[Correlation analysis of low-dose X-ray ionizing radiation and thyroid function in radiation workers].

Objective: To investigate the effect of low-dose X-ray ionizing radiation on thyroid function of radiation workers. Methods: From January to December 2021, a total of 1039 medical workers in some tertiary hospitals in Wuhan were selected as the survey subjects, of which 518 radiation workers were selected as the exposure group, and 521 non-radiation workers were selected as the control group. The general conditions of the two groups were collected, and 5 indicators of thyroid function were measured, including total thyroxine (TT(4)) , total triiodothyronine (TT(3)) , free triiodothyronine (FT(3)) , thyroid stimulating hormone (TSH) , and free thyroxine (FT(4)) . The annual cumulative dose of ionizing radiation exposure in the exposure group was collected. Pearson χ(2) test and independent sample t test were used to compare the general conditions, 5 indicators of thyroid function and abnormal rate between the two groups. Linear regression model was used to analyze the correlation between the annual cumulative dose and 5 indicators of thyroid function in the exposure group. Binary logistic regression was used to analyze the influencing factors of thyroid dysfunction in the exposure group. Results: The TT(4) levels of the workers in the control group and the exposure group were (7.95±1.07) µg/dl and (8.26±1.41) µg/dl, respectively, and the FT(4) levels were (16.33±2.19) pmol/L and (17.15±2.42) pmol/L, respectively, the rate of thyroid dysfunction was 4.80% (25/521) and 8.49% (44/518) , and the above differences were statistically significant (P<0.05) . Linear regression analysis showed that the annual cumulative dose of the exposure group was significantly correlated with TT(4), TT(3), FT(4), and TSH (P<0.05) . For every 1 mSv increase in the annual cumulative dose, TT(4) increased by 1.661 µg/dl, FT(4) increased by 1.422 pmol/L, TT(3) decreased by 0.113 ng/ml, and TSH decreased by 0.731 µIU/ml. Binary logistic regression analysis showed that the older the radiation workers, the higher the risk of thyroid dysfunction (OR=1.080, 95% CI: 1.016-1.148, P=0.013) ; the greater the annual cumulative dose, the higher the risk of thyroid dysfunction (OR=6.400, 95%CI: 1.796-22.811, P=0.004) . Conclusion: The annual cumulative dose of low-dose X-ray ionizing radiation is positively correlated with thyroid function TT(4) and FT(4) of radiation workers, and negatively correlated with TT(3) and TSH; the greater the age and annual cumulative dose, the higher the risk of thyroid dysfunction.

[Cognitive function analysis of chronic lead poisoning in adults].

Objective: To explore the changes of the auditory event-related potentials P300 and the Montreal Cognitive Assessment (MoCA) in the chronic mild lead poisoning in order to find out the impairment of cognitive function and intervene early. Methods: In February 2020, 50 patients with chronic mild lead poisoning in Wuhan Center for Prevention and Treatment of Occupational Diseases from June 2011 to June 2015 were selected as the case group, and 50 healthy people were selected as the control group. The changes of auditory event-related potential P300 and MOCA of the two groups were analyzed. Results: Compared with the control group, the latency of P300 of auditory event-related potential in the case group was prolonged and the amplitude was decreased (P<0.05) . Compared with the control group, the total score of MoCA in the case group was decreased, the mean score of language, abstract and delayed memory items decreased, and the differences were statistically significant (P<0.05) . Conclusion: The combination of auditory event-related potential P300 and MOCA is helpful to detect the early cognitive impairment in chronic lead poisoning population, and auditory event-related potential P300 is an objective and effective early detection method.

[Investigation of occupational noise exposure and hearing loss among automobile manufacturing workers].

