Volume 115  ·   Issue 04  ·  2002

Objective To establish a profile of the causes of apparently unexplained SS in genetic referral center and evaluate the current referral system. Methods This was a retrospective database survey on patients who were referred our clinical genetic service from 1988 - 1998 primarily because of SS. We retrieved the study population from our computer database using "short stature" as a search handle and then studied the demographic, clinical and laboratory data from their medical records. Results Three hundred and fifty-three subjects were referred for genetic evaluation of SS in 1988- 1998. The mean age of referred subjects was 11.5 years and the female to male ratio was 7.6. All referrals had undergone cytogenetic studies to exclude chromosomal abnormalities, 19%of girls with apparently unexplained short stature had Turner syndrome; at least 47.9%of the study population were normal variants and 25%of the referrals had inadequate information for classification. Conclusions Genetic investigation is essential in the management of patients with SS, especially for girls suspected of having Turner syndrome, in which growth hormone treatment has shown to improve final height. We also highlight the inherited causes of short stature, which were often misdiagnosed as benign familial short stature, and discussed the drawbacks of the current referral system. Chin Med J 2002; 115(4): 607-611

Objective To explore the association of inherent cellular reactive oxygen species (ROS) levels with susceptibility of the tumor cells to apoptosis induction by arsenic trioxide (As2O3). Methods Low concentration (2 μmol/L) of As2O3 was administered to two cultured leukemic cell lines,NB4 and U937, and two esophageal carcinoma cell lines, EC1 .71 (also named EC/CUHK1) and EC1867,to confirm the difference in apoptosis susceptibility of NB4 versus U937 and of EC1 .71 versus EC1867. Dihydrogenrhodamine 123 (DHR123), used as a ROS capture agent, was incubated with cells in the absence of As2O3. Fluorescence intensity of rhodamine 123, the product of cellular oxidation of DHR123,was detected by flow cytometry and ROS was measured. Results Low concentration of AS2O3 induced apoptosis was more likely to occur in NB4 and EC1 .71 cells than in U937 and EC1867 cells, or NB4 was more sensitive than U937, and EC1 .71 more sensitive than EC1867 to AS2O3. The inherent cellular ROS level is higher in NB4 than in U937, and also higher in EC1 .71 than in EC1867. Conclusions The difference in cellular ROS level is positively associated with cellular susceptibility to apoptotis induction by AS2O3. The inherent ROS level might be important in defining apoptotic susceptibility to AS2O3 . Chin Med J 2002; 115(4): 603-606

Objective To study the effect of hepatitis C virus nonstructural protein NS3 (HCV NS3) on telomerase activity and carcinogenesis. Methods Streptavidin-peroxidase (SP) conjugated method was used to detect the expression of HCV NS3 protein in NIH3T3 cells transfected with plasmid pRcHCNS3-5' and pRcHCNS3-3'. Telomerase activity was detected by an in situ telomerase activity labeling method, telomeric repeat amplification protocol polymerase chain reaction (TRAP-PCR) and telomerase PCR enzyme linked immunosorbent assay(ELISA) technology in the transfected and non-transfected NIH3T3 cells. Results HCV NS3 protein was expressed in the NIH3T3 cells transfected with plasmid pRcHCNS3-5 ' expressing HCV NS3 C-terminal deleted protein or with plasmid pRcHCNS3-3 ' expressing HCV NS3 Nterminal deleted protein. The positive signal of HCV NS3 protein was localized in the cytoplasm of NIH3T3 cells, and the signal intensity of the former was stronger. Telomerase activity in NIH3T3 cells transfected with plasmid pRcHCNS3-5' was stronger than that in NIH3T3 cells transfected with plasmid pRcHCNS3-3'(P <0.01), whereas telomerase activity in NIH3T3 cells transfected with plasmid pRcCMV or untreated NIH3T3 cells was weaker than that in NIH3T3 cells transfected with plasmid pRcHCNS3-3' (P <0.05). The expression level of HCV NS3 protein was significantly correlated with the strength of telomerase activity(P <0.05). The results obtained by in situ telomerase activity labeling corresponded to the results by telomerase PCR ELISA technology. Conclusions HCV NS3 protein may activate telomerase through endogenous mechanism to induce host cell transformation. The effect of HCV NS3 C-terminal deleted protein on telomerase activity in the host cell may be stronger than that of HCV NS3 N-terminal deleted protein. In situ telomerase activity labeling was a reliable technology for studying pathological morphology and telomerase activity in tissues and cells. Chin Med J 2002; 115(4): 597-602

