[Investigation of a cluster epidemic of COVID-19 in Ningbo].

Objective: To investigate a cluster epidemic of COVID-19 after a mass gathering activity in Ningbo of Zhejiang province and analyze the transmission chain and status of infection cases of different generations. Methods: The tracking of all the close contacts of the first COVID-19 case and epidemiological investigation were conducted on January 29, 2020 after a cluster epidemic of COVID-19 related with a Buddhism rally on January 19 (the 1.19 rally) in Ningbo occurred. The swabs of nose/throat of the cases and close contacts were collected and tested for nucleic acids by real-time fluorescence quantitative RT-PCR. Results: From January 26 to February 20, 2020, a total of 67 COVID-19 cases and 15 asymptomatic infection cases related with the 1.19 rally were reported in Ningbo. The initial case was the infection source who infected 29 second generation cases and 4 asymptomatic infection cases, in whom 23 second generation cases and 3 asymptomatic infection cases once took bus with the initial case, the attack rate was 33.82% (23/68) and the infection rate was 38.24% (26/68). The risks of suffering from COVID-19 and being infected were 28.91 times and 26.01 times higher in rally participants taking bus with initial case compared with those taking no bus with initial case. In this epidemic, 37 third+generation cases and 11 related asymptomatic infection cases occurred, the attack rate was 2.88% (37/1 283) and the infection rate was 4.76% (48/1 008). The main transmission routes included vehicle sharing and family transmission. Conclusion: It was a cluster epidemic of COVID-19 caused by a super spreader in a massive rally. The epidemic has been under effective control.

[Study on transmission dynamic of 15 clusters of COVID-2019 cases in Ningbo].

Objective: To describe the basic characteristics of clusters of coronavirus diseases 2019 (COVID-19) cases in Ningbo, Zhejiang province, and evaluate the generation time (Tg) and basic reproduction number (R(0)) of COVID-19. Methods: The basic information and onset times of the clusters of COVID-19 cases in Ningbo were investigated, the inter-generational interval of the cases were fitted by using gamma distribution, and the R(0) was calculated based on the SEIR model. Results: In the 15 clusters of COVID-19 cases, a total of 52 confirmed cases, 5 cases of nucleic acid-positive asymptomatic cases. The cases occurred from January 23 to February 4, the cases were mainly women. The incubation period was (6.11±3.38) days, and the median was 5 days. The Tg was (6.93±3.70) days. There were no significant differences in Tg between age group<60 years and age group 60 years and above, and between men and women (P=0.551). According to the Tg calculated in this paper, the R(0) of COVID-19 in Ningbo was 3.06 (95%CI: 2.64- 3.51); according to the reported case transmission interval of 7.5 days in the literature, the R(0) was 3.32 (95%CI: 2.51-9.38). Conclusion: There is no age and gender specific differences in the Tg of clusters of COVID-19 cases in Ningbo, and COVID-19 has high infectivity and spreading power in early phase.

[Newborn screening program and blood amino acid profiling in early neonates with citrin deficiency].

