2002年 · 第115卷第05期

Objective To investigate the effect of rehabilitation therapy for short bowel syndrome on patient nutritional status and intestinal adaptation. Methods The rehabilitation therapy included enteral or parenteral nutrition, glutamine, recombinant human growth hormone and rehabilitative diet. From January 1997 to July 2000, twenty-seven patients with short bowel syndrome received the treatment. The average age of the patients was 38.5 ± 19.3 years, and the length of residual small intestine ranged from 15 to 80 cm, with an average of 46.8 ± 23.4 cm. The ileocecal valve was preserved in 14 cases, and the average time between the onset of short bowel syndrome and the rehabilitation therapy was 86 ± 105 days. Results After the treatment, nutritional status of the patients improved markedly, and intestinal absorptive capacity improved. Eight patients were followed up for more than 2 years, among whom 4 (50%) were weaned from total parenteral nutrition . Thirteen patients were followed up for more than 1 year, and 10 patients (76.9%) were weaned from total parenteral nutrition. Conclusions Rehabilitation therapy for short bowel syndrome can improve patient nutritional status effectively and promote intestinal adaptation, providing a new hope for these patients. The therapeutic effects are related to the length of the residual small intestine, patient's age and duration between massive intestinal resection and start of the treatment. Early initiation of rehabilitation therapy promotes intestinal adaptation and increases patient's ability to wean from total parenteral nutrition. Chin Med J 2002; 115(5): 776-778

Objective To understand the distribution of prevalence rate of osteoporosis in the middle-aged and elderly in parts of China. Methods Bone mineral density (BMD) was measured and questionnaires were taken for 5593 people aged above 40 years in five administrative areas in China selected by the stratified-multi-steps-cluster sampling method. Results The total prevalence rate of osteoporosis was 16.1%. The prevalence rate among males was 11.5%and among females was 19.9%(P<0.01). There were also osteoporosis prevalence differences among cities, age groups, gender groups and areas. Conclusion The prevention and treatment of osteoporosis are very important for females but also should not be ignored in males. Chin Med J 2002; 115(5): 773-775

Objective To investigate whether transdermal scopolamine increased cardiac vagal activity in patients during the acute phase of myocardial infarction. Methods 30 patients with a first acute myocardial infarction and preserved sinus rhythm who were on no drug that could influence the sinus node were randomly assigned to either treatment group or placebo group. Measures of heart rate variability (HRV) in patients given drug or placebo were obtained by digital 24 hour Holter recording before and after treatment. Baroreflex sensitivity was performed using the phenylephrine method. Results No significant differences was found in the indices of the time domain and the frequency domain in both groups before treatment. Patients with transdermal scopolamine showed a significant increase in the standard deviation of normal RR intervals (SDNN), standard deviation of all five min mean normal RR intervals (SDANN), root mean square of differences of successive normal RR intervals (rMSSD), total power (TP, 0.00-0.40 Hz), low frequency peak (LF, 0.04-0.15 Hz), high frequency peak (HF, 0.15 -0.40 Hz), and Baroreflex sensitivity after treatment (P <0.05 - 0.01) . These indices did not change in patients given placebo. Conclusion Low doses of transdermal scopolamine safely increase cardiac parasympathetic activity and improve autonomic indices in patients with acute myocardial infarction. Chin Med J 2002; 115(5): 770-772

Objective To investigate the proliferation and differentiation of cultured corneal stem cells and determine the effect of corneal stem cells cultured on amniotic membranes on the limbal area for treating corneal burns. Methods The proliferation and differentiation of corneal stem cells in vitro had been examined using colony-forming efficiency and immunohistochemistry. The stem cells had been cultured on amniotic membranes and transplanted to the limbal area for treating corneal burns. Results Corneal stem cells had a high proliferation capacity in primary and first passage, cytokeratin 3 was not expressed in primary culture but partly in first passage. The stem cells could proliferate to form cell layer on an amniotic membrane. When transplanted, stem cells could survive on limbus. After transplantation, ocular inflammation resolved, the cornea re-epithelialized, the stromal opacity reduced, the superficial neovascularity was lessened and the conjunctival fornix re-established. Conclusions Ocular surface conditions could be improved by allograft of corneal stem cells cultured on amniotic membranes. Chin Med J 2002; 115(5): 767-769

