Objective To evaluate the measurement of kappa and lambda immunoglobulin free light chains(FLC) in patient samples using a new enzyme-linked immunosorbent assay for early diagnosis of multiple myeloma.Methods An ELISA method for quantifying kappa and lambda free light chains were used to study serum and urine samples from patients with multiple myeloma, and the results were compared with those obtained using immunofixation electrophoresis and nephelometric immunoassay methods. Results The FLC-ELISA method had great successful rate in identifying the multiple myeloma in all 40 myeloma patients. In contrast,immunofixation electrophoresis and nephelometric immunoassay could only identify 57.5% and 85.5% of the multiple myeloma in all the myeloma patients,respectively. Furthermore, retrospective diagnosis of specimens obtained from patient indicated that the ELISA method could help early diagnosis of the disease by over two years. Conclusion The ELISA method for measuring free light chains is sensitive, accurate and reproducible. Therefore it is a useful tool for early diagnosis of multiple myeloma,monitoring the disease progression and evaluating treatment responses.

Objective To investigate the expression level of endogenetic nitric oxide(NO) in the reproduction process of human anterior cruciate ligament cells. Methods Anterior cruciate ligament cells were isolated and subcultured from the human anterior cruciate ligament. LPS was used to induce the anterior cruciate ligament cell to express the inducible nitric oxide synthase(iNOS ), and N-monomethyl -L-Arginine (L-NMMA) was used as the interdiction of nitric oxide. They were alone or together added in the culture medium of anterior cruciate ligament cells in different groups. The level of NO was indirectly measured in the medium of HACL. Results LPS promoted significantly anterior cruciate ligament cells to produce endogenetic nitric oxide, compared with the control group(P

AIM:To investigate the role of high glucose in primary hepatocytes of mice fed with a high fat di-et.METHODS:Male C57BL/6J mice were fed a high fat (45%of calories) diet ad libitum for 6 weeks to induce hepatic steatosis.Primary hepatocytes were isolated from the mouse liver by the 2 step collagenase perfusion method .The cells were incubated in low glucose ( 5 mmol/L ) , low glucose plus mannitol ( 30 mmol/L ) , or high glucose ( 35 mmol/L ) DMEM medium for 12 h.The cell viability , apoptosis , mitochondrial membrane potential , and caspase enzymatic activities were measured.Furthermore, proteins related to the stress-sensitive signaling pathway of regulating high glucose-induced apoptosis in primary hepatocytes were determined by Western blotting .RESULTS:Incubation with 35 mmol/L glucose re-sulted in a significant decrease in cell viability and an increase in apoptosis , whereas mannitol had no significant effect on the cell viability or apoptosis .A progressive depolarization of the mitochondria , an increase in cytosol cytochrome C and a dramatic decrease in mitochondrial cytochrome C in high-glucose stressed hepatocytes were observed .The enzymatic activi-ties of caspase-9 and caspase-3, but not caspase-8, were significantly increased in high glucose-stressed hepatocytes ( P<0.05).High glucose treatment suppressed the expression of Bcl-2 and Bcl-xL, while it increased the expression of the pro-apoptotic factor Bax .CONCLUSION:High glucose stress reduces mitochondrial membrane potential , initiates mitochon-dria-mediated apoptotic pathways and promotes apoptosis of hepatocytes with steatosis .This may be an important pathologi-cal mechanism of hyperglycemia-induced progression of nonalcoholic fatty liver disease .

BACKGROUND: Intra-articular injection of hormone has been applied in the treatment of arthritis because it can alleviate arthralgia rapidly, which is accompanied commonly by progressive cartilage impairments. It is not clear if supplement of growth factor like insulin effect can play a protective role in articular chondrocytes.OBJECTIVE: To observe the effects of insulin or hydrocortisone alone and the combination on the proliferation of chondrocytes.DESIGN: Grouping comparative study, the effect of one medicine was analyzed by using one-factor analysis of variance, while the combined effect was analyzed with multi-factor analysis of variance.SETTING: Guangdong Institute of Trauma Sugery.MATERIALS: Articular cartilage from the knees of New Zealand white rabbits of 4 - 6 weeks old.METHODS: This study was carried out at Guangzhou Traumatic Research Institute from Feberary 2000 to May 2001. Chondrocytes were isolated from the knee joints of New Zealand white rabbits, digested with hyaluronidase,pancreatin and type Ⅱ collagenase and exposed to insulin, hydrocortisone or the combination of insulin and hydrocortisone of different dosage. They were divided into four groups:Control group ( without adding insulin and hydrocortisone), insulin group (0. 035,0. 35,3.5,35 mg/L subgroups), hydrocortisone group(1,5,10,50,100 mg/L subgroups) and insulin(0. 35 mg/L) combined with hydrocortisone(50 mg/L) group. Their influence on chondrocytes proliferation was observed with methyl thiazolyl tetrazolium(MTT) method.sulin.at the concentration of 0. 035 mg/L( P ＜ 0.01 ), reaching the maximum at could inhibit the proliferation of chondrocytes ( P ＜ 0.05 ), which became significant with increasing concentration and no viable chondrocytes could be exposed to 0 . 35 mg/L insulin combined with 50 mg/L hydrocortisone, the promoting effect of insulin was inhibited due to negative cooperation.CONCLUSION: Insulin at low concentration could enhance the proliferation of chondrocytes, but hydrocortisone displayed inhibiting effect on the growth of chondrocytes. The function of insulin was antagonized when combined with hydrocortisone.