Objective: To investigate the profiles of blood amino acid and acylcarnitine in early neonates with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) and the sensitivity of newborn screening, and to explore potential biochemical metabolic markers for newborn screening program. Methods: Amino acid and acylcarnitine profiles in dried blood spots of newborn screening program were analyzed by tandem mass spectrometry (MS/MS). A total of 158 651 neonates born in Guangzhou from January 1, 2015 to June 30, 2019 were enrolled in this newborn screening program, and additionally 55 patients with NICCD confirmed by SLC25A13 gene analysis in Guangzhou Women and Children Medical Center were included in this study. NICCD screen-positive was defined as the cutoff value of citrulline (Cit) ≥ 30 µmol/L. The values of blood sampling time of the true positive group and those of the false negative group were compared by t-test. The levels of amino acid and acylcarnitine among different groups, including true positive group (Cit≥30 µmol/L), false negative group (Cit 21-<30 µmol/L and Cit<21 µmol/L) and the normal control group, were analyzed by F test, respectively. Results: Among 158 651 neonates, 39 neonates were positive for NICCD screening. Three of them were confirmed NICCD and 4 cases were found to be false negatives. The positive predictive value was 7.7% and the sensitivity was about 43.0%. Among 55 patients with NICCD, 18 cases (18/55, 32.7%) were true positives and 37 cases (37/55, 67.3%) were false negatives based on the cutoff value of citrulline in the dried blood spots for newborn screening. The blood sampling time was significantly different between true positive group and false negative group ((4.28±1.6) vs. (2.98±0.74) d, t=4.06, P<0.01). The increased levels of tyrosine((176.0±98.4) µmol/L), methionine ((37.0±26.9) µmol/L) and phenylalanine ((133.0±80.9)µmol/L) in Cit≥30 µmol/L group (n=18) were significantly different as compared with those in the other three groups, respectively (F=117.0, 58.5, 135.0, P<0.01). The levels of arginine ( (10.0±9.2) , (11.0±9.3) , (9.0±17.8) µmol/L), valine ( (119.0±29.8) , (107.6±14.1) , (102±68) µmol/L) and leucine ( (167.0±37.1) , (161.0±37.7) , (163.5±180.6) µmol/L) were not statistically significant among groups of Cit≥30 µmol/L(n=18), Cit21-<30 µmol/L(n=7) and Cit<21µmol/L(n=30,P>0.05), but they were significantly higher than those of the normal control group ((4±3), (78±21), (114.0±31.5) µmol/L, n=1 000), respectively(F=30.1, 23.0, 29.8, P<0.01). Alanine (Ala) ( (150±50) , (156.0±30.2), (168±105), (152±52) µmol/L) levels showed no significant difference (F=0.86, P>0.05) but the ratios of Ala/Cit (1.52±1.44, 6.82±1.56, 12.06±7.71, 19.42±6.27) decreased significantly among the four groups (F=69.0, P<0.05). The acylcarnitine levels showed no statistically significant results among the different groups (P>0.05). With Cit≥30 µmol/L and Ala/Cit<7.5 as cutoff values, the number of screen-positive cases reduced from 39 to 22 cases with no additional false negative case. With Cit≥21 µmol/L and Ala/Cit<7.5 as cutoff values the number of screen-positive cases increased to 117 cases with 1 additional true positive. Conclusions: The profiles of blood amino acid in early neonates with NICCD present the increased levels of multiple amino acids including citrulline, tyrosine, methionine and phenylalanine, and decreased ratio of Ala/Cit. Taking citrulline and ratio of Ala/Cit as screening markers can improve the positive predictive value appropriately. The limited sensitivity of NICCD newborn screening may be related to early blood sampling time.

Distribution and morphology of ghrelin immunostained cells in the adrenal gland of the African ostrich.

Ghrelin is the endogenous ligand for the growth hormone secretagogue receptor. We investigated the distribution and morphological characteristics of ghrelin-immunopositive (ghrelin-ip) cells in the African ostrich adrenal gland. We found that the adrenal gland of the African ostrich consisted of three parts: capsule, inter-renal tissue and chromaffin cells. The inter-renal tissue and chromaffin cells interdigitated irregularly. The inter-renal tissue consisted of a peripheral zone and a central inner zone. The peripheral zone could be divided into an outer subcapsular zone and an inner zone. The subcapsular zone cells were arranged as a bow, while the inner area cells formed cords that were perpendicular to the capsule. The central inner zone exhibited irregular clumps and the cells were morphologically similar to chromaffin cells. Ghrelin-ip cells were located throughout the adrenal gland except the capsule. The majority of ghrelin-ip cells were found among the chromaffin cells. The number of ghrelin-ip cells in the inter-renal tissue decreased gradually from the central inner zone, to the inner zone to the subcapsular zone. The ghrelin-ip cells were oval or irregular in shape and exhibited cytoplasmic staining. Our findings suggest that ghrelin may play a role in regulating adrenal hormone secretion in the African ostrich.

Distribution and developmental changes of ghrelin-immunopositive cells in the pancreas of African ostrich chicks (Struthio camelus).

