BACKGROUND:Host immune rejection is the main problem for nerve alograft in the repair of nerve defects. Therefore, how to avoid and minimize the immune rejection is the key to the success of nerve alografting. OBJECTIVE: To develop a new nerve pretreatment method by which Schwann cels and myelin can be removed from the peripheral nerve of dogs while the basilar membrane can be reserved integraly in order to obtain acelular nerve alografts. METHODS:Bilateral sciatic nerves from healthy adult dogs were taken and pretreated with the combined freeze-thaw and chemical methods folowed by microscopic observation of ultrastructural features, histological staining and western blot analysis of its ingredients. RESULTS AND CONCLUSION:Pretreated acelular nerves with good ductility and excelent epineurium toughness were empty basal lamina tubes with no Schwann cels, myelin and fragments that were al removed thoroughly, but the basilar membrane was fuly retained. These findings indicate that the optimized combination of freeze-thaw and chemical methods can efficiently clear Schwann cels and myelin which are the major antigenic components in the peripheral nerve, while preserve the basilar membrane to promote nerve regeneration. Therefore, this method can be an ideal method for preparation of tissue engineered nerves.

Three hybridomas producing monoclonal antibodies (Mab) to zona pellucida of porcine ovarian egg(ZP-OVA) were established(LPDg, LPC4, LPD,).Two Mab(LPD_8 LPC_4) were examined against human ovarian tissue by immunohistochemical technique in vitro. We found that the two Mab had cross-reaction with zona pellucida and ovum cytoplasm and no cross-reaction with membrana granulosa and theca folliculi. It showed that there were affinities of LPD_8 and LPC_4 to antigenic determinamts in the ovum cytoplasm and common antigenic components in zona pellucida and ovum cytoplasm. It suggested that the zona pellucida was derived from oocyte.

Objective To discuss the curative effects and side-effects of combining low-dose domestic interferon(sainuojin)with CHOPP-regimen in treating skin T-lymphoma.Method During the period June 1996 to June 2008,46 cases patient with skin T-lymphoma were treated by low-dose domestic interferon(sainuojin)and CHOPP-regimen,and all patient were received four cycle chemotherapy at least.And the usage was as follows:Sainuojin 3?106U/d,im,d1～21;CTX 600mg/m2,iv,d1;ADM 50mg/m2,iv,d1;VCR 1.4mg/m2,iv,d1;Predinisone 90 mg/d,PO,d1～5;PYM 8mg/d,im,d1,3,5,and every 3 weeks repeat.At the same time,the place of skin lesion was daubed with hydrochloric acid nitrogen mustard once everyday.Results CR 23 cases,PR 16 cases,NC 7 cases.The effective rate was 84.78%.The maximum time of survivorship was 11.6 years;the average survivorship time was 8.9?0.56 years;and the mostly side-effects were arrest of bone marrow and fever.Conclusion The curative effects of combining low-dose domestic interferon(sainuojin)with CHOPP-regimen in treating skin T-lymphoma were confirmed,and the side-effects were small.It is good for clinical popularization and application.

Objective To evaluate the clinical efficacy of triangle flap from the nasal vestibule on correcting the minor form unilateral incomplete cleft lip.Methods 72 patients with the minor-form unilateral incomplete cleft lip were invloved in this study.Based on different surgical procedures,all the patients were divided into two groups:36 patients were treated with harvesting a triangle flap from the nasal vestibule,rotating,increasing lip height;other 36 patients were treated with Millard method as control group.The positive photographs of two groups of patients were taken one year after surgery.Lip height,lip width,nostril width,nostril circumference and visible scar area were measured and compared statistically.Results Good rate of the group of the triangle flap from nasal vestibule was 91.6％ (33/36),but that of the group of Millard method was 72.2％ (29/36) (P＜0.05).The ratios of unaffected to affected sides of lip height,nostril circumference and nasal width in the triangle flap from nasal vestibule were 1.077±0.015,1.083±0.005,and 1.083±0.005;those of other group of Millard method were 1.078±0.013,1.095±0.005 and 1.096±0.015,respectively,with no significant difference (P＞0.05).But there was significant difference in laberal scar between the group of the triangle flap from nasal vestibule (0.510±0.004) mm2 and the control group of Millard method (0.830±0.009) mm2 (P＜0.05).Conclusions The nasal vestibule triangle flap method can significantly decrease the visible scar on lip and achieve the same good result compared to traditional Millard method.

Objective To compare the effects of three kinds of Oncomelania hupensis RNA extraction methods,namely a modified SDS method,TRIzol reagent method,and CTAB method,so as to obtain an economical and efficient method for RNA extraction from O. hupensis. Methods The modified SDS method,TRIzol reagent method and CTAB method were applied to ex-tract the RNA from O. hupensis. A nucleic acid protein analyzer was used to measure the concentration and purity of RNA. The yields were calculated by the concentration of the products. The purity was indicated by A260/A280 and A260/A230. The quality of RNA was inspected by 1% agarose gel electrophoresis. The β-acting gene was selected as the target gene for RT-PCR analysis. Re-sults The RNA yields obtained by using the three kinds of extraction methods were significantly different(F = 16895.85,P <0.01)according to the analysis of variance. The LSD test showed that the yields obtained by using the modified SDS method were the highest,and those obtained by the CTAB method were the lowest. The purity of RNA extracted by the CTAB method was su-perior to that by the other two methods,and the A260/A280 and A260/A230 ratios of the CTAB method were in the range from 1.8-2.0 and 2.0-2.2. The A260/A230 ratios of the other two methods were both lower than 2.0. The RNA extracted by the modified SDS meth-od had the better integrity. The electrophoresis results showed that the 28S rRNA band,18S rRNA band and 5S rRNA band were clear,and there was no obvious smear between each band. The RNA obtained by the TRIzol reagent method had no 28S rRNA band,and that obtained by the CTAB method had no 28S rRNA and 5S rRNA bands. The β-acting gene of the RNA ex-tracted by all the three methods could be amplified by RT-PCR. The costs and time-consuming of the modified SDS method were less than those of the other two methods. Conclusion The modified SDS method is an economic and efficient method,and it is suitable for extracting the RNA of O. hupensis,especially for large sample preparation.