Objective: To investigation the situation of occupational noise exposure and hearing loss among workers in automobile manufacturing enterprise during 2017-2019 in Wuhan. Methods: Workers in automobile manufacturing who underwent physical examination in Wuhan Hospital for the Prevention and Treatment of Occupational Diseases from 2017 to 2019 were included as subjects in the cross-sectional survey. Questionnaire survey, noise detection and pure tone threshold test were used. Excluding individuals with working time less than 3 years and information deficiency, 3 948 individuals were finally included in the study. Results: Among 3 948 workers, 128 workers had hearing loss and the rate of hearing loss was 3.24%, among which 101 workers had high-frequency hearing loss and 27 workers were diagnosed as occupational noise deafness. The prevalence of hearing loss among workers previously exposed to noise was significantly higher than that without prior exposure (12.10%, 0.96%, P<0.05) . The prevalence of hearing loss among workers with occupational noise exposure <80 dB (A) , 80~<85 dB (A) and ≥85 dB (A) was 1.83%, 2.69% and 5.09%, respectively. The prevalence of high frequency hearing loss was 1.60%, 2.05% and 3.71%, respectively. The prevalence of occupational noise deafness was 0.23%, 0.64% and 1.38%, respectively. The prevalence of hearing loss and high frequency hearing loss among workers exposed to different occupational noise was statistically significant (P<0.05) , while the prevalence of occupational noise deafness was not statistically significant (P>0.05) . There were statistically significant differences in the prevalence of hearing loss (2.88%, 4.45%) and occupational noise deafness (0.46%, 1.41%) between those who used protective equipment and those who did not (P<0.05) . Compared with workers exposed to occupational noise <80 dB (A) , workers exposed to occupational noise ≥85 dB (A) had A 3.16-fold increased risk of hearing loss (OR=3.16, 95%CI: 1.44~6.95, P<0.05) . Compared to workers using hearing protective equipment, the risk of hearing loss (OR=1.96, 95%CI: 1.25~3.06, P<0.05) and occupational noise deafness (OR=3.46, 95%CI: 1.51-7.96, P<0.05) significantly increased among those without using hearing protective equipment. Conclusion: The risk of hearing loss in automobile manufacturing workers is significantly associated with occupational noise exposure and the use of hearing protective equipment. Good hearing protection may reduce the risk of occupational noise-induced hearing loss and occupational noise deafness.

MiR-744 mediates the Oxaliplatin chemoresistance in colorectal cancer through inhibiting BIN1.

Colorectal cancer (CRC) is one of the most common malignant cancers worldwide. However, lacking of novel and sensitive chemotherapy revealed the major challenge to improve the survival of CRC patients. The aim of this study was to explore the effect and mechanism of miR-744 on the oxaliplatin chemoresistance in CRC. Firstly, the levels of miR-744 were elevated significantly in CRC tissues from patients with oxaliplatin administration before surgery and in oxaliplatin-resistant HCT116 cells. Then, the oxaliplatin chemoresistance was enhanced by miR-744 overexpression, while was attenuated by miR-744 inhibition in HCT116 and T84 cells. Additionally, the level of BIN1 protein was found to be regulated negatively by miR-744, and BIN1 overexpression blocked the oxaliplatin chemoresistance induced by miR-744. Furthermore, BIN1 was proved to be a direct target of miR-744 by luciferase reporter assay. Taken together, these findings demonstrated that miR-744 might positively mediate the oxaliplatin chemoresistance through suppressing BIN1 expression in CRC cells, thus suggested a rationale target for the developing more effective strategies to reverse oxaliplatin resistance in CRC treatment.

[Determination of n-butyl alcohol in urine by headspace solid-phase microextraction coupled with gas chromatography].

Objective: To establish a headspace solid phase microextraction-gas chromatography method for determination of n-Butyl alcohol in urine. Methods: In October 2019, the n-butyl alcohol in urine was extracted with a polydimethylsiloxane/divinylbenzene (PDMS/DVB) solid-phase microextraction head. The conditions of salt amount, extraction temperature, extraction time and desorption time were optimized. The separation was performed on HP-5 (30 m×0.32 mm×0.25 µm) capillary column and detected with flame ionization detector. The quantification was based on the external standard curve. Results: The linear relationship of n-butyl alcohol in urine was good in the range of 0.04-3.00 mg/L, the correlation coefficient was 0.999, the detection limit of the method was 0.04 mg/L, the recovery was 77.4%-102.8%, the intra-run precision was 3.67%-8.11%, and the inter-assay precision was 4.94%-6.90%. Conclusion: The method has simple operation, high concentration efficiency and high sensitivity, and it is suitable for the determination of n-butyl alcohol in urine of occupational exposure to n-butyl alcohol.

[Investigation of neck and shoulder pain among workers in an automobile plant].