Objective To investigate the intraspecific difference of the triose phosphate isomerase (tim) gene from Giardia lamblia (G . lamblia) . Methods Total genomic DNA of G. lamblia was extracted and partial fragments of the triose phosphate isomerase (tim) gene were amplified by polymerase chain reaction (PGR). All nucleotide sequences were analyzed by using a phylogenetic analysis, which was constructed with parsimony and Neighbor-joining (NJ) methods. Results A total of 124 variable sites (23%of all sequences detected) was defined, most of which were found at the silent sites of codons. Two similar phylogenetic trees were constructed, subdividing 16 Giardia isolates into two groups. Conclusion The genetic diversity of G. lamblia appeared to be little affected by factors of both host and geography, while natural-selection played an important role in DNA molecular evolution level of the tim gene. The tim gene may be considered a very useful genetic marker of the population genetic structure of G. lamblia. Chin Med J 2002; 115(5): 763-766

Objective To evaluate the anti-hepatoma effect of Calmodulin antagonist 0-4-ethoxyl-butyl-Berbamine(EBB), one of the berbamine derivatives. Methods Monotetrazolium (MTT) method was used to analysize the effect of EBB on the proliferation and growth inhibition effect. Of a hepatoma cell line in vitro. A mouse hepatoma model was induced by injection of hepatoma ells (H22) in the abdominal cavity. The effect of EBB on survival at different concentrations as well as in combination with 5-FU were investigated in vivo. Flow cytometry analysis, dot blot hybridization,western blot, immunochemistry, enzyme-linked lectin assay (ELISA), trifluoperazine (TFP) and electron microscopic observation were used to study the effect of EBB on cell cycle process, P53 mRNA and protein levels, calmodulin content and ultrastractural changes of hepatome cells. Results EBB exerts a very strong inhibitory effect on human hepatoma cell line 7402 and mouse hepatoma cell line H22 in vitro. The IC50 value of EBB for the two cell lines are 3.312 μg/ml and 1.167 μg/ml, respectively. The sensitivity of H22 cells to 5-FU can be markedly enhanced: The IC50 dosage of 5-Fu can be decreased from 0.75 μg/ml down to 0.15 μg/ml, when jointly administered with nontoxic dosages of EBB (IC10) . In vivo, EBB can prolong the lifespan of mice with ascites H22 to more than three months. 64%of mice survived, while all animals in the control group died by the 18th day. When EBB(5 mg·kg-1·d-1) is jointly used with 5-FU (25 mg·mr-1d-1), 73%of mice with ascites H22 survived, much higher than 27%in the 5-FU treated group. EBB can enhance the anti-hepatoma ability of 5-Fu treatment. EBB mechanism against hepatoma: P53 expression in the EBB treated group is substantially higher than that in the control group. EBB increased the translation of P53. As a calmodulin antagonist, EBB decreases amount of the CaM in hepatoma cells and blocked the hepatoma cell proliferation cycle at the G2M phase. Before the G0/G1 phase, a diploid peak and apoptic cells in the treated groups were observed. Conclusions The CaM antagonist, EBB, has a strong anti-hepatoma effect and enhances the effect of 5-FU, induces hepatoma cell to apotopsis, promotes the P53 protein expression and decreases the amount of CaM in the cytoplasm. All these results demonstrate that EBB is a new and potentially useful drug against hepatoma and should be researched further. Chin Med J 2002; 115(5): 759-762