Ghrelin, the endogenous ligand for the growth hormone secretagogue receptor (GHS-R), is produced by multiple cell types and affects feeding behavior, metabolic regulation, and energy balance. In the mammalian pancreas, the types of endocrine cells that are immunoreactive to ghrelin vary. However, little was known about its distribution and developmental changes in the pancreas of African ostrich chicks (Struthio camelus). In the present study, the distribution, morphological characteristics, and developmental changes of ghrelin-immunopositive (ghrelin-ip) cells in the pancreas of African ostrich chicks were investigated using immunohistochemistry. Ghrelin-ip cells were found in both the pancreatic islets and acinar cell regions. The greatest number of ghrelin-ip cells were found in the pancreatic islets, and were primarily observed at the periphery of the islets; some ghrelin-ip cells were also located in the central portion of the pancreatic islets. Interestingly, from postnatal d 1 to d 90, there was a steady decrease in the number of ghrelin-ip cells in the pancreatic islets and acinar cell regions. These results clearly demonstrated that ghrelin-ip cells exist and decreased with age in the African ostrich pancreas from postnatal d 1 to d90. Thus, these findings indicated that ghrelin may be involved in the development of the pancreas in the African ostrich.

Developmental morphology study on the stomach of African ostrich chicks.

This study investigates the developmental morphology changes on the proventriculus and gizzard of African ostrich chicks using gross anatomy, and light and transmission electronic microscopy. The body weight; the weight of the proventriculus and gizzard; the thickness of simple tubular glands, muscularis mucosae, compound tubular glands, and muscular layer of the proventriculus; and the thickness of the gizzard glands and muscularis mucosae were measured on postnatal d one, 45, 90, and 334. Under transmission electronic microscopy, the oxynticopeptic cells, mucous cells, and endocrine cells were observed in the proventriculus on postnatal d one, 45, and 90. Our results revealed that the weights of the proventriculus and gizzard (relative to the body weight) both peaked on d 90, respectively, and declined thereafter. The thicknesses of the simple tubular glands, muscularis mucosae, compound tubular glands, muscular layer of the proventriculus, and gizzard glands increased with increasing bird age. However, the thickness of the muscularis mucosae in the gizzard peaked on d 90 and gradually decreased thereafter. The result of transmission electron microscopy revealed that the glands in the submucosa layer of the proventriculus did not differ from other cell types, aside from an increase in cell diameter and an increase in cytoplasmic content. A significant increase in the number of mitochondria and smooth endoplasmic reticulum was noted, as well as an in granules secreted by endocrine cells. Therefore, to improve brood rates of reared African ostrich chicks, feed management ought to be enhanced between postnatal d one and day 90.

[Association between IKZF3 gene polymorphisms and systemic lupus erythematosus in Han ethnic group in southern China: a case-control study].

OBJECTIVE: To understand the association between IKZF3 gene polymorphism and the risk of systemic lupus erythematosus(SLE)in Han ethnic group in southern China. METHODS: A case-control study was conducted among 213 SLE patients and 234 healthy controls. Venous blood samples were collected from them to measure single nucleotide polymorphism(SNP)in IKZF3 by using the method of restriction fragment length polymorphism(PCR-RFLP). Multivariate logistic analysis and generalized multifactor dimensionality reduction(GMDR)method were used under multiple genetic models(additive, dominant, recessive), to analyze the association between IKZF3 and SLE susceptibility or different clinical features and gene-gene interactions. In addition, bioinformatics analysis was also conducted. RESULTS: As for rs114509391, CA genotype might decrease the risk of SLE compared with AA genotype(OR=0.14, 95%CI: 0.03-0.56, P=0.006)and significant association was also observed under dominant model(OR=0.26, 95%CI: 0.09-0.81, P=0.02). Stratified analysis indicated that rs9635726 and rs9909593 were related to SLE onset. The study of clinical features showed that rs907091 was associated with both renal disorder(additive: OR=0.59, 95%CI: 0.35-0.98, P=0.043)and anti-SSB(dominant: OR=0.41, 95%CI: 0.18-0.96, P=0.040). rs9635726 GG and GA genotype might decrease the risk of anti-SSB compared with AA genotype(OR=0.37, 95%CI:0.16-0.88, P=0.025). In addition, bioinformatics analysis indicated that all the studied SNPs were functional. CONCLUSION: IKZF3 rs114509391, rs9635726 and rs9909593 polymorphisms might be related to SLE susceptibility in Han ethnic group in southern China and rs9909593, rs907091 might be associated with renal disorder and anti-SSB.

[A meta-analysis of microRNA-149, microRNA-499 gene polymorphism and susceptibility to hepatocellular carcinoma].

