Child, Preschool ; Female ; Humans ; Sarcoma ; diagnosis ; surgery ; Stomach Neoplasms ; diagnosis ; surgery ; Stromal Cells ; pathology

OBJECTIVETelomerase, a complex of ribose and nucleoprotein, is a specific marker of tumor, which expresses in 98% infinite cell lines and 90% malignant tumor organizations and whose function is to maintain the length of telomere. Human telomerase reverse-transcript protein subunit (hTERT) is the key element and rate-limiting factor of telomerase activity. Our study was to investigate the effects of antisense hTERT gene on biological characteristics of hepatoblastoma cell line in vitro.

METHODSThe sense and antisense hTERT eukaryotic expression vectors that we had constructed before were transfected into hepatoblastoma cell line HepG2 by using the SuperFect transfection reagent (Qiagen) according to the manufacturer's instructions, then the HepG2-s and HepG2-as of G418-resistant colonies were obtained with G418 and identified for the presence of hTERT insert by PCR with T7 and pcDNA3.1/BGH reverse primers. After that, we have detected the endogenous hTERT mRNA expression and telomerase activity by quantitative real-time RT-PCR and TRAP-silver staining assay in cells from each group. Meanwhile, MTT cellular proliferation assay, soft agar colony formation assay and flow cytometry were employed to analyze if the proliferation capacity of liver cancer cells was affected in vitro and the tumor cells could be induced to apoptosis by antisense hTERT.

RESULTSAntisense hTERT significantly down-regulated the endogenous hTERT mRNA expression (15.35 +/- 1.72/HepG2-as, 43.8 +/- 2.89/HepG2-s, 45.2 +/- 3.46/HepG2) (n = 10, t = 7.61, P < 0.01) and telomerase activity in HepG2, compared to blank control and sense hTERT. After 20 passages of three group cells, a 7-day cell growth curve and the numbers (size) of soft agar colony formation showed the proliferation and the anchorage-independent growth in HepG2-as were significantly suppressed (50.6 +/- 4.8/HepG2-as, 113.52 +/- 8.15/HepG2-s, 119.12 +/- 10.82/HepG2) (n = 10, t = 4.54, P < 0.01 and n = 10, t = 3.96, P < 0.01), compared to HepG2 and HepG2-s. However there was a significant increase in apoptosis percentage of HepG2-as by flow cytometry (n = 10, t = 9.24, P < 0.01 and n = 10, t = 8.37, P < 0.01), compared to control group.

CONCLUSIONSAntisense hTERT could significantly suppress the hepatoblastoma cell growth and reverse its malignant phenotypes in vitro and cause the increase in apoptosis percentage of HepG2, thus it might be applied in malignant tumor gene therapy through the telomerase-targeted molecular mechanism.

Cell Division ; genetics ; Cell Line, Tumor ; DNA-Binding Proteins ; Hepatoblastoma ; genetics ; pathology ; Humans ; RNA, Antisense ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Telomerase ; genetics

OBJECTIVETo investigate the effect of the sera of rabbits fed with Tongxinluo on the expression and secretion of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in U937 monocyte-derived macrophages.

METHODSAtherosclerosis was induced in rabbits by high-cholesterol feeding, and the serum was obtained from the rabbits after administration of the aqueous solution of Tongxinluo or simvastatin by gavage. U937 monocyte-derived macrophages were incubated with the sera at different concentrations for 24 hours, and the changes in MMP-9 and TIMP-1 gene expression and secretion were detected by RT-PCR and enzyme-linked immunosorbent assay, respectively.

RESULTSThe serum of rabbits fed with Tongxinluo concentration-dependently inhibited the expression and secretion of MMP-9 in U937 macrophages, but did not affect TIMP-1 expression or secretion.

CONCLUSIONTongxinluo may stabilize the atherosclerotic plaques by inhibiting the expression and secretion of MMP-9.