Objective: To investigate the prevalence and risk factors of neck and shoulder pain (NSP) among automobile manufacturing workers and to provide a theoretical basis for prevention of NSP. Methods: From November 5 to November 19, 2017, a total 446 works who had worked for more than one year were recruited from an automobile plant by cluster sampling method. Chi square test and unconditional logistic regression were used to exam the relation between influencing factors and NSP. Results: The annual prevalence rate of NSP was 34.8%. Multifactor regression analysis showed that age、work fatigue、department staff shortages、lifting heavy objects in awkward positions、neck flexion foreword and prolong sitting position work were the risk factors of NSP (OR=2.18, 95%CI:1.49~3.18; OR=4.52, 95%CI:1.27~16.00; OR=1.66, 95%CI:1.04~26.68; OR=2.10, 95%CI:1.16~3.81; OR=2.25, 95%CI:1.39~3.66; OR=2.42, 95%CI:1.06~5.56) and work break was the benefit factors of NSP (OR=0.58, 95%CI:0.36~0.94) . Conclusion: The annual prevalence rate of NSP among automobile manufacturing workers was high. Lifing heavy objects、awkward working positions and unreasonable work arrangement were the major risk factors of NSP, and work break can effectively reduce the risk of NSP. Effective ergonomic intervention should be carried out to prevent the occurrence of NSP.

Chemical composition and antifungal activity of Carica papaya Linn. seed essential oil against Candida spp.

In recent years, the incidence of clinical yeast infections has increased dramatically. Due to the extensive use of broad-spectrum antifungal agents, there has been a notable increase in drug resistance among infections yeast species. As one of the most popular natural antimicrobial agents, essential oils (EOs) have attracted a lot of attention from the scientific community. The aim of this study was to analyse the chemical composition and examine the antifungal activity of the EO extracted from the seeds of Carica papaya Linn. The papaya seed EO was analysed by gas chromatography-mass spectrometry. The major constituent is benzyl isothiocyanate (99·36%). The filter paper disc diffusion method and broth dilution method were employed. The EO showed inhibitory effect against all the tested Candida strains including C. albicans, C. glabrata, C. krusei, C. parapsilosis and C. tropical with inhibition zone diameters in the range of 14·2-33·2 mm, the minimal inhibitory concentrations (MICs) in the range of 4·0-16·0 µg ml-1 and the minimum fungicidal concentrations (MFCs) in the range of 16·0-64·0 µg ml-1 . Here, we found that the papaya seed EO has promising anticandida activity and identify C. papaya L. as a potential natural source of antifungal agents. SIGNIFICANCE AND IMPACT OF THE STUDY: The chemical composition and antifungal activity of essential oil of Carica papaya seeds were studied. The oil of papaya seeds could inhibit the growth of Candida spp. for the first report. Carica Papaya may be recognized as a possible new source of natural antifungal agents.

Fermentation of Foc TR4-infected bananas and Trichoderma spp.

Fusarium wilt (also known as Panama disease) is one of the most destructive banana diseases, and greatly hampers the global production of bananas. Consequently, it has been very detrimental to the Chinese banana industry. An infected plant is one of the major causes of the spread of Fusarium wilt to nearby regions. It is essential to develop an efficient and environmentally sustainable disease control method to restrict the spread of Fusarium wilt. We isolated Trichoderma spp from the rhizosphere soil, roots, and pseudostems of banana plants that showed Fusarium wilt symptoms in the infected areas. Their cellulase activities were measured by endoglucanase activity, ß-glucosidase activity, and filter paper activity assays. Safety analyses of the Trichoderma isolates were conducted by inoculating them into banana plantlets. The antagonistic effects of the Trichoderma spp on the Fusarium pathogen Foc tropical Race 4 (Foc TR4) were tested by the dual culture technique. Four isolates that had high cellulase activity, no observable pathogenicity to banana plants, and high antagonistic capability were identified. The isolates were used to biodegrade diseased banana plants infected with GFP-tagged Foc TR4, and the compost was tested for biological control of the infectious agent; the results showed that the fermentation suppressed the incidence of wilt and killed the pathogen. This study indicates that Trichoderma isolates have the potential to eliminate the transmission of Foc TR4, and may be developed into an environmentally sustainable treatment for controlling Fusarium wilt in banana plants.

[Influence of coke oven emissions on workers' blood pressure and electrocardiographic findings].

