and delayed imaging at 2-3 h after the intravenous injection of FDG. Data processing: calculating the radio of T and N in early and delayed imaging respectively; T: The radioactive count of the lesions; N: The radioactive count of the normal tissue; and the change rate:ΔT/N. ROC was used to ifnd out the threshold of T1/N1, T2/N2及ΔT/N in the differential diagnosis between benign and malignant lesions. AUC was used to evaluate the diagnosis value of the dual-phase and single-phase imaging.Results:The threshold of T1/N1 in early imaging was 2.65, whereas AUC was 0.767. The sensitivity, speciifcity and accuracy were 83.3%, 30% and 64.3%, respectively. The threshold of T2/N2 in delayed imaging was 3.14, whereas AUC was 0.847. The sensitivity, speciifcity and accuracy were 94.4%, 60.0% and 82.1%, respectively. The threshold ofΔT/N in delayed imaging was 16.9%, whereas AUC is 0.950. The sensitivity, speciifcity and accuracy were 88.5%, 71.4% and 86.2%, respectively.Conclusion:Dual-phase18F-FDG coincidence detection SPECT/CT imaging has much higher accuracy and speciifcity. However it still has false positivity, and should be analyzed with CT and clinical history.

Animals ; Carcinoma, Hepatocellular ; blood supply ; pathology ; therapy ; Cell Line, Tumor ; Cells, Cultured ; Cytotoxicity, Immunologic ; immunology ; Female ; Humans ; Interleukin-12 ; pharmacology ; Interleukin-2 ; pharmacology ; Killer Cells, Natural ; cytology ; drug effects ; immunology ; Liver Neoplasms, Experimental ; blood supply ; pathology ; therapy ; Lymphocyte Activation ; drug effects ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Microcirculation ; drug effects ; Random Allocation ; Xenograft Model Antitumor Assays

Insertion of foreign genes into the genome map of Chinese herb always produces the intended beneficial effects, but also the unintended effects. Analysis of genomic function and metabolic end products based on the principle of substantial can reveal the influence on the quality of herbs, which is able to provide valuable refer-ence for the following evaluation on pharmacology and toxicology.

Objective To explore the possible effect of combined treatment of zoledronic acid and doxorubicin on proliferation,invasion and adhesion of SGC7901 cell line,and potential underlying mechanisms. Methods MTT and Transwell experiments were used respectively to investigate the effect of different concentrations of zoledronic acid and doxorubicin on the proliferation,adhesion and invasion of SGC7901. The mRNA levels of MMP2, TIMP2, MMP9, ICAM1 and CD44 were quantified by real time PCR. Results Gastric cell line SGC-7901 were incubated with zoledronic acid for 48 h in different concentration of 10-6,10-5,10-4,10-3 mol/L,their inhibition rates were 4.98％,12.19％,27.34％ and 73.13％ separately,which were higher than control group (P＜0.01),and enhanced along with the dose increasing and the time extension (P＜0.05).Gastric cell line SGC-7901 was incubated with zoledronic acid 10-4 mol /L and doxorubicin 0.5 mg/L,respectively,or in combination for 2 h,4 h,6 h,and thereafter,the detected adhesion rates were 5.03％,24.38％ and 59.65％ in control group,4.40％,19.77％ and 51.22％ in zoledronic acid group,4.28％, 18.62％ and 51.02％ in doxorubicin group,4.02 ％,15.02％ and 46.95％ in combination group. The adhesion rates were reduced after incubation with different medicament concentration (P＜0.05).In the study of cell invasion,the cell numbers per high power field were:105.11 ± 10.43,98.37 ± 15.75,96.19 ± 9.24,83.02 ± 19.72 after treatment with zoledronic acid,doxorubicin or their combination at different concentrations. Compared with control, signal medicament had weak effectiveness in invasiveness in SGC-7901 (P＞ 0.05),whereas in combination group it was significantly decreased (P＜0.05).Both zoledronic acid and doxorubicin decreased the mRNA expressions of CD44,MMP2,MMP9,ICAM1 while increased TIMP2 mRNA level (P＜0.05or P＜0.01) versus control.The effect was more evident with combination treatment(P＜0.05 or P＜0.01),compared with doxorubicin group. Conclusions Both zoledronic acid and doxorubicin can inhibit the proliferation,adhesion and invasion of SGC7901. Combination treatment has additive effect.It may be related to the down-regulation of mRNA of ICAM1,MMP2,MMP9,CD44 and up-regulation of mRNA of TIMP2.