Objective To investigate the role of a potential diabetes-related mitochondrial region, which includes two previously reported mutations, 3243A→G and 3316G →A, in Chinese patients with adult-onset type 2 diabetes. Methods A total of 277 patients and 241 normal subjects were recruited for the study. Mitochondrial nt 3116- 3353, which spans the 16S rRNA, tRNAleu(UUR) and the NADH dehydrogenase 1 gene, were detected using polymerase chain reaction (PCR), direct DNA sequencing, PGR-restriction fragment length polymorphism and allele-specific PCR. Variants were analyzed by two-tailed Fisher exact test. The function of the variants in 16S rRNA were predicted for minimal free energy secondary structures by RNA folding software mfold version 3. Results Four homoplasmic nucleotide substitutions were observed, 3200T→C, 3206C→T, 3290T→C and 3316G→A. Only the 3200T→C mutation is present in the diabetic population and absent in the control population. No statistically significant associations were found between the other three variants and type 2 diabetes. The 3200T→C and 3206C →T nucleotide substitutions located in 16S rRNA are novel variants. The 3200T→C caused a great alteration in the minimal free energy secondary structure model while the 3206C→T altered normal 16S rRNA structure little. Conclusions The results suggest that the 3200T→C mutation is linked to the development of type 2 diabetes, but that the other observed mutations are neutral. In contrast to the Japanese studies, the 3316G→A does not appear to be related to type 2 diabetes. Chin Med J 2002; 115(5): 753-758

Objective To study the features of vascular smooth muscle cell (VSMC) proliferation induced by endothelin-1 (ET-1). Methods VSMCs of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats were cultured and treated with ET-1. Basic fibroblast growth factor (bFGF) gene expression was measured using both Northern blot and an enzyme-linked immunoassay. Results ET-1 resulted in an increase in bFGF transcripts at 8 - 24 h; bFGF levels were significantly higher in VSMCs treated with ET-1 than in those not treated. However, VSMCs growth responses in SHR and WKY were different. Smooth muscle cells of SHR were hyper-responsive to ET-1. Maximal bFGF mRNA levels were elevated 3.5-fold at 4 h of stimulation in WKY and 8-fold at 8h in SHR4. Moreover, the proliferation of VSMCs induced by ET-1 was inhibited by antisense phosphorothioate oligodeoxynucleotides(10 μmol/L AS-bFGF) but not sense bFGF oligomers at the same concentrations, being reduced by 80%in SHR and 40%in WKY vs control, respectively. Furthermore, the effect of AS-bFGF oligomers on SHR SMC proliferation is significantly greater than on WKY SMC proliferation. Conclusion ET-1 may be required for exaggerated vascular growth responses in SHR and bFGF may be involved. Chin Med J 2002; 115(5): 750-752

Objective To clone the full length of renal cell carcinoma (RCC) related novel gene GYLZ-RCC18 and study its function. Methods SMART RACE technology was used to clone the full length of GYLZ-RCC18. RT-PCR was used to detect its expression in renal cell carcinoma tissue at different stages and grades. We transfected the antisense oligonucleotide of GYLZ-RCC18 to renal cell carcinoma cell line, GRC-1, and analyzed proliferation activity, growth rate, apoptosis, and mortality changes. Results The full length of GYLZ-RCC18 (GenBank accession number: BE825133) cDNA was about 3.5 kb. GYLZ-RCC18 had a higher expression in higher grades and stages of renal cell carcinoma than in lower ones. The expression of GYLZ-RCC18 in renal cell carcinoma was much higher than in normal kidney. After the transfection of GYLZ-RCC18 antisense oligonucleotide, the mortality of GRC-1 increased significantly, while proliferative activity and growth rate were substantially inhibited at the same time. The antisense oligonucleotide induced apoptosis of GRC-1 through the entire observation time. Conclusion GYLZ-RCC18 is an important novel gene related to renal cell carcinoma. Overexpression of this gene results in higher growth and proliferative activity and has an antiapoptosis effect on renal cell carcinoma cells. Transfection of the antisense oligonucleotide may inhibit the generation and development of renal cell carcinoma. Chin Med J 2002; 115(5): 746-749