Objective To evaluate the protective effects and mechanism of action of oxymatrine (OM) on the experimental fulminant hepatitis (FH) and early hepatocyte apoptosis in murine liver tissue. Methods Fulminant hepatitis mice were induced by injecting lipopolysaccharide (LPS) intraperitoneally(ip) in galactosamine (GaIN) sensitized mice. Two separate experiments were designed, including saline control group, fulminant hepatitis group and oxymatrine pretreated group (50 mg/kg, intraperitoneally,bid × 3 days). The levels of serum tumor necrosis factor alpha (TNFa) in mice from two experiments were determined at 5-hour and 7.5-hour after injecting galactosamine/lipopolysaccharide. Mouse liver samples at 5-hour time point were obtained for in situ end labeling (ISEL) staining and ultrastructural observation of apoptotic cells under transmission electron microscope (TEM). Liver samples at 7.5-hour time point were taken for hematoxylin-eosin (HE) staining and immunohistochemical staining of Fas and its ligand (FasL). Results As compared with the fulminant hepatitis group, the levels of serum tumor necrosis factor alpha in mice from the OM pretreated group at 5-hour and 7.5-hour time point were all significantly decreased (P < 0.05 and P <0.01 respectively) . Hepatocyte apoptosis in mice at 5-hour time point was significantly inhibited (P <0.01). Both the degree of liver injury and the degree of Fas and Fas ligand expression in the OM pretreated group were reduced remarkably (P <0.01 and 0.05 respectively) when compared with the saline control group. Conclusions Oxymatrine protects mice from fulminant hepatitis induced by GalN/LPS and may block hepatocyte apoptosis and subsequent necrosis through downregulating the production of serum tumor necrosis factor alpha and the expression of Fas and Fas ligand in liver tissue. Chin Med J 2002; 115(4): 593-596

Objective To investigate the inherent characteristics of Vibrio cholerae (V. cholerae) and other vibrios and their relationship. Methods Polymerase chain reaction (PCR), DNA sequence analysis, randomly amplified polymorphic DNA (RAPD) analysis and average linkage cluster analysis were used to study 3 isolates of V. cholerae strains 0139, three isolates 01 biotype El Tor, four isolates 01 biotype classical and 3 other vibrios. Results V. cholerae 0139 contained the genomic sequences of ctx A2-B as well as V. cholerae 01. V. cholerae and others vibrios were divided into 4 groups by fingerprint patterns of RAPD, that is ①V. cholerae 0139 and V. cholerae 01 El Tor; ② V. cholerae 01 classical; ③ V. paraheamolyticus and V. vulnificus and ④ V. flluvialis. V. cholerae 0139 DNA fingerprint of RAPD was consistent with the El Tor biotype: average linkage cluster distance was 0, and slightly different from the classical biotype, with a distance of 2.07. It was much more different from vibrio paraheamolyticus and others, with a distance of 6.76-8.54. Conclusion V. cholerae and other vibrios are polymorphic in inherent characteristics. The inherent characteristics of V. cholerae 0139 are the same as El Tor biotype. 0139 may have evolved from the El Tor biotype. The inherent characteristics of vibrio paraheamolyticus are the same as vibrio vulnificus. Chin Med J 2002; 115(4): 589-592

Objective To study the inhibition effect of oxLDL on the uptake and clearance of intra-cellular 3H-cholesterol in v-SMC from the human-apoA1 transgenic mice (C57BL/6) and the changes in humanapo A1 mRNA expression in v-SMC from human apoAl transgenic mice after oxLDL stimulation. Methods v-SMC originally isolated from human apoAl transgenic mice connected with a recombined mouse metallothionein-I (MT-I) promoter was used, and the effect of oxLDL on the uptake and clearance of intracellular 3H-cholesterol was studied in v-SMC of the transgenic and control mice respectively, the study of h-apoA I mRNA expression from v-SMC of the transgenic mice were done by RT-PCR and Northern blot. Results oxLDL (30 μg/ml) strongly promoted the SMC proliferation. No difference was found in 3 H-cholesterol uptake between nSMC and trSMC, and the uptake rates of both kinds of SMC rose 100%after oxLDL stimulation. The efflux rates of 3H-cholesterol in trSMC were much higher than those of nSMC(40%-50%). After oxLDL stimulation, the clearance rates fell by 28%and 10%, respectively, for nSMC and trSMC. The result of RT-PCR and Northern blot showed that h-apoA1 gene expression was markedly increased by the stimulation of oxLDL. Conclusion Expression of the h-apoA1 gene in C57BL/6 mice enables them to reduce the accumulation of cholesterol in v-SMC. The trSMC can alleviate the harmful effect of oxLDL due to the increase of h-apoA1 expression. Chin Med J 2002; 115(4): 584-588