Animals ; Atherosclerosis ; blood ; etiology ; Cholesterol, Dietary ; administration & dosage ; Drugs, Chinese Herbal ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Macrophages ; drug effects ; metabolism ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Serum ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; U937 Cells

OBJECTIVEKawasaki disease (KD) is an acute febrile vasculitic syndrome of unknown etiology that preferentially affects coronary artery. It has been suggested that proinflammatory cytokines like tumor necrosis factor alpha (TNF-alpha) and interleukin-10 (IL-10) are key players during acute KD. Recently, the polymorphisms relative to major transcriptional start site of TNF-alpha and IL-10 gene were shown to influence the level of TNF-alpha and IL-10 production in vitro. This study was aimed to investigate the genetic association of TNF-alpha and IL-10 promoter polymorphisms in juvenile patients of Han nationality with KD, and to investigate the possible associations with clinical manifestations of the disease.

METHODSFour polymorphism sites of TNF-alpha and IL-10 gene promoter regions from 96 children with KD were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). One hundred and sixty age-matched normal children of the Han nationality were used as control. All patients accepted Doppler echocardiography examination in order to differentiate coronary artery lesions.

RESULTSThere was significant difference in allele frequencies of -308 (A/G) site of the TNF-alpha gene between children of the Han nationality and those of Japanese and Caucasian in America. There were significant differences in the allele frequencies of -1082 (G/A), -819 (C/T) and -592 (A/C) of IL-10 gene between children of the Han nationality and their British Counterparts (P < 0.01). There was no significant difference in allele frequencies of -308 (A/G) site of TNF-alpha gene between children with KD and normal controls. There was no significant difference in the haplotypes and the allele frequencies of the above three sites of IL-10 between the two groups. However, when clinical features were examined, the genotype frequency of TNF-alpha-308A was significantly higher in IVIG-resistant KD patients than that of TNF-alpha-308G genotype (67% vs 5%, chi(c)(2) = 90.48, P < 0.01). The genotype of TNF-alpha-308A was closely associated with IVIG-resistant KD (P < 0.01, relative risk 42.25, 95% confidence interval 15.81-112.88). The haplotype frequency of IL-10 -1082A/-819T/-592A was also higher in patients with coronary artery lesion (CAL) caused by KD than those of Non-ATA haplotype (52% vs 20%, chi(2) = 18.36, P < 0.01). The haplotypes of IL-10 -1082A/-819T/-592A was significantly associated with CAL caused by KD (P < 0.01, relative risk 4.26, 95% confidence interval 2.20-8.25).

CONCLUSIONThe genotype of TNF-alpha-308A is one of the important factors that probably influence the therapeutic effect of KD. The haplotypes (-1082/-819/-592) of IL-10 gene promoter might be related to the pathogenesis of coronary artery complication of KD and -1082A/-819T/-592A haplotypes might be regarded as a genetic marker of risk factor for coronary artery lesion in KD.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Interleukin-10 ; genetics ; Male ; Mucocutaneous Lymph Node Syndrome ; genetics ; pathology ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Promoter Regions, Genetic ; genetics ; Tumor Necrosis Factor-alpha ; genetics

BACKGROUNDCockroaches are an important indoor allergen source causing allergic rhinitis and asthma. The aim of this study was to investigate the cockroach prevalence in mainland of China and the cross-reactivity of IgE between cockroach and house dust mite allergen in Chinese patients.

METHODSThe cockroach sensitization pattern was based on a skin prick test (SPT) obtained from a national multicenter prevalence study, in which 6304 patients from 25 allergy centers across China participated. Factors, including different regions of China, age, gender and the correlations between the American and German cockroaches and house dust mite Der p were investigated. Eighteen out of 1236 clinical sera from south China were selected to perform the cross-inhibition assay between house dust mites and cockroaches.

RESULTSTotally 25.7% of patients were SPT positive to the American cockroach (Periplaneta Americana, Per a) and 18.7% SPT positive to the German cockroach (Blattella germanica, Bla g). The prevalence of positive cockroach SPT was higher in southern than in northern China, higher in adults than in children, and higher in males than in females. Patients had relatively low levels of cockroach SPT reactions, mainly class 1 or 2. Of the SPT positive cockroach patients, 88% were also SPT positive to house dust mite (Dermatophagoides pteronyssinus, Der p). An IgE cross-inhibition study confirmed that Der p sensitization could cause false positive SPT reactions against cockroach.