Objectives To detect the level of matrix metalloproteinase-2 (MMP-2) mRNA and the tissue inhibitor of metalloproteinase-2 (TIMP-2) mRNA in bladder transitional cell carcinoma (BTCC), and to estimate the prognosis for bladder tumor based on the quality and quantity of MMP-2 and TIMP-2 mRNA. Methods Thirty-five samples of human BTCC and 15 normal fresh bladder tissues were studied by RTPCR analysis followed by computer-assisted image analysis. Results The level of the MMP-2 mRNA in BTCC was significantly increased compared with that in normal bladder epithelium. The positive rates of MMP-2 and TIMP-2 mRNA were 71.4%and 65.7%in BTCC,and 66.7%and 60.0%in the normal bladder wall. The expression intensity of the MMP-2 mRNA by image analysis tended to increase with tumor grading and staging, which showed statistical significance. Similarly,the MMP-2 to TIMP-2 ratio also showed statistically significant difference between normal bladder tissue and bladder carcinoma (P <0.01) . Conclusions A high level of the MMP-2 mRNA exists in BTCC, which may function to damage collagenⅣ inside the basement membrane and the extracellular basement of the bladder. The level of the MMP-2 mRNA is proportional to BTCC grading and staging, which may have prognostic value. The MMP-2 to TIMP-2 ratio may play a more significant role in the determination of the aggressiveness and prognosis of bladder tumor. Chin Med J 2002; 115(5): 743-745

Objective To investigate gene expression of three nitric oxide synthase isozymes in injured spinal cord tissue. Methods Thirty-six adult SD rats were randomly divided into six groups: a normal group and five injury groups, with six per each group. Animals in the injury groups were sacrificed at 2, 6, 12, 24, 48 h after injury. A compression injury model on the spinal cord was made according to Nystrom B et al and gene expression of the three NOS isozymes were examined by reverse transcription polymerase chain reaction(RT-PCR). Results Gene expression of nNOS and eNOS were detectable in the normal group and were up-regulated quickly after injury, reaching a maximum at 6 h: (0.633±0.012) and (1.236±0.207). Gene expression of iNOS was detectable only in the injury groups and it was gradually up-regulated after injury, reaching a maximum at 24 h: (1 . 043 ± 0.049). Conclusion Injury to the spinal cord leads to early up-regulation of cNOS and late up-regulation of iNOS. Different NOS isozymes may play different roles in secondary spinal cord injury. Chin Med J 2002; 115(5): 740-742

Objective To study the clinical value of radical resection of gastric carcinoma with pancreas and spleen preservation (PSP) and functional cleaning of lymph nodes (LNs) of the spleen hillus and along the splenic artery. Methods Pancreas and spleen involvement was retrospectively reviewed among 439 cases of resectable carcinoma of the gastric cardia, gastric corpus and total stomach. During gastric surgery, 2 ml of methylene blue was injected into the subserosal space of the gastric cardia or corpus to observe the spread of lymphatic flow in 54 cases of gastric carcinoma. The metastatic rate of LNs in splenic hillus and along the trunk of the splenic artery (No10, No11), postoperative complications and survival rates were investigated in 63 gastric carcinoma patients that had received gastrectomy with pancreas and spleen preservation(PSP) . These were compared with the pancreas preservation (PP) group and pancreas and spleen combined resection (PSR) group. Results Among these 439 cases, only 25 cases were observed with direct invasion to the pancreas(5.7%), and 10 cases with direct invasion to the spleen (2.3%). After pathological examination of the pancreatic body and tail, we found 22 cases with pancreas and spleen combined resection, 4 cases(18.2%, 4/22) with direct invasion of the capsule and 2 with invasion to the superficial parenchyma(9.1%, 2/22), without metastasis to the lymph nodes within the pancreas and spleen. The metastatic rate of No10, No11 lymph nodes were 17.5%(11/63) and 19.1%(12/63) in the PSP group, 20.8%(45/216) and 25%(54/216) in the PP group, and 20%(6/30) and 23.3%(7/30) in the PSR group. There were no statistically significant differences (P> 0.05) . Injection of methylene blue into the subserosal space of the stomach did not diffuse into the spleen or pancreatic parenchyma. Postoperative complications diabetes and mortality in PSP (0%, 0%, 0%) were lower than in PP (4.2%, 0.9%. 0.9%) or PSR (40%,10%,3.3%). The 5-year survival rate (5-YSR) and 10-YSR in PSP (57.5%, 52.0%) were higher than in PSR (37 . 5%, 30 . 0%) . Those patients with stage Ⅱ and Ⅲa treated by PSP,improved markedly. Conclusions The surgical procedure of pancreas and spleen preservation for gastric cancer is a safe and organ function protected method. Postoperative complications were lower and survival rates were higher,the radicality was not reduced. These results indicate that PSP is preferred in patients with gastric carcinoma of stage Ⅱ or Ⅲa. Chin Med J 2002; 115(5): 736-739