OBJECTIVE: To investigate the relationship between microRNA-149 (rs2292832), microRNA-499 (rs2292832) polymorphism and hepatocellular carcinoma susceptibility by meta-analysis. METHODS: We used"hepatocellular carcinoma/HCC","miRNA-149/miR-149/microRNA-149", and"miRNA-499/miR-499/microRNA-499"as key words to search papers in databases including China National Knowledge Internet (CNKI), Chinese BioMedical Literature (CBM), Vip Citation Databases (VIP), Wanfang, PubMed and Web of Science databases, and collected the case-control studies on the association of rs2292832 or rs3746444 and the susceptibility to hepatocellular carcinoma from updated to May 31st 2015. Data were extracted by two independent reviewers and pooled OR with 95% CI was calculated. A bioinformatics analysis was further conducted. RESULTS: A total of 13 research papers were collected, and 5 studies for rs2292832 and 12 studies for rs3746444. 1 096 cases and 1 701 controls were included for rs2292832 and 3 117 cases and 4 126 controls were included for rs3746444. Meta-analysis failed to detect associations between rs2292832, rs3746444 and susceptibility to hepatocellular carcinoma under each genetic model tested and alleles of OR(95% CI) were 0.99(0.78-1.28) and 1.11(0.88-1.40). However, subgroup analysis showed that rs3746444 C allele seem to be associated with an increased hepatocellular carcinoma risk in both researches which had more than 400 samples and which used more accurate genotyping methods, and OR(95%CI) were 1.32(1.02-1.70) and 1.34(1.09-1.66), respectively. Furthermore, bioinformatics analysis also showed that the expression of both SNPs were down-regulated in HepG2 cells and indicated possible functional effects on gene transcription. Cochran's Q test indicated that there was the heterogeneity among the studies included. CONCLUSIONS: No significant association was found between rs2292832, rs3746444 and susceptibility to hepatocellular carcinoma, but subgroup study indicated C allele might be associated with increased hepatocellular carcinoma risk for rs3746444. Bioinformatics analysis indicated that the two SNPs might have possible influence on gene transcription.

Association of the interleukin-6 polymorphisms with systemic lupus erythematosus: a meta-analysis.

BACKGROUND: Interleukin (IL)-6, an important proinflammatory cytokine, plays a potential pathological role in systemic lupus erythematosus (SLE). Studies on the relationship of IL-6 gene polymorphisms with SLE are inconclusive. The aim of this study was to estimate the relationship more precisely. METHODS: The databases of PubMed and Web of Science updated to 30 August 2014 were retrieved. Meta-analysis was conducted using allelic contrast, dominant, recessive and homozygote contrast models. Fifteen studies were included in this study and ethnicity-specific meta-analysis was performed on European, Iranian and Asian populations. RESULTS: Analysis for the IL-6-174 G/C polymorphism under all models except the homozygote contrast model indicated an association in the overall population (allelic contrast model: odds ratio (OR) 1.428, 95% confidence interval (CI) 1.124-1.812, dominant model: OR 1.382, 95% CI 1.037-1.842, recessive model: OR 1.610, 95% CI 1.158-2.240, homozygote contrast model: OR 1.759, 95% CI 0.989-3.127), as well as in European individuals under all four genetic models (allelic contrast model: OR 1.557, 95% CI 1.155-2.098, dominant model: OR 1.699, 95% CI 1.203-2.400, recessive model: OR 1.506, 95% CI 1.176-1.930, homozygote contrast model: OR 2.118, 95% CI 1.103-4.065). Analysis for the IL-6-572 G/C polymorphism indicated significant association in overall ethnicities under the recessive model (OR 1.491, 95% CI 1.104-2.014), but not under other models or in Asian individuals. In addition, significant association between the IL-6-174 G/C polymorphism and discoid skin lesions and antinuclear antibodies (ANAs) were found under the allelic contrast model and recessive model, respectively (discoid skin lesions: OR 2.271, 95% CI 1.053-4.895; ANAs: OR 2.244, 95% CI 1.141-4.416). CONCLUSION: This meta-analysis provides evidence of the association between the IL-6 polymorphism and the risk of SLE, hinting that the IL-6-174 G/C and IL-6-572 G/C polymorphisms may play a role in SLE susceptibility.

Netrin-1 rescues neuron loss by attenuating secondary apoptosis in ipsilateral thalamic nucleus following focal cerebral infarction in hypertensive rats.