CONCLUSIONSA relatively high prevalence of cockroach sensitivity was found in mainland of China. However, a cross-inhibition study showed that only a small number of patients appear to have Bla g and/or Per a as primary sensitizing source. The importance of cockroaches as a risk factor for sensitization and triggers of allergic symptoms in mainland of China needs to be further investigated.

Adolescent ; Adult ; Aged ; Allergens ; immunology ; Animals ; Antigens, Dermatophagoides ; immunology ; Asthma ; etiology ; Child ; Child, Preschool ; China ; epidemiology ; Cockroaches ; immunology ; Cross Reactions ; Female ; Humans ; Hypersensitivity ; epidemiology ; Immunoglobulin E ; immunology ; Male ; Middle Aged ; Prevalence ; Rhinitis, Allergic, Perennial ; etiology ; Rhinitis, Allergic, Seasonal ; etiology

Objective To compare the bio-medical parameters in SIV infected Chinese rhesus monkeys with diverse disease progression,by which the pathogenesis of simian AIDS were to be investigated.Methods Sixteen Chinese rhesus monkeys were inoculated intravenously with SIVmac239 and followed-up for 18 months.Based on their progression patterns and plasma viral loads,animals were divided into 3 groups,including 1 rapid progressor( RP),13 normal progressors(NP),and 2 elite controllor(EC).Their parameters of haematology,virology,immunology and pathology were examined and compared. Results Compared with other animals,RM449(RP) showed higher viral load,unresponsive humoral immunity,and higher level of auto-antibodies against lymph node,thymus,and spleen.Additionally,its effector memory CD4 count was lower,with the transformation progress being blocked-like from naive/central memory subsets to effector memory subset,as the flow-cytometry assay showed.Notable decrease in its peripheral B cell was also observed,especially to the sub-population of tissue-like memory B cells and activated memory B cells.Pathological examination showed the depletion of lymphoid tissue,atrophy of spleen and loss of thymus.Moreover,most of these parameters of RM450 and RM453 (EC) changed opposite to that of RP.Conclusion The hallmarks of RM449 were higher viraemia and lower SIV specific IgG level,which may due to the disturbance of T cells and B cells development and differentiation.Moreover,destructions of organs of the immune system may contribute to the disturbance.Our study suggest that the change of micro-environments of thymus induced by SIV infection,which is necessary in T cell and B cell development and differentiation,may contribute at least partially to the AIDS pathogenesis.

Objective To observe the protective effects of a mixture for protecting liver and supplementing stomach (保肝益胃合剂) on rat acute liver damage induced by carbon tetrachloride (CCl_4). Methods The model of rat acute liver damage was established by injection of CCl_4 2 ml/kg into the abdominal cavity.The rat models were treated respectively by the mixture for protecting liver and supplementing stomach 30 g?kg~(-1)?d~(-1),the polyene phosphatide acid radical choline capsule [Yi Shanfu (易善复), 180 mg?kg~(-1)?d~(-1)],the glycyrrhizic acid diaminogen capsule [Gan Lixin (甘利欣),30 mg?kg~(-1)?d~(-1)] infused into the stomach.The activities of serum alanine transaminase (ALT) and aspartate transaminase (AST) were detected.In the mean time,the liver pathological changes were observed,the degree of liver cell necrosis was evaluated,and the rat mortality was noted in various groups of treatment.Results The values of ALT,AST and the score of liver cell necrosis in the group treated with the mixture for protecting liver and supplementing stomach [(1.168?1.066) kU/L,(1.845?2.212) kU/L,(0.56?0.53) score] were significantly lower than those in the model group [(4.982?3.502) kU/L,(7.030?3.616) kU/L, (1.38?0.92) scores],and all the differences being statistically significant (all P

OBJECTIVETo analyze the factors leading to prescheduled analgesic withdrawal in patients with postoperative epidural analgesia.

METHODSA retrospective study of 4876 patients with postoperative epidural analgesia was conducted and the effect of analgesia and incidence of prescheduled analgesic withdrawal were recorded. The factors precipitating the occurrences of analgesic withdrawal and complications were analyzed.