Objective To probe the significance of specific IgG4 in sera of patients with cerebral cysticercosis for diagnosis and therapeutic evaluation. Methods Specific IgG4 in sera of patients with cerebral cysticercosis was assessed using colloidal gold-labeled mouse-anti-human IgG4 McAb as probe. The results were compared with the CT image manifestation. Results The specific IgG4 positive rate in sera of patients with cerebral cysticercosis was 97.8%, whereas sera from patients with other kinds of parasitosis or central nerve system disease and the control group were all negative, except for a weak cross-reaction of sera from patients with hepatic echinococoosis. The determination of specific IgG4 in sera of patients with cerebral cysticercosis during different times of treatment showed that along with an increase in treatment time and improvement of clinical symptoms, specific IgG4 level gradually decreased. The positive rate and intensity of specific IgG4 in sera from patients with cerebral cysticercosis were consistent with the number of cysticercus parasites in the brain and pathologic changes, such as survival, disintegration, death and calcification. Survival of cysticercus in the brain was objectively evaluated using this technique. Conclusions The determination of specific IgG4 in sera is a practical method for diagnosis and therapeutic evaluation of cerebral cysticercosis. Chin Med J 2002; 115(4): 580-583

Objective To investigate the protective immunity induced by the anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum in mice . Methods An orthogonal table L16 (4 x 212) was selected as the experimental design. Eight-week-old Kunming outbred mice (male and female) were randomly divided into 16 experimental groups and 2 control groups. Control groups were injected with SP2/0 ascites intraperitoneally. Mice from each group were infected with 100 ±2 cercariae of Schistosoma japonicum in the abdominal skin and were sacrificed on the thirtieth day postchallenge. Adult worms were recovered and counted by perfusion of the left ventricle-portal vein. The SP2/0 ascites injected mice were used as controls and the percentage of protection was calculated. Results Active immunization of mice with NP30 could produce protection levels ranging from 22.36%to 50.46%depending on the different immunity protocols. The best immunization protocol was established from the results. Conclusions Active immunization with NP30 can induce a degree of protection to infection with Schistosoma japonicum cercariae and NP30 is a potential vaccine candidate against Schistosoma japonicum . Chin Med J 2002; 115(4): 576-579

Objective To investigate the loss of heterozygosity (LOH) on chromosomal arms 13q and 14q in nasopharyngeal carcinoma (NPC) using 21 microsatellite polymorphic markers and to study whether there is a correlation between LOH and clinicopathologic parameters and/or Epstein-Barr virus (EBV) infection in NPC. Methods Sixty cases of NPC were studied using polymerase chain reaction based microsatellite analysis with genescan and genotyping techniques. Results LOH was detected on 13q in 78%of NPC tumors, high frequency LOH loci (more than 30%)clustered to 13q12.3-q14.3 and 13q32. On chromosome 14q, LOH was detected in 80%of NPC tumors;high frequency LOH loci clustered to 14q11-q13, 14q21-q24 and 14q32. High frequency LOH at 13q31-q32 correlated with a lower level of EBV infection; LOH on chromosome 14q was closely associated with poor differentiation of NPC tumor cells. Conclusion Our results suggest that in NPC, LOH on chromosome 13q and 14q are common genetic events, and putative tumor suppressor genes (TSG) residing in these regions may be involved in tumorigenesis. Chin Med J 2002; 115(4): 571-575