BACKGROUND: Type Ⅱ collagen has been used as the carrier for chondrocyte transplantation in animal models, but whether type Ⅱ collagen may cause arthritis or mediate cytotoxicity remains unknown.OBJECTIVE: To detect the cellular immune functions of the New Zealand rabbits immunized by porcine type Ⅱ collagen.DESIGN: An exploratory comparative study based on the observations.SETTING: An institute of trauma surgery of a municipal hospital.MATERIALS: The study was conducted in the Institute of Trauma Surgery,Guangzhou Red Cross Hospital from August 1999 to February 2000. Six New Zealand rabbits, whose body mass ranged from 2.0 kg to 3.0 kg, were chosen of either gender.METHODS: The rabbits were immunized by porcine type Ⅱ collagen for 60days, during which the plasma was regularly taken for detection of type Ⅱ collagen antibody. On the 60th day, the peripheral blood as well as the spleens and lymph nodes were taken to separate the lymphocytes, which were subjected to secondary stimulation with type Ⅱ collagen in vitro to observe the reactive cell proliferation. The lymphocytes were randomly divided into two groups, and the first group was treated with phytohemagglutinin(PHA) of different concentrations to serve as the positive control, in which non-specific immunity was examined; The second group was treated with type Ⅱ collagen of different concentrations for examining specific immunity.peripheral blood lymphocytes of normal and immunized rabbits.RESULTS: On the 21st day, the titer of the antibody presented the first peak, and 40 days after the re-injection of the antigen the second peak appeared, which maintained for 20 days and then gradually descended. The lymphocytes of the normal rabbits proliferated in response to PHA stimulation but not to the first stimulation with the type Ⅱ collagen. The lymphocytes of the immunized rabbits exhibited significant proliferation upon stimulations with both PHA and type Ⅱ collagen. At the concentration of 25 mg/L, type Ⅱ collagen stimulation was sufficient to induce lymphocyte proliferation, the peak of which occurred when the collagen concentration reached 50 mg/L.CONCLUSION: Xenogenic type Ⅱ collagen at an adequate concentration may induce the increase of the type Ⅱ collagen antibody in immunized rabbits and proliferation of lymphocytes of the spleens and peripheral blood to cause cellular immune reaction and even immunological arthritis in relation to the transplantation.

Objective To investigate the relationship between various stages of chronic hepatitis B virus (HBV) infection and lipid metabolism and its influencing factors.Methods Seventy-two cases of chronic hepatitis B (CHB),40 cases of liver cirrhosis and 17 cases of hepatocellular carcinoma (HCC) were enrolled.One-way ANOVA analyses were used to compare age,gender,liver function,lipid metabolism,and HBV DNA levels of each group.Pearson correlation analysis was used to study the correlation between HBV DNA and lipid metabolism.Binary Logistic regression analyses were performed to explore the risk factors of cirrhosis and HCC in patients with CHB.Results Differences of age,alanine aminotransferase (ALT),albumin (Alb),triglyceride (TG),and cholesterol(CHO) among the three groups (CHB group,cirrhosis group and HCC group) were statistically significant (all P＜0.05).TG levels in cirrhosis and HCC groups were (-0.061± 0.234)lg mmol/L and (-0.061±0.253) lg mmol/L,respectively,which were both significantly lower than that of the CHB group (0.116±0.182) lg mmol/L (F=11.466,P=0.000).CHO level in cirrhosis group was (0.460±0.333) lg mmol/L,which was lower than that in CHB group (0.586±0.101) lg mmol/L (F=4.892,P=0.009).The HBV DNA levels inversely correlated with TG and CHO levels in CHB group (r=-0.266,P=0.024; r=-0.309,P=0.008,respectively).The HBV DNA levels of cirrhosis and HCC patients positively correlated with ALT levels (r=0.355,P =0.007).Old age (OR=1.096,95％CI:1.025-1.172),low Alb (OR=0.000,95％CI:0.000-0.000),and low levels of ALT (OR=0.128,95％CI:0.026-0.641) were risk factors for development of cirrhosis and HCC in CHB patients (all P＜0.05).Conclusions With the progression of liver injuries,TG and CHO levels are reduced.Further studies of correlation between risk factors for the development of cirrhosis and HCC and lipid metabolism in CHB patients are needed.