Objective To observe the effects of recombinant human growth hormone (rhGH) on graft structure and recipient protein metabolism in rat small bowel transplantation (SBT) and total parenteral nutrition (TPN)models. Methods Twenty recipients of rat allogeneic heterotopic small bowel transplants (SD → Wistar) were divided into two groups (GH group and control group) Both groups were supported by standard TPN. Acute rejection was suppressed with CsA 10 mg·kg-1 ·d-1 intramuscularly. All rats in the experimental group received subcutaneous rhGH 1 U • kg-1 • d-1 after transplantation. Morphological mucosal indices of transplanted gut and metabolic parameters such as body weight, nitrogen balance, urinary 3-methyl histidine excretion and serum albumin of the recipients were compared between two groups. Results The application of rhGH promoted graft recovery significantly compared with standard TPN support alone. On postoperative day 14, all morphological indexes of transplanted gut recovered to the preoperative state. Protein metabolism in the recipient was also significantly improved. rhGH decreased the catabolism of protein, accelerated regaining of positive nitrogen balance and corrected hypoalbuminemia. Conclusion GH is an effective metabolic intervention in SBT. It may promote the structural repair of the graft and correct the metabolic disturbance. It is useful in improving the outcome of clinical SBT. Chin Med J 2002; 115(5): 732-735

Objective To define the mechanism of acute hepatitis in non-human primates after liver directed gene therapy . Methods Differences in immune response exhibited by 8 rhesus monkeys receiving adenovirus (Ad) or lipofectamine-mediated gene transfer by various routes, the time course, and the nature of the specific immune responses to both adenoviral vectors and transgene products were studied using HE staining(H&E) and immunohistochemical staining. Results The monkeys developed mild to moderate acute hepatitis 1 to 3 weeks after intravenous or intrabiliary injection of first generation replication-defective adenoviruses carrying the Escherichia coli lacZ gene. This was accompanied by adenovirus-mediated T-cell proliferation and neutralizing antibodies to the adenovirus. Increased numbers of CD3+, CD4+ and CD8+T-lymphocytes were detected in the diseased livers, while B-lymphocytes were absent. Hepatocytes demonstrated increased expression of β2-microglobulins (β2-MG) and HLA-DR antigens in the plasma membranes. The development of acute hepatitis and the accompanying immune abnormalities were delayed in immunosuppressed monkeys until after the discontinuation of immunosuppressive therapy. The monkeys infused with Ad. CMVluc showed more significant and longer durations of hepatitis than the monkeys infused with adenoviruses carrying the lacZ gene. Lipofectamine-mediated gene transfer was inefficient. There was neither lacZ expression nor significant immune response in the liver of monkeys infused with lipofectamine via the portal vein or the common bile duct. Conclusion Immune response to the hepatocytes in liver directed gene therapy is MHC class I restricted and T-cell mediated. Both adenoviral vectors and foreign genes are related to the liver damage. Mild to moderate hepatic inflammation seen with the E-1 deleted vector is reversible. Immunosuppression regimens may prolong transgene expression and delay the development of acute adenoviral hepatitis. Chin Med J 2002; 115(5): 726-731