Objective: To investigate the influence of coke oven emissions on workers' blood pressure and electrocardiographic findings, and to provide a basis for the prevention and treatment of cardiovascular diseases. Methods: The concentration of coke oven emissions at the bottom, side, and top of coke ovens was determined in a coking plant. A total of 406 coke oven workers were enrolled as exposure group and 201 office staff members were enrolled as control group. Blood pressure and electrocardiographic findings were compared between the two groups, and the multivariate logistic regression analysis was performed to analyze the influencing factors for hypertension and abnormal electrocardiographic findings. Results: The concentration of coke oven emissions was the highest at the top of coke ovens, followed by the side and bottom of coke ovens, and there was a significant difference between the exposure group and the control group (P<0.01). The exposure group had significantly higher detection rates of hypertension, abnormal electrocardiographic findings, and abnormal chest X-ray findings than the control group (P<0.05). The logistic regression analysis showed that high concentration of coke oven emission and age were risk factors for hypertension and abnormal electrocardiographic findings (P<0.05). The workers exposed to high-concentration coke oven emissions were more likely to experience hypertension and abnormal electrocardiographic findings than those exposed to low-concentration coke oven emissions (OR=1.7 and 1.9). Conclusion: Besides lung injury, coke oven emissions also have adverse effects on the cardiovascular system. Therefore, more effective measures are needed to protect the health of coke oven workers.

Indirect effect of corticotropin-releasing hormone receptor 1 gene variation on negative emotionality and alcohol use via right ventrolateral prefrontal cortex.

Variations in the corticotropin-releasing hormone receptor 1 (CRHR1) gene have been found to interact with stress in modulating excessive alcohol consumption. However, the neural mechanisms through which CRHR1 influences this risk in humans is largely unknown. This study examined the influence of an intronic CRHR1 gene variant, rs110402, on brain responses to negative emotional words, negative emotional traits, and alcohol use in adolescents and young adults at high risk for alcoholism. Childhood stress was investigated as a potential moderator. Using functional magnetic resonance imaging, we found that a region in the right ventrolateral prefrontal cortex (rVLPFC) was more engaged during negative emotional word processing in G homozygotes than in A allele carriers (p(FWE corrected) < 0.01, N = 77). Moreover, an indirect effect of genotype on negative emotionality via rVLPFC activation (p < 0.05, N = 69) was observed, which was further moderated by childhood stress (p < 0.05, N = 63). Specifically, with low childhood stress, G homozygotes exhibited lower levels of negative emotionality associated with greater rVLPFC activation, suggesting that the rVLPFC is involved in reappraisal that neutralizes negative emotional responses. In addition, we found that genotype indirectly modulated excessive alcohol consumption (p < 0.05, N = 69). Specifically, G homozygotes showed greater rVLPFC activation and had lower levels of negative emotionality, which were associated with fewer binge-drinking days and fewer alcohol related problems. This work provides support for a model in which CRHR1 gene variation modulates the risk of problem drinking via an internalizing/negative affect pathway involving rVLPFC and reappraisal of negative emotion.

Genome-wide copy number profiling using high-density SNP array in chickens.

Phenotypic diversity is a direct consequence resulting mainly from the impact of underlying genetic variation, and recent studies have shown that copy number variation (CNV) is emerging as an important contributor to both phenotypic variability and disease susceptibility. Herein, we performed a genome-wide CNV scan in 96 chickens from 12 diversified breeds, benefiting from the high-density Affymetrix 600 K SNP arrays. We identified a total of 231 autosomal CNV regions (CNVRs) encompassing 5.41 Mb of the chicken genome and corresponding to 0.59% of the autosomal sequence. The length of these CNVRs ranged from 2.6 to 586.2 kb with an average of 23.4 kb, including 130 gain, 93 loss and eight both gain and loss events. These CNVRs, especially deletions, had lower GC content and were located particularly in gene deserts. In particular, 102 CNVRs harbored 128 chicken genes, most of which were enriched in immune responses. We obtained 221 autosomal CNVRs after converting probe coordinates to Galgal3, and comparative analysis with previous studies illustrated that 153 of these CNVRs were regarded as novel events. Furthermore, qPCR assays were designed for 11 novel CNVRs, and eight (72.73%) were validated successfully. In this study, we demonstrated that the high-density 600 K SNP array can capture CNVs with higher efficiency and accuracy and highlighted the necessity of integrating multiple technologies and algorithms. Our findings provide a pioneering exploration of chicken CNVs based on a high-density SNP array, which contributes to a more comprehensive understanding of genetic variation in the chicken genome and is beneficial to unearthing potential CNVs underlying important traits of chickens.

Stimulated emission features of bound excitons in ZnO nanotubes.

