2003年 · 第116卷第08期

Objective To determine the effects of raloxifene hydrochloride(RLX)on bone mineral density(BMD), bone metabolism markers and serum lipids in healthy postmenopausal women in Beijing. Methods A multicenter, randomized, double-blind, placebo-controlled study was conducted in a total of 204 healthy postmenopausal women(age 59. 5 ± 5. 0 years and weight 62. 8 ± 8. 7 kg)treated with either RLX 60 mg(n = 102)or placebo(n = 102)daily for 12 months. BMD, serum lipids, and bone markers were measured before and after drug administration. Results Compared with placebo, RLX produced a significant increase in both total lumbar spine and total hip BMD. For the lumbar spine, percentage increase in total BMD was 2. 3% with RLX compared with a decrease of 0. 1% with placebo(P < 0. 001). Corresponding values for total hip BMD were a 2. 5%increase for RLX and a 1.1 % increase for placebo(P = 0. 011). For biochemical markers of bone metabolism, serum osteocalcin and C-telopeptide, percentage decreases were 27. 65% and 24. 02% in RLX-treated subjects. Corresponding values in placebo were a 10. 64% decrease and a 15. 75% increase(RLX compared with placebo, both P < 0. 001). For total cholesterol and low-density lipoprotein cholesterol levels, percentage decreases were 6.44% and 34. 58% in the RLX-treated group. Corresponding values in placebo-treated patients were a 1. 44% increase and a 19. 07% decrease(RLX compared with placebo, both P < 0. 001). No differences were found for high-density lipoprotein cholesterol or triglyceride levels between the two groups. Only 5 subjects discontinued early owing to an adverse event(3 in the RLX group and 2 in the placebo group). Conclusions This study confirms that RLX exerts positive effects on the skeleton, increasing BMD and decreasing biochemical markers of bone metabolism, and has a positive effect on the overall serum lipid profile in postmenopausal women in China. Chin Med J 2003; 116(8): 1127-1133

Objective To evaluate the efficacy and safety of leflunomide in comparison with methotrexate(MTX)on patients with rheumatoid arthritis(RA)in China. Methods Five hundred and sixty-six patients with active rheumatoid arthritis were randomly assigned to receive leflunomide at 20 mg once daily or MTX at 15 mg once weekly in a controlled trial. Five hundred and four patients completed the 12-week treatment and some patients continued the treatment for 24 weeks. Results Both leflunomide and MTX could improve the symptoms, signs, and joint function, but there were no changes in X-ray observations of patients with rheumatoid arthritis. In the leflunomide group, the overall rates of effectiveness at 12 weeks and 24 weeks were 86. 94% and 92. 31 % respectively; the rates of remarkable improvement were 64. 95% and 79. 81% respectively. In the MTX group, the overall rates of effectiveness at 12 weeks and 24 weeks were 84. 04% and 83. 15% respectively; the rates of remarkable improvement were 56. 81 % and 75. 28 % respectively. According to intent-to-treat analysis, the ACR 20%response rates at 12 weeks and 24 weeks in the leflunomide group were 62. 54% and 67. 18% respectively, compared with 60. 08% and 61.32% respectively in MTX group. No statistical differences were shown in the efficacy between the two groups(P>0. 05). The adverse events in the leflunomide group were gastrointestinal symptoms, skin rash, alopecia, nervous system symptoms, decreased leukocyte count, and elevation of alanine aminotransferase(ALT). Most of these side effects were mild and transient. The incidence of adverse events in the leflunomide group was 16. 84%, significantly lower than that in MTX group(28. 17%, P = 0. 002). Conclusions Leflunomide is effective in the treatment of RA with less adverse events than MTX. Its efficacy is similar to MTX, but the incidence of adverse events and the rate of withdrawal due to adverse events were lower in the leflunomide group than in MTX group. Chin Med J 2003; 116(8):

Objective To assess the effects of hormone replacement therapy(HRT)on platelet activation in postmenopausal women compared with premenopausal women. Methods The expressions of CD41 and CD62P in fifteen postmenopausal women before and after HRT were detected using flow cytometry(FCM), with fifteen premenopausal women with a mean age of 47 years as controls. Results The expressions of CD41 and CD62P in postmenopausal women were higher than those in the control group. CD62P(%), CD62P(I)and CD41 were reduced from 36. 40 ± 5. 9, 37. 75 ± 5. 8, and 470. 11 ±74. 0 to 27. 97±5. 6, 26. 64±4. 9, and 303. 23±72. 8 after six months of HRT(P<0. 05). Conclusions Platelet activation in postmenopausal women was higher than in premenopausal women and was reduced significantly after six months of HRT. HRT may have a favorable effect on reduction of platelet activity. Chin Med J 2003; 116(8): 1134-1136