OBJECTIVETo evaluate Amifostin's effect on protecting kidney from cisplatinum (DDP) injury and its adverse reactions and safety.

METHODS193 Patients were divided into two groups randomly: 102 in group A (treatment group) and 91 in group B (control group). Indexes such as blood routine, blood calcium, liver function, blood urea nitrogen (BUN), cretinine (C), and urinary N-acetyl-beta-D-glucosaminidase (NAG)/C and micro-albumin (MAB/C) were monitored at different intervals before or after treatment.

RESULTSIn the two courses of treatment in both groups, the deviation (D) values of MAB/C before treatment and on D2 in group A were lower than those in grop B (P < 0.05), so were those before treatment and on D4, D6, D10 and D14 (P < 0.01). The D-values of NAG/C before treatment and on D4, D6, D10 and D14 in the first course of group A were obviously lower than those on the corresponding days in group B (P < 0.01), so were those before treatment and on D2, D4, D6, D10 and D14 in the second course (P < 0.01).

CONCLUSIONThe reduction of MAB/C and NAG/C by Amifostin in group A demonstrates that: Amifostin is able to effectively protect the renal function, regardless of the type of tumor. In contrast with group B, Amifostin in group A shows no protection for tumor in lung cancer and ovarian cancer. The main side effects of Amifostin are mild hypotension, nausea, vomiting and hypocalcemia in some patients.

Adult ; Aged ; Amifostine ; adverse effects ; therapeutic use ; Antineoplastic Agents ; adverse effects ; Cisplatin ; adverse effects ; Humans ; Kidney Diseases ; chemically induced ; prevention & control ; Middle Aged ; Protective Agents ; adverse effects ; therapeutic use

This study was aimed to examine the expression of apoptosis-associated gene Fas in HeLa cell, explore the effects of the co-immobilized cytokines (tumor necrosis factor-alpha and interferon-gamma), and probe the potential mechanism of action. The preparation and application of the research couple IFN-gamma and TNF-alpha to the polystyrene cell culture plate were performed using the Photo-immobilization method, with different doses (20 ng/well and 200 ng/well) and synthesized optical active material. HeLa cells were treated with cytokines for two dose and 1, 3, 6 days. The result showed that the free cytokines induced HeLa apoptosis quickly, yet the HeLa apoptosis induced by co-immobilized cytokines had longer effect.
Apoptosis ; drug effects ; genetics ; Drug Synergism ; HeLa Cells ; Humans ; Immobilized Proteins ; chemistry ; pharmacology ; Interferon-gamma ; chemistry ; pharmacology ; Tumor Necrosis Factor-alpha ; chemistry ; pharmacology ; Up-Regulation ; fas Receptor ; metabolism

OBJECTIVE:To establish a method for the determination of brucine concentration in plasma of rats,and to compare the pharmacokinetic differences between brucine and its nanostructure lipid carrier (NLC) in rats. METHODS:Sixteen male SD rats were randomly divided into brucine NLC solution group and brucine solution group(using normal saline as solvent, and containing brucine 1.28 mg/mL),with 8 rats in each group. They were given relevant solution 10 mg/kg via tail vein. Blood sample 0.5 mL was collected from fundus venous plexus capillary before medication and 15,20,30,40,45,60,90,120,150, 180,210,240,480 min after medication. HPLC method was adopted. The determination was performed on Dikma C18column with mobile phase consisted of methanol-water containing acetic acid and triethylamine(30∶70,V/V)at the flow rate of 1 mL/min. The detection wavelength was set at 265 nm,and column temperature was 30 ℃. Sample size was 10 μ L. Pharmacokinetic parameters of rats in 2 groups were calculated by using DAS 2.0 software,and the difference of them were compared by F test. RESULTS:The linear range of brucine plasma concentration were 1.03-66.00 μg/mL(R2＝0.999 6);the limit of quantitation was 1.03 μg/mL,and lowest detection limit was 0.515 μg/mL. RSDs of intra-day and inter-day were lower than 5%;method recoveries were 84.90%-100.88%, extraction recoveries were 80.60%-91.98%(all RSDs were lower than 10%). Average plasma concentration-time curve of single administration of brucine NLC solution and brucine solution were all in line with two-compartment model after medication via tail vein. The pharmacokinetic parameters included t1/2αwere(0.24±0.11)and(0.06± 0.03)h;t1/2 βwere (2.90 ± 0.22) and (0.57 ± 0.32)h;AUC0-twere (88.00 ± 6.98) and (28.50 ± 5.87)μg·h/mL;AUC0-∞were (109.96±7.99)and(45.06±6.66)μg·h/mL. Compared with brucine solution group,t1/2 α,t1/2 β,AUC0-tand AUC0- ∞of brucine NLC solution group were increased significantly;while CL, k10and k12were decreased significantly, with statistical significance (P＜0.05 or P＜0.01). There was no statistical significance in k21between 2 groups (P＞0.05). CONCLUSIONS: Established HPLC method is simple, specific,sensitive,precise and highly recoverable. It can be used for the determination of plasma concentration and phamacokinetic study of brucine in rats. After brucine NLC is prepared,the pharmacokinetic parameters of brucine change significantly;retention time of brucine is significantly prolonged and the clearance rate decreases significantly.