We have investigated the detailed features of photoluminescence (PL) in vertically aligned ZnO nanotube (NT) arrays as a function of temperature, pumping power, and experimental geometries. In samples with different wall thickness (15 or 60 nm), the temperature-dependent PL energy followed the Varshni's formula whose fitting parameters were found to be rather close to zero-dimensional case in the 15 nm-thick NTs with much larger intensity. In reflective geometry with circular excitation beam shape, the emission gradually evolved from spontaneous to stimulated regime, inferred from amplitude and line-width variation. On the other hand, in the edge-emission geometry with needle-like excitation shape, the interaction length dependence was directly traced by using an adjustable slit.

Molecular evolutionary analysis of vertebrate transducins: a role for amino acid variation in photoreceptor deactivation.

Transducin is a heterotrimeric G protein that plays a critical role in phototransduction in the rod and cone photoreceptor cells of the vertebrate retina. Rods, highly sensitive cells that recover from photoactivation slowly, underlie dim-light vision, whereas cones are less sensitive, recover more quickly, and underlie bright-light vision. Transducin deactivation is a critical step in photoreceptor recovery and may underlie the functional distinction between rods and cones. Rods and cones possess distinct transducin α subunits, yet they share a common deactivation mechanism, the GTPase activating protein (GAP) complex. Here, we used codon models to examine patterns of sequence evolution in rod (GNAT1) and cone (GNAT2) α subunits. Our results indicate that purifying selection is the dominant force shaping GNAT1 and GNAT2 evolution, but that GNAT2 has additionally been subject to positive selection operating at multiple phylogenetic scales; phylogeny-wide analysis identified several sites in the GNAT2 helical domain as having substantially elevated dN/dS estimates, and branch-site analysis identified several nearby sites as targets of strong positive selection during early vertebrate history. Examination of aligned GNAT and GAP complex crystal structures revealed steric clashes between several positively selected sites and the deactivating GAP complex. This suggests that GNAT2 sequence variation could play an important role in adaptive evolution of the vertebrate visual system via effects on photoreceptor deactivation kinetics and provides an alternative perspective to previous work that focused instead on the effect of GAP complex concentration. Our findings thus further the understanding of the molecular biology, physiology, and evolution of vertebrate visual systems.

Intraoperative epiaortic scanning for preventing early stroke after off-pump coronary artery bypass.

BACKGROUND: Numerous studies have demonstrated the accuracy of epiaortic ultrasound scanning (EAS) for assessing ascending aortic disease. It remains unclear whether EAS changes the incidence of perioperative stroke after off-pump coronary artery bypass (OPCAB). METHODS: We studied a retrospective cohort of 2292 patients who underwent isolated OPCAB from January 2001 to December 2011. Patients were retrospectively subdivided into two groups: the non-EAS group (n=1019) who underwent OPCAB under only intraoperative transoesophageal echocardiography and the EAS group (n=1273) who underwent OPCAB under EAS. RESULTS: In the non-EAS group, 317 (31.1%) patients underwent OPCAB with partial aortic clamping and 702 (68.9%) underwent OPCAB without partial aorta clamping. In the EAS group, 301 (23.7%) patients underwent OPCAB with partial aortic clamping and 972 (76.3%) underwent OPCAB without partial aortic clamping. The incidence of early stroke was not different statistically between the EAS and non-EAS groups [non-EAS 1.7% (17/1019) vs EAS 0.8% (10/1273); P=0.052]. However, in the subgroups of patients with partial aorta clamping, the incidence of the early stroke was significantly lower in the EAS group [2.8% (9/317) vs 0.7% (2/301) P=0.041]. CONCLUSIONS: EAS has a significant clinical benefit in reducing the incidence of early stroke in cases of partial aortic clamping in OPCAB. Therefore, EAS should be considered in patients who need partial aortic clamping in OPCAB.

Whole-genome scan for signatures of recent selection reveals loci associated with important traits in White Leghorn chickens.

Chicken is considered to be an excellent model for genetic studies of phenotypic and genomic evolution, with large effective population size, specialized commercial lines, and strong human-driven selection. High-density chicken SNP chips can help to achieve a better understanding of the selection mechanisms in artificially selected populations. We performed the genome-wide tests for the selection signature in 385 White Leghorn hens and mapped positively selected regions to the genome annotations. Ten QTL related to egg production, egg quality, growth, and disease resistance traits were selected for extended haplotype homozygosity tests to give a brief overview of recent selection signatures in chicken QTL. We also reported 185 candidate genes/CDSs showing top P-values and slower decay of haplotype homozygosities. Some of these genes seemed to have significant effects on important economical traits, and most of them have not been reported in chickens. The current study provides a genome-wide map of linkage disequilibrium extents and distributions and selection footprints in the chicken genome. A panel of genes, including PRL, NCKX1, NRF1, LHX2, and SFRP1 associated with egg production, metabolism traits, and response to illumination were identified. In addition, there were more genes identified that have not yet been reported in chickens, and our results provide new clues for further study.