Objective To develop a simple, cheap, quick, accurate and practical method for a high throughout genotypes assay of human papillomavirus(HPV)DNA. Methods Crude DNA was extracted by a simplified proteinase K digesting method. HPV common conservative primers: GP5 +/6 + system was used to amplify HPV DNA in 127 samples of condylomata acuminatum(CA)and cervical scrapes by PCR, then the PCR product was assayed using a template directing terminator incorporation(TDI)and genotypes were detected with fluorescence polarization(FP). Major HPVs type-specific probes(HPV6, 11, 16, 18, 31, 33, 35 and 58)designed by us were hybridized with the specific PCR products and a special fluorescent ddNTP terminator was directly added to the end of the probe under direction of specific PCR products. The results were measured with FP and compared with the results of the DNA sequence. Results Compared with the results of DNA sequencing, the results detected with fluorescence polarization were all correct. The proposed method could detect more than one type of HPV infection, but DNA sequencing method could not. The positive rate of HPV was 100% in 78 CA biopsies. Among them, there were 14 HPV double infections [HPV6B and 11(9 cases), HPV11 and 16(4), HPV11 and 18(1)], 5 HPV triple infections [HPV6B, 11 and 16(4), HPV11, 16 and 18(1)], and one HPV quadruple infection(HPV6B, 11, 16 and 18). The positive rate of HPV was 77% in the 49 cervical scrapes. Six HPV double infections [HPV6B and 11(2), HPV11 and 16(1), HPV6B and 16(1), HPV16 and 18(1), HPV18 and 58(1)], 3 HPV triple infections [HPV6B, 11 and 16(2), HPV11, 16 and 18(1)] and one HPV quadruple infection(HPV6B, 11, 16 and 18)were detected in cervical cancer scrapes. Conclusions The proposed method allowed a high throughout, special, simple, rapid, automatic and economical detection of HPV-DNA genotyping without a use of labeling probes. It can detect multiple HPV genotype infection and will be and useful tool in HPV genotype screening. Chin Med J 2003; 116(8): 1137-1140

Objective To explore the survival time, pathological change and liver regeneration in different kinds of reduced-size liver transplantation in rats using steatotic grafts. Methods Macrovesicular and microvesicular steatotic rat liver models were established by feeding rats with a diet consisting of 79% standard chow, 20% lard and 1 % cholesterol for different time periods. With modified two cuff vascular anastomoses and end-to-end sutures on the bile duct, reduced-size orthotopic rat liver transplantations were performed in an attempt to explore the ratio of graft weight to recipient body weight, recipient original liver weight and histological and electron-microscopic findings in comparison with whole rat liver transplantations. Results A one-week survival rates for the rats undergoing whole liver transplantation, and those in the 70% reduced-size orthotopic liver transplantation(ROLT)group, the 60%ROLT group and the 50%ROLT group(grade I macrosteatotic grafts)were 91. 67%, 75%, 75% and 25%. A 2-week survival rate was 83. 33%, 75%, 58. 33% and 0 respectively. And their graft recipient body weight(GRBW)values SD were 3. 56% ±0. 36%, 2. 53% ± 0. 15%, 2. 28% ±0. 12% and 1. 83% ±0. 16%, respectively. In grade II macrosteatotic grafts, the one-week survival rate for those undergoiong whole liver transplantation and those in the 70% ROLT group was 83. 33% and 25%. In the microsteatosis grafts for whole liver transplantation, 70% ROLT, 60% ROLT and 50% ROLT, the one-week survival rate was 83. 33%, 75%, 75 % and 33. 33 %; and the 2-week survival rate was 75 %, 66. 67 %, 66. 67 % and 0, respectively. The survival rate of the 50% ROLT group using grade I macrosteatotic grafts or grafts mainly with microsteatosis was significantly different from that of other groups. While using macrosteatotic grafts in grade Ⅱ, the 1-week survival rate of the 70% ROLT group was very poor. Pathological findings after operation included liver regeneration and portal space with mild lymphocyte infiltration. Improvement in steatosis and dilation of the central vein and sinusoids was observed in some rats. Conclusions In the successful and long-term survival of rat reduced-size liver transplantation using grade I macrosteatotic grafts or grafts with microsteatosis, the GRBW values should be over 2. 28% ±0. 12%, and the value of graft-recipient liver weight should be over 60%. Steatotic livers in grade Ⅱ should not be used as grafts in ROLT. Steatosis was improved and even totally cured in some long-term survival rats. Chin Med J 2003; 116(8): 1141-1145