Objective To investigate the effect of active immunotherapy with anti-idiotypic vaccine in patients with nasopharyngeal carcinoma (NPC). Methods Anti-idiotypic antibodies (2H4/5D3) bearing the internal image of the NPC antigen were used in active immunotherapy in NPC patients receiving radiotherapy. Antibodies and cytokine levels in patient sera were determined using ELISA before and after active immunotherapy. IL-2 mRNA expression in the peripheral blood mononuclear cells (PBMC) was measured by in situ hybridization. Results Nineteen patients with NPC at stage Ⅳ were treated with alum-precipitated 2H4 or 5D3. Neither hypersensitivity nor adverse side effects were observed. The levels of anti-anti-idiotypic antibodies (Ab3)and anti-NPC antibodies (Ab1’) were increased. Human anti-mouse antibodies (HAMA) were seen in 19 patients of the experimental group; the levels of Ab1' did not increse in the control group. Serum IL-2,IFN-γ and TNF-a levels were increased in most patients in the experimental group, while no differences were observed in Ab1' and cytokine levels between pre- and post-therapy in the control group. In addition,IL-2 mRNA expression in PBMCs from NPC patients was closely related to serum IL-2 (r = +0.8829)levels by in situ hybridization. Conclusions Anti-idiotype vaccine is safe for clinical active immunotherapy. Anti-idiotypic vaccine might be able to enhance humoral and/or cellular immunity in NPC patients receiving radiotherapy. Chin Med J 2002; 115(4): 567-570

Objective To study the influence of intranasal medication on the structure, pathology and reversibility of the nasal mucosa to provide a basis for the feasibility of intranasal route of drug administration. Methods Nasal drops of gentamicin were placed in the nasal cavity of rabbits for 3, 5, 7, 14 and 28 days. After that, the drops were stopped and drugs protecting the nasomucosa were used for one and three weeks. After being sacrificed over time, the nasomucosa of the rabbit was observed under optical and electron microscopes. Results Damage to the nasal mucosa appeared to different extents with prolonged use of nasal drops. Within 3-7 days of applying the drug, damages to the nasal mucosa gradually appeared, and after two and four weeks, were most serious. After stopping the drug, the nasal mucosa was gradually restored. Conclusion Damages of drugs to the nasal mucosa could be restored. The intranasal route of drug administration would be feasible and clinically applicable. Chin Med J 2002; 115(4): 617-619

Objective To investigate the effect of polysaccharide sulfate 916 (PS916) on the production of nitric oxide(NO) in ECV304 cells induced by tumor necrosis factor-a (TNFcx), interleukin-iβ (IL-1β) and H2O2 in vitro. Methods Production of NO in ECV304 cells was measured by the Griess method and the proliferation of cells was tested by the MTT method. The activity of NO synthase was detected spectrophotometrically. Results Production of NO in ECV304 cells decreased after treatment with 40 ng/ml IL-1β and 40 ng/ml TNFα, but increased in the presence of H2O2 0.1 mmol/L. PS916 significantly enhanced NO production in ECV304 cells in a dose-dependent manner in the TNFα and IL-1β treated groups and decreased it in the H2O2 treated group. Proliferation of ECV304 cells was inhibited by TNFβ and H2O2 and no effect was found in the IL-1β treated group. PS916 increased the proliferation of cells treated with TNFα and H2O2 dose-dependently. In vitro, PS916 has no effect on the activity of NO synthase. Conclusion PS916 has a protective effect on ECV304 cells exposed to IL-1β, TNFα and H2O2. Chin Med J 2002; 115(4): 614-616

Objective To investigate the feasibility and features of 3 catheter-mediated approaches of gene transfer into heart, including direct myocardial injection (DMI), coronary artery perfusion (CAP), and intrapericardial cavity injection (ICI). Methods Fifteen dogs were used, and 0.3 ml (1 x 109 pfu) of an adenovirus (Adex1SR LacZ) was injected into the heart by 3 methods. The dogs were killed 5 days following injection, and gene expressions in heart and liver were evaluated by histochemical analysis. Results The results showed that ① the CAP method was relatively less damaging and induced sparse LacZ expression in the myocardium, and the gene expression was also found in both vessels within the myocardium and liver; ② gene transfer by DMI resulted in intense LacZ expression around the injection accompanied by a local inflammatory response; ③ LacZ expression elicited by ICI was detected in either the inner surface of the parietal pericardium or epicardial surface of the heart, and also in the myocardium underlying the visceral pericardium. Conclusion Three catheter-mediated methods of gene transfer into the heart may be used and a reasonable approach should be chosen according to purpose. Chin Med J 2002; 115(4): 612-613