Objective To investigate the efficacy,safety and localization of multilayer amniotic membrane transplantation(AMT) for corneal perforation associated with ulceration.Design A retrospective clinical case series.Participants Nine patients(9 eyes)with corneal perforation secondary to ulceration were enrolled into this study.These were little response to medicine,including bacterial keratitis(4 eyes ),fungal keratitis(2 eyes),fungal and bacterial mixed keratitis( 1 eye),virus keratitis( 1 eye),and Mooren's ulcer ( 1 eye).Size of perforation was 0.5～3.0 mm in diameter.Methods The AMT was applied to tamp the perforation,fill the ulcer and cover the surface of ulcer.After surgery,the medicine was continued to be used to treat the original corneal ulcer.The follow-up ranged from 6～20 months.In the suffering eyes,reformation of anterior chamber,healing of ulcer,complications and recurrence of ulceration were observed.Main Outcome Measures Of the postoperative eyes,reformation of anterior chamber,healing of ulcer,complications and recurrence of ulceration.Results The anterior chamber reformed at the first postoperative day in 9 eyes,and kept in normal depth in the follow-up time.At postoperative 2 months,the ulcer healed with sear and a smooth surface.Corneal thickness of the ulcer area recovered almost to normal.During follow up,no recurrence of ulceration or severe complications was noted.Conclusions Multilayer AMT is a safe and effective method for the management of small corneal perforation associated with ulceration,but the ulcer healed with scar.(Ophthalmol CHN,2008,17:101-103)

Objective To observe the effect of intra-articular injection of dehydroepiandrosterone (DHEA)on the experimental osteoarthritis in rabbits and study the mechanism.Methods Forty rabbits un derwent unilateral anterior cruciate ligament transection(ACLT)and then divided into two groups randomly. 100?mol/L DHEA resolved in the dimethylsulphoxide were injected into the knees of experimental rabbits 4 weeks after transection,once a week for five weeks.Rabbits in the control group were treated under the same schedule using dimethylsulphoxide.All rabbits were killed 9 weeks after ACLT and the knee joints were evalu- ated by gross morphology and histology.The mRNA expression of metalloproteinases-3(MMP-3),tissue in- hibitor of metalloproteinases-1(TIMP-1)and interleukin-lbeta(IL-1?)in the cartilage and synovium was analyzed using reverse transcription polymerase chain reaction(RT-PCR).Results Gross morphologic in- spection and histological evaluation showed that the extent and grade of cartilage and synovium damage in the experimental group were less severe than the control group.The mRNA expression of MMP-3 in cartilage and synovium decreased significantly in the experimental group(both P＜0.05).The mRNA expression of TIMP-1 in cartilage and synovium increased significantly in the experimental group compared with that in the control group(both P＜0.05).No significant difference of IL-1?mRNA expression in cartilage was found between the experimental and the control groups(P＞0.05).The mRNA expression of IL-1?in the synovium was signifi- cantly suppressed in the experimental group compared with that in the control group(P＜0.01).Conclusion DHEA protects against cartilage degradation,alleviates synovium inflammation and inhibits the progression of osteoarthritis in the experimental model.Down-regulation of MMP-3 and up-regulation of TIMP-1 in cartilage and synovium and IL-1?in the synovium may be the mechanism of the protective effect of DHEA on os- teoarthritis.

AIM: To investigate the efficacy, safety and localization of multilayer amniotic membrane transplantation (AMT) for the management of corneal perforation associated with ulceration.METHODS: Six eyes (6 patients) were treated with multilayer AMT for corneal perforation associated with ulceration. The perforation size was 0.5-2mm in diameter. The amniotic membrane (AM) was applied to tamp the perforation, fill the ulcer and cover the surface of ulcer. The follow-up ranged from 5-19 months.RESULTS: The anterior chamber reformed at the first postoperative day in all patients, and kept in normal depth in the follow- up time. The epithelialization of the AM grafts was observed in about 3 weeks after surgery. During the first postoperative month, the corneal thickness at the perforation site gradually increased, and the stromal inflammatory cell invasions surrounding the corneal ulcer were gradually subsided.Two months postoperatively, the ulcer healed with scar with part neovascutarization; the corneal thickness of the ulcer area was stable and recovered almost normal. The vision was not improved in all patients after operation. During-follow up,no severe complications or recurrence of ulceration was noted.CONCLUSION: Multilayer AMT is an effective and safe method for the management of small corneal perforation associated with ulceration, but the ulcer healed with scar.

AIM: To evaluate the efficacy and safety of annular bubble conjunctival excision combining with amniotic membrane transplantation(AMT) for vernal keratoconjunctivitis(VKC) with gelatinous swellings(papillae) at the limbus.METHODS: Twenty eyes(10 patients) with VKC, were characterized by gelatinous swellings(papillae) at limbus and were underwent annular bubble conjunctival excision associated with AMT. The follow up ranged 5-21months.RESULTS: After operation, the symptoms were subsided remarkably: itching,discharge and foreign body sensation were almost disappear; photophobia and tearing were obvious in the first week after surgery, but were gradually alleviated, and almost disappear after one month of postoperative.Yellow-brown or filthy red gelatinous protuberances at limbus were cleaned away by operation and did not recur during the follow up. Corneal superficial punctuate keratitis(11 eyes) and corneal ulcers(3 eyes) were healed during the first week after surgery and did not recur. Conjunctival congestion was gradually lessened after surgery, which could not be noted 1months after surgery in 19 eyes, and the conjunctiva around the limbus, where AMT was performed, was whiter than the normal. But conjunctival congestion in 1 eye lasted for about 5mon accompanying with a little of itch, and this eye was treated with eyedrops with improvement but recurrence. No severe complications had been seen during the follow up.CONCLUSION: Annular bubble conjunctival excision combining with amniotic membrane transplantation is an effective and safe method for the treatment of vernal keratoconjunctivitis with gelatinous swellings (papillae) at limbus. But the candidates must be chosen with discretion.