BACKGROUND:Thoracic spinal cord injury often leads to double lower limb paralysis. Paraplegia walking orthosis can improve lower limb dysfunction, improve the daily living activity, and regain the ability to stand and walk in patients with paraplegia. OBJECTIVE:To discuss the effects of paraplegia walking orthosis on muscle spasticity and recovery of function of the affected lower extremity in patients with thoracic spinal cord injury. METHODS:The 20 patients with thoracic spinal cord injury (T5-12), according to the damage plane by American Spinal Injury Association standard, were divided into complete damage group and incomplete damage group (n=10). Al patients were fitted out paraplegia walking orthosis. They received residual muscle strength training, sitting balance training, and transfer training prior to assembly, and then subjected to standing exercise within paralel bar, balance and transfer training, and walking aid devices training indoor and outdoor, and elbow crutch training on foot after the assembly. RESULTS AND CONCLUSION:Compared with pre-treatment, American Spinal Injury Association score increased at 12 weeks after treatment with paraplegia walking orthosis, and sensation did not obviously alter. Spasm worsened with prolonged course of disease in the complete damage group. At 12 weeks after treatment, American Spinal Injury Association score increased, sensation apparently improved, and the spasm did not change with time in the incomplete damage group. Activities of daily living (modified Barthel index, and functional independence evaluation) evidently improved in both groups. Compared with 2 weeks, the 10-m walking time was noticeably reduced and the 6-minute walking distance was prolonged at 12 weeks in both groups. These results confirm that paraplegia walking orthosis fitted out in patients with thoracic spinal cord injury significantly improves the patient’s motor function, activities of daily living and walking ability, and also has certain influence on muscle spasm control.

OBJECTIVE To study the audiological and etiological characteristics of infants failed to pass hearing screening. METHODS 126 infants received audiological diagnostic tests,including auditory brainstem response(ABR),40 Hz auditory event related potential(40 Hz AERP),distortion product otoacoustic emissions(DPOAE),tympanometry and acoustic reflex. The degrees and types of the hearing loss,and etiological characteristics were analyzed. RESULTS Among 126 infants (252 ears),61 were diagnosed with sensorineural hearing loss(48.41%),48 were conductive hearing loss(38.09%),and 17 were found to have normal ABR thresholds(13.49%). The hearing loss was associated with various factors,including history of infection during pregnancy(21 cases),threatened abortion(9 cases),pregnancy with age at or over 35(6 cases),extension of pregnancy(7 cases),history of systematic diseases(10 cases),history of neonatal jaundice(13 cases),history of asphyxia and hypoxia(18 cases),premature and low birth weight neonates(8 cases),neonatal diseases (8 cases),family history of deafness(5 cases),craniofacial deformity(3 cases),central nervous system disorder(6 cases),and 9 cases were second child. CONCLUSION The infants who failed to pass hearing screening have various etiology characteristics in hearing loss. The infants associated with risk factors were mostly found to have sensorineural hearing loss.

Regulation of the adhesion molecules expression by cytokine in vascular endothelial cells was investigated. Human umbilical vein endothelial cells (HUVEC) were stimulated with cytokines, TNF-α (1-250 U/ml) or IL-1β (0.1-50 U/ml) for 24 h. HUVEC were also cultured with cytokines, TNF-α (100 U/ml) or IL-1β (10 U/ml), for 4-72 h, cell surface expression of adhesion molecules (ICAM-1 and VCAM-1) were detected and quantitated by immunocytochemical methods and computerized imaging analysis technique. Adhesion molecules expression were up-regulated by TNF-α, IL-1β in a concentration- and time-dependent manner. Some significant differences were observed between the effects of cytokines on the ICAM-1 and on VCAM-1 expression. Cytokines might directly induce the expression of ICAM-1 and VCAM-1 in vascular endothelial cells. Our observations indicate differential functions of the two adhesion molecules during the evolution of inflammatory responses in stroke.

Regulation of the adhesion molecules expression by cytokine in vascular endothelial cells was investigated. Human umbilical vein endothelial cells (HUVEC) were stimulated with cytokines, TNF-α (1-250 U/ml) or IL-1β (0.1-50 U/ml) for 24 h. HUVEC were also cultured with cytokines, TNF-α (100 U/ml) or IL-1β (10 U/ml), for 4-72 h, cell surface expression of adhesion molecules (ICAM-1 and VCAM-1) were detected and quantitated by immunocytochemical methods and computerized imaging analysis technique. Adhesion molecules expression were up-regulated by TNF-α, IL-1β in a concentration- and time-dependent manner. Some significant differences were observed between the effects of cytokines on the ICAM-1 and on VCAM-1 expression. Cytokines might directly induce the expression of ICAM-1 and VCAM-1 in vascular endothelial cells. Our observations indicate differential functions of the two adhesion molecules during the evolution of inflammatory responses in stroke.