Neurological deficit following cerebral infarction correlates with not only primary injury, but also secondary neuronal apoptosis in remote loci connected to the infarction. Netrin-1 is crucial for axonal guidance by interacting with its receptors, deleted in colorectal cancer (DCC) and uncoordinated gene 5H (UNC5H). DCC and UNC5H are also dependence receptors inducing cell apoptosis when unbound by netrin-1. The present study is to investigate the role of netrin-1 and its receptors in ipsilateral ventroposterior thalamic nucleus (VPN) injury secondary to stroke in hypertensive rats. Renovascular hypertensive Sprague-Dawley rats underwent middle cerebral artery occlusion (MCAO). Continuous intracerebroventricular infusion of netrin-1 (600 ng/d for 7 days) or vehicle (IgG/Fc) was given 24h after MCAO. Neurological function was evaluated by postural reflex 8 and 14 days after MCAO. Then, immunoreactivity was determined in the ipsilateral VPN for NeuN, glial fibrillary acidic protein, netrin-1 and its receptors (DCC and UNC5H2), apoptosis was detected with Terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP-biotin nick-end labeling (TUNEL) assay, and the expressions of caspase-3, netrin-1, DCC, and UNC5H2 were quantified by western blot analysis. MCAO resulted in the impaired postural reflex after 8 and 14 days, with decreased NeuN marked neurons and increased TUNEL-positive cells, as well as an up-regulation in the levels of cleaved caspase-3 and UNC5H2 protein in the ipsilateral VPN, without significant change in DCC or netrin-1 expression. By exogenous netrin-1 infusion, the number of neurons was increased in the ipsilateral VPN, and both TUNEL-positive cell number and caspase-3 protein level were reduced, while UNC5H2 expression remained unaffected, simultaneously, the impairment of postural reflex was improved. Taken together, the present study indicates that exogenous netrin-1 could rescue neuron loss by attenuating secondary apoptosis in the ipsilateral VPN after focal cerebral infarction, possibly via its receptor UNC5H2, suggesting that relative insufficiency of endogenous netrin-1 be an underlying mechanism of secondary injury in the VPN post stroke.

Benchmark dose approach for low-level lead induced haematogenesis inhibition and associations of childhood intelligences with ALAD activity and ALA levels.

Lead (Pb) levels, delta-aminolevulinic acid dehydratase (ALAD) activities, zinc protoporphyrin (ZPP) levels in blood, and urinary delta-aminolevulinic acid (ALA) and coproporphyrin (CP) concentrations were measured for 318 environmental Pb exposed children recruited from an area of southeast China. The mean of blood lead (PbB) levels was 75.0µg/L among all subjects. Benchmark dose (BMD) method was conducted to present a lower PbB BMD (lower bound of BMD) of 32.4µg/L (22.7) based on ALAD activity than those based on the other three haematological indices, corresponding to a benchmark response of 1%. Childhood intelligence degrees were not associated significantly with ALAD activities or ALA levels. It was concluded that blood ALAD activity is a sensitive indicator of early haematological damage due to low-level Pb exposures for children.

Adverse health effects of lead exposure on children and exploration to internal lead indicator.

Our research on adverse effects of lead exposures on physical and neurobehavioral health of children aged 6-12years in 4 villages, labeled as K, M, L, and X, in rural China, was reported in this article. Lead in blood (PbB), urine (PbU), hairs (PbH), and nails (PbN) were measured by graphite furnace atomic absorption spectrometry. Abbreviated Symptom Questionnaire of Conner's instruments and Revised Raven's Standard Progressive Matrices were applied to evaluate childhood attention deficit/hyperactivity disorders (ADHD) and intelligences. Geometric means (SD) of PbB, PbU, PbH and PbN concentrations were 71.2 microg/L (1.56), 11.7 microg/g (1.75), 12.5 microg/g (2.82), and 25.3 microg/g (2.79), respectively. 54 (17.0%) children had PbB levels of > or =100 microg/L. Boys, 6-10 years old, and living in village K were 2.11, 2.48, and 9.16 times, respectively, more likely to be poisoned by lead than girls, aged 11-12 years, and residing in X. 18 (5.7%) and 37 (11.7%) subjects had ADHD and mental retardations, respectively. Inverse relationships between intelligences and natural log transformed PbU and PbH levels were observed with respective odds ratios (95%CI) of 1.79 (1.00-3.22) and 1.46 (1.06-2.03) or 1.28 (1.04-1.58) and 1.73 (1.18-2.52) by binary or ordinal logistic regression modeling. ADHD prevalence was different by gender and age of subjects. PbU, PbH, and PbN related to PbB positively with respective correlation coefficients of 0.530, 0.477, and 0.181. Receiver operating characteristic (ROC) curves of the three measurements revealed areas under curves (AUCs) being 0.829, 0.758, and 0.687, respectively. In conclusion, children had moderate levels of lead exposures in this rural area. Intelligence declines were associated with internal lead levels among children. ROC analysis suggests PbU an internal lead indicator close to PbB.