System design and treatment efficiency of a surface flow constructed wetland receiving runoff impacted stream water.

This study reported the efficiency of a free water surface flow constructed wetland (CW) system that receives runoff impacted stream water from a forested and agricultural watershed. Investigations were conducted to examine the potential effect of hydraulic fluctuations on the CW as a result of storm events and the changes in water quality along the flow path of the CW. Based on the results, the incoming pollutant concentrations were increased during storm events and greater at the near end of the storm than at the initial time of storm. A similar trend was observed to the concentrations exiting the CW due to the wetland being a relatively small percentage of the watershed (<0.1%) that allowed delays in runoff time during storm events. The concentrations of most pollutants were significantly reduced (p < 0.05) except for nitrate (p = 0.5). Overall, this study suggests that the design of the system could feasibly function for the retention of most pollutants during storm events as the actual water quality of the outflow was significantly better by 21-71% than the inflow and the levels of pollutants were reduced to appreciable levels.

Penalized gaussian process regression and classification for high-dimensional nonlinear data.

The model based on Gaussian process (GP) prior and a kernel covariance function can be used to fit nonlinear data with multidimensional covariates. It has been used as a flexible nonparametric approach for curve fitting, classification, clustering, and other statistical problems, and has been widely applied to deal with complex nonlinear systems in many different areas particularly in machine learning. However, it is a challenging problem when the model is used for the large-scale data sets and high-dimensional data, for example, for the meat data discussed in this article that have 100 highly correlated covariates. For such data, it suffers from large variance of parameter estimation and high predictive errors, and numerically, it suffers from unstable computation. In this article, penalized likelihood framework will be applied to the model based on GPs. Different penalties will be investigated, and their ability in application given to suit the characteristics of GP models will be discussed. The asymptotic properties will also be discussed with the relevant proofs. Several applications to real biomechanical and bioinformatics data sets will be reported.

Simultaneous inference and bias analysis for longitudinal data with covariate measurement error and missing responses.

Longitudinal data arise frequently in medical studies and it is common practice to analyze such data with generalized linear mixed models. Such models enable us to account for various types of heterogeneity, including between- and within-subjects ones. Inferential procedures complicate dramatically when missing observations or measurement error arise. In the literature, there has been considerable interest in accommodating either incompleteness or covariate measurement error under random effects models. However, there is relatively little work concerning both features simultaneously. There is a need to fill up this gap as longitudinal data do often have both characteristics. In this article, our objectives are to study simultaneous impact of missingness and covariate measurement error on inferential procedures and to develop a valid method that is both computationally feasible and theoretically valid. Simulation studies are conducted to assess the performance of the proposed method, and a real example is analyzed with the proposed method.

Inducible expression of calreticulin-N58 in Pichia pastoris by high density cell culture.

Calreticulin-N58 (CRT-N58), an active fragment of calreticulin with anti-angiogenesis activity, was expressed in P. pastoris by high density cell culture. Calreticulin-N58 DNA was synthesized by PCR and cloned to plasmid pPIC9 K resulting in the plasmid pPIC9 K-crt-N58 which was then transformed into P. pastoris GS115. The fermentation was carried out in a 50 l bioreactor with 20 l modified growth medium recommended by Invitrogen at 30°C. The cells were first grown in glycerol-PTM4 trace salts for 24 h. When the cell density was grown to A(600) = 135, methanol-PTM4 trace salts was added to induce the expression of calreticulin-N58. During the fermentation, dissolved oxygen level was maintained at 20-30%, pH was controlled at 5 by adding 7 M NH(4)OH. After 52 h of induction, the yield of secreted calreticulin-N58 was 70 mg/l and biomass growth was 293 as measured by absorption of 600 nm. The secreted calreticulin-N58 was purified to a purity of 100% by the use of SP-Sepharose FF ion-exchange chromatography (Pharmacia Biotech. NJ, USA) and desalted with ultrafiltration device (Millipore, Bedford, MA, USA). The recombinant calreticulin-N58 induced endothelial cell apoptosis and inhibited the angiogenesis on the CAM.