Objective To determine the role of the NF-kappaB/I-kappaB pathway during ischemia reperfusion(l/R)injury to rat liver. Methords A group of rats underwent partial hepatic ischemia and reperfusion. The left and median lobe of the liver was subjected to ischemia for 90 minutes followed by reperfusion for previously specified periods. NF-kappaB activity was analyzed by electrophoretic mobility shift assay(EMSA). The protein level of I-kappaB was assessed using Western blot analysis. A semiquantitative reverse transcriptase polymerase chain reaction was used to analyze TNF-α and ICAM-1 mRNA levels. Results During liver l/R injury, NF-kappaB activation was induced in a time-dependent fashion. NF-kappaB was activated within 1 hour and 2 hours after the initiation of reperfusion and decreased afer 4 hours. The I-kappaB protein level was decreased in the cytoplasm after 2 hours, and the messenger RNA expression of TNF-α and ICAM-1 were increased simultaneously. Conclusions The data suggest that I-kappaB protein was degraded during hepatic l/R injury, NF kappaB was realeased and bound to special sequence in the promoters of budget genes, which regulated the expression of TNF-α and ICAM-1 mRNA. This provides evidence that the NF-kappaB/I-kappaB pathway plays an important modulating role in the expression of proinflammatory genes relevant to the development of ischemia reperfusion injury. Chin Med J 2003; 116(8): 1146-1149

Objective To investigate the synergistic antitumor effects of combined use of p14ARF gene and 5-fluorouracil(5-Fu)in pancreatic cancer. Methods A human pancreatic cancer cell line PC-3 was transfected with lipofectin-mediated recombinant p14ARF gene, and was then administered with 5-Fu. Cell growth, morphological changes, cell cycle, apoptosis, and molecular changes were measured using the MTT assay, flow cytometry, RT-PCR, Western blotting, and immunocytochemical assays. Results After transfection of p14ARF, cell growth was obviously inhibited, resulting in an accumulation of cells in the G1 phase. The proportion of cells in the G1 phase was significantly increased from 58. 51 % to 75. 92 %, and in the S and G2/M phases decreased significantly from 20. 05% to 12.60%, and from 21. 44% to 11.48 %, respectively, as compared with those of the control groups. PC-3/p14ARF cells that underwent 5-Fu treatment had significantly greater G2/M phase accumulation, from 11. 48% to 53. 47 %. The apoptopic index was increased in PC-3/p14ARF cells from 3. 64% to 19. 62%. The MTT assay showed p14ARF-expressing cells were significantly more sensitive to 5-Fu(0. 01-10 mg/L)than those devoid of p14ARF expression(P<0. 01). Western blotting showed p14ARF upregulates p53 expression. Conclusion Combined use of p14ARF gene and 5-Fu acts synergistically to inhibit pancreatic cancer cell proliferation, suggesting a new anticancer strategy. Chin Med J 2003; 116(8): 1150-1155

Objective To identify genetic abnormalities in primary pancreatic carcinoma in humans. Methods Comparative genomic hybridization(CGH)was used to investigate genomic imbalances in 27 cases of pancreatic carcinomas. Multiple deletions and gains were observed in all tumor specimens. Results Losses affecting chromosomes 9p, 17p, 4q and 6p and gains involving 8q, 7q, 3q and 1 q were commonly observed. Conclusions There are multiple regions of chromosomes with changes copy number in pancreatic carcinoma. The altered chromosomal regions may contain several candidate genes which are involved in the development and progress of pancreatic carcinogenesis. Chin Med J 2003; 116(8): 1156-1160

Objective To study the effect of xenotransplantation with pig parathyroid cells, which was prepared using cell microencapsulation technique, on the treatment of hypoparathyroidism in rats without immunosuppressor. Methods Parathyroid cells were isolated from 10 healthy newborn pigs and encapsulated in alginate-polylysine-alginate(APA)membranes. Thirty-two aparathyroid Wistar rats were randomly allocated to microcapsule, non-microcapsule, empty microcapsule, and control groups. Each rat was injected intraperitoneally with encapsulated porcine parathyroid cells, free porcine parathyroid cells, empty capsules or 0. 9% NaCI, respectively. Total serum calcium and parathyroid hormone levels were monitored continuously for 40 weeks. And then, the transplant beds were retrieved and subjected to morphologic and eloctron microscopic examination. Results In those animals xenotransplanted with microencapsulated porcine parathyroid cells, the calcium and PTH levels were consistently within the normal range during the 40 weeks. In contrast, no therapeutic effects were observed in rats in the non-microcapsule group. Furthermore, neither empty capsules nor 0. 9% NaCI were shown to have any effect on the recipient' s serum calcium or PTH levels. After 40 weeks, electron microscopic examination demonstrated that the parathyroid cells within the microcapsules had survived well in vivo. Conclusions Xenotransplantation of microencapsulated newborn pig parathyroid cells can successfully treat hypoparathyroidism in rats without using immunosuppressive drugs. The results of this study show the possible clinical use of microencapsulated porcine parathyroid cells. Chin Med J 2003; 116(8): 1161-1165