RESULTSEarly analgesic withdrawal occurred in 113 cases (2.3%), among which 74 (0.5%) were due to factors irrelevant to analgesic complications. Analgesia-related complications occurred in 578 patients, but only 39 (0.7%) of them needed discontinuation of the analgesics.

CONCLUSIONPrescheduled analgesic withdrawal is predominantly due to technical inadequacies rather than complications arising from the analgesics, and improvement of the operation skills for postoperative analgesia may reduce early analgesia discontinuation and enhance the patients' satisfaction.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Analgesia, Epidural ; statistics & numerical data ; Female ; Humans ; Male ; Middle Aged ; Postoperative Complications ; Postoperative Period ; Retrospective Studies ; Time Factors ; Young Adult

BACKGROUNDNucleus pulposus of intervertebral discs has proinflammatory characteristics that play a key role in neuropathic pain in lumbar herniated intervertebral disc. One of the most commonly used animal models (the traditional model) of non-compressive lumbar herniated intervertebral disc is created by L4-L5 hemilaminectomy and the application of autologous nucleus pulposus to cover the left L4 and L5 nerve roots in rats. However, such procedures have the disadvantages of excessive trauma and low success rate. We proposed a modified model of non-compressive lumbar herniated intervertebral disc in which only the left L5 dorsal root ganglion is exposed and transplanted with autologous nucleus pulposus following incision of epineurium. We aimed to compare the modified model with the traditional one with regard to trauma and success rate.

METHODSThirty Sprague-Dawley male rats were randomized into three groups: sham operation group (n = 6), traditional group (n = 12), and modified group (n = 12). The amount of blood loss and operative time for each group were analyzed. The paw withdrawal threshold of the left hind limb to mechanical stimuli and paw withdrawal latency to heat stimuli were examined from the day before surgery to day 35 after surgery.

RESULTSCompared with the traditional group, the modified group had shorter operative time, smaller amount of blood loss, and higher success rate (91.7% versus 58.3%, P < 0.05). There was no decrease in paw withdrawal latency in any group. The sham operation group had no decrease in postoperative paw withdrawal threshold, whereas the modified and traditional groups had significant reduction in paw withdrawal threshold after surgery (mechanical hyperalgesia).

CONCLUSIONSTransplantation of nucleus pulposus onto the L5 dorsal root ganglion following incision of epineurium in rats established an improved animal model of non-compressive lumbar herniated intervertebral disc with less trauma and more stable pain ethology.

Animals ; Disease Models, Animal ; Ganglia, Spinal ; pathology ; Intervertebral Disc Degeneration ; pathology ; Intervertebral Disc Displacement ; pathology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo summarize the experience of surgical resection of 103 hilar cholangiocarcinoma.

METHODSOne hundred and three consecutive cases of hilar cholangiocarcinoma who underwent surgical resection at our hospital over the past ten years were reviewed retrospectively. The clinical data and long-term outcome were analyzed.

RESULTSOut of 103 cases, 43 patients underwent radical resection (41.7%), and 60 patients underwent palliative resection. There were 34 patients developed postoperative complications and 8 patients died in hospital. For the radical resection group, the median survival time was 29.9 months and 1-year, 3-year, 5-year survival rate was 69.6%, 42.0%, 20.9%, respectively, which was significant greater than 34.1%, 10.2%, 0 of the palliative resection group (P < 0.05). Over the past five years, 42 cases underwent pre-operative drainage of bile and the rate of combined liver resection reached 53.8%. The tumor radical resection rate has increased to 45.7%, the median survival time have reached 24.7 months (P < 0.05).

CONCLUSIONSImprovement of pre-operative management, intraoperative pathology for resection margin, and combined liver resection may help in increasing the radical resection rate. Radical resection can improve postoperative survival, and produce a satisfactory outcome for patient with hepatic hilar cholangiocarcinoma.

Adult ; Aged ; Bile Duct Neoplasms ; mortality ; surgery ; Bile Ducts, Intrahepatic ; Cholangiocarcinoma ; mortality ; surgery ; Digestive System Surgical Procedures ; methods ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Survival Rate