Correlation analysis on total lymphocyte count and CD4 count of HIV-infected patients.

OBJECTIVE: To determine the relationship between CD4 count and other blood indices and to explore the prediction of total lymphocyte count (TLC) for CD4 count in HIV-infected patients. METHODS: Cross-sectional study was performed for the prediction of TLC and other indices for CD4 count, and historical cohort study was performed for the TLC changes as a surrogate for CD4 changes of patients on antiretroviral therapy (ART) to further understanding the utility of TLC changes for AIDS patients' management. RESULTS: In our cross-sectional study, both TLC and white blood corpuscle count positively correlated to CD4 count, but differed in these patients. For patients on ART, the prediction of TLC for CD4 count is better than that of patient without ART. Further investigation of historical cohort study indicated that, among AIDS patients on highly active antiretroviral therapy, their TLC and haemoglobin changes also positively correlated to CD4 change, with a total correlation coefficient of 0.31 (p < 0.01) and 0.19 (p < 0.01) respectively. The prediction of TLC change for CD4 change differed each time point when patients underwent ART. CONCLUSIONS: Total lymphocyte count and its change can be used as alternative in conjunction with other indices to CD4 count and its change in the management of HIV-infected individuals in China.

Purification and characterization of a feline hepatic insulin receptor.

OBJECTIVE: To elucidate the functional characteristics of a highly purified soluble liver insulin receptor in cats. SAMPLE POPULATION: Frozen livers from domestic cats were obtained commercially. PROCEDURES: The feline hepatic insulin receptor was purified from Triton X-100 solubilized plasma membranes by the use of several chromatography matrices, including affinity chromatography on an insulin-Sepharose matrix. RESULTS: The receptor, although not homogeneous, was purified 3,000-fold. Two silver-stained protein bands were identified following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with molecular weight of 134,000 and 97,000, which are similar to insulin receptors isolated from other animals. This isolated receptor had steady-state insulin binding by 40 minutes at 24 C. Optimal insulin binding occurred at pH 7.8 and with 150 mM NaCl. Under these conditions, a curvilinear Scatchard plot was obtained with the isolated receptor. Using a 2 binding-site model, the feline insulin receptor had a high-affinity low-capacity site with a dissociation constant (KD; nM) of 3 and a low-affinity high-capacity site with a K(D) of 1,180. The receptor also had tyrosine kinase activity toward an exogenous substrate that was stimulated by insulin and protamine. CONCLUSIONS AND CLINICAL RELEVANCE: Many of the reported characteristics of the liver insulin receptor in cats are similar to those for the receptor isolated from other animals and tissues, although some differences exist. These similarities suggest that characterization of the feline insulin receptor is important to understanding insulin resistance in cats with diabetes as well as in humans with diabetes.

Cervical spondylotic myelopathy treated by anterior multilevel decompression and fusion. Follow-up report of 214 cases.

From 1974 to 1982, 214 patients were treated for cervical spondylotic myelopathy by anterior multilevel decompression and the Robinson anterior interbody fusion and followed for an average of three years and 11 months. Postoperative results were superior to those obtained previously using posterior decompression. The average patient age was 50.1 years (range, 30-75); there were 187 men and 27 women. Overall, the percent of patients with excellent and good results was 59.3%; with fair results, 29%, no change; 7.9%, and deterioration, 3.8%. Of the 111 patients who had the disease less than one year before treatment, 74% were categorized after surgery as excellent or good. In contrast, only 36% of the 66 patients who had the disease more than two years were rated excellent and good. Myelography is valuable in diagnosing and locating lesions. The average number of discs removed was 3.1 per patient. The number of cases with excellent and good results was the same whether or not union of the graft occurred. Reoperation was required in 12 patients, and a third operation was performed in three patients. Eventually, 93% of the patients were able to lead independent lives.