Objective To explore the therapeutic alternatives and evaluate the related clinical results of patients with primary central nervous system lymphoma(PCNSL)treated with gamma knife radiosurgery(GKS). Methods From January 1995 to December 2001, 44 patients suffering from PCNSL, who had undergone stereotactic biopsy or craniotomy, and who had received a confirmed dignosis through pathological examination, were treated with GKS. All cases were followed up for 1-46 months with an average postoperative period of 27 months. The clinical materials, image features, treatment methods and results of follow-up, were retrospectively reviewed. Results The symptoms and signs of the patients were markedly improved within 1-3 weeks after GKS. The Kanofsky performance status was also improved from a preoperative average of 40% to a postoperative one of 90%. Thiry-eight patients(86. 36%)were in complete remission(CR), the other six(13. 63 %)were in partial remission(PR). The local control rate reached 100 %, and the median survival time was 26. 5 months. The main side effect was brain edema, which can be treated with dexamethasone and mannitol. Conclusion GKS is a safe and effective method in multimodality treatment of PCNSL. A stereotactic biopsy coupled with GKS is the first choice for diagnosis and treatment. Adjuvant chemotherapy or radiotherapy should then be given according to the patient's condition. Chin Med J 2003; 116(8): 1166-1170

Objective To explore digestive system manifestations in patients with severe acute respiratory syndrome(SARS). Method The clinical data of 96 cases with SARS admitted into our hospital from February 6, 2003 to March 28, 2003 were retrospectively analyzed. Results Among the 96 cases, 26 cases(27%)had diarrhea, 17(18%)had nausea, 6(6%)had vomiting, 16(17%)had bellyache, and 8(8%)had ALT elevation. Conclusions Patients with SARS may have digestive system manifestations; diarrhea is the most common symptom. Chin Med J 2003; 116(8): 1265-1266

Objective This study was conducted to investigate the function of the first intron of porcine growth hormone(pGH)gene in the gene expression. Methods PCR method was used to amplify the first intron from pig genomic DNA. The intron was then inserted into pGH cDNA to construct pGH cDNA-intron(pGH cDNA-in). The recombinant adenoviruses containing pGH cDNA and pGH cDNA-in genes under control of CMV promoter were generated by homologous recombination method in HEK 293 cells respectively. The effect of the first intron on gene expression was evaluated by comparing the expression levels of pGH cDNA-in and pGH cDNA mediated by adenovirus vectors in CHO cells. Results The expression level of pGH cDNA containing the first intron increased by 117%, which was significantly higher than that of pGH cDNA without the intron(P<0. 001). Conclusion The first intron of pGH gene has the function to improve pGH gene expression. Chin Med J 2003;116(8): 1267-1269

Objective To study the gene expression profiles of human bone marrow derived mesenchymal stem cells and tendon cells. Methods Total RNA extracted from human bone marrow derived mesenchymal stem cells and tendon cells underwent reverse transcription, and the products were labeled with α-32 P dCTP. The cDNA probes of total RNA were hybridized to cDNA microarray with 1176 genes, and then the signals were analyzed by Atlaslmage analysis software Version 1. 01a. Results Fifteen genes associated with cell proliferation and signal transduction were up-regulated, and one gene that takes part in cell-to-cell adhesion was down-regulated in tendon cells. Conclusion The 15 up-regulated and one down-regulated genes may be beneficial to the orientational differentiation of mesenchymal stem cells into tendon cells. Chin Med J 2003; 116(8): 1270-1272

Objective To investigate pre-and postoperative changes of regional cerebral cortical blood flow in patients with cerebral arteriovenous malformation. Method Twenty-two adult patients with arteriovenous malformation(AVM)were recruited into this study at Beijing Tiantan Hospital from September 2001 to May 2002. Eight patients had giant cerebral AVM and the other 14 had a small one. Cortical cerebral blood flow(CBF)was measured by laser Doppler flowmetry(LDF)before and after AVM resections. After surgery, the probe of LDF was implanted adjacent to the area of AVM and monitored for 24 hours. Results CBF increased significantly after the resection in all patients regardless of AVM size. In patients with small AVM, CBF returned to the baseline level within 4 hours, but in patients with giant AVM, CBF remained high even after 24 hours. Conclusions Monitoring CBF is helpful to understand pre-and postoperative changes of regional cortical CBF in patients with cerebral AVM. Chin Med J 2003; 116:(8)1273-1275