Inborn errors of immunity and its clinical significance in children with lymphoma in China: a single-center study

Abstract Objective: To investigate the incidence, clinical and genetic characteristics of pediatric lymphoma patients of China with inborn errors of immunity (IEI)-related gene mutations, which have not been fully studied. Method: From Jan. 2020 to Mar. 2023, IEI-related genetic mutations were retrospectively explored in 108 children with lymphomas admitted to Beijing Children's Hospital by NGS. Genetic rule and clinical characteristics as well as treatment outcomes were compared between patients with or without IEI-related gene mutations. Results: A total of 17 patients (15.7 %) harbored IEI-associated mutations, including 4 cases with X-linked lymphoproliferative syndrome (XLP), 3 cases had mutations in tumor necrosis factor receptor superfamily 13B (TNFRSF13B), 2 cases with Activated p110 syndrome (APDS). Patients with IEI all had alteration of immunocompetence with decreased levels of immunoglobulin and lymphocyte subsets. Recurrent infection existed in 41.2 % of patients. The 18-month event-free survival (EFS) and the overall response rate (ORR) of patients with IEI are significantly lower than those without IEI (33.86% vs. 73.26 %, p = 0.011; 52.94% vs. 87.91 %, p = 0.002, respectively). In addition, patients with IEI had a higher progression disease (PD) rate of 23.5 % than those without IEI of 4.4% (p = 0.006). Conclusion: The present study demonstrated that IEI-associated lymphomas were much more common than originally appreciated in pediatric lymphomas, and those were insensitive to treatment and more likely to progress or relapse. The genomic analysis and a thorough review of the medical history of IEI can be used to distinguish them from pediatric lymphomas without IEI, which are beneficial for the early diagnosis and direct intervention.

Diagnosis and treatment of 4 cases imported severe malaria and COVID-19 co-infection / 中国热带医学

@#Abstract: To explore the clinical characteristics, diagnosis and treatment of imported severe malaria and COVID-19 co-infection cases, and to provide scientific basis for epidemic prevention and control measures. The epidemiological characteristics, clinical manifestations, laboratory tests, treatment process and prognosis of 4 cases of severe malaria and COVID-19 co-infection with confirmed diagnosis were analyzed retrospectively. Four cases of severe malaria were African returnees of the same batch, male, aged 40-54 years old, with the same journey track. They all had African work and life history and acute onset. The main clinical manifestations were fever (4/4), chills (3/4), chills (3/4), nausea and vomiting (3/4), diarrhea (4/4), fatigue and anorexia (4/4). Two cases had headache and dizziness, confusion, muscle aches, two cases had cough, one cases had sputum, sore throat and runny urine. All 4 cases were confirmed by positive nucleic acid detection of the new coronavirus (2019-nCOV) in throat swabs. Plasmodium falciparum was found by microscopic examination of peripheral blood smears of all patients, and all of them were consistent with high altitude helminthiasis. All cases were accompanied by abnormal liver function and severe hypoproteinemia, two cases were hyperbilirubinemia, three cases were dyslipidemia, three cases were involved in abnormal tertiary hemogram with different degrees of elevation of procalcitonin, two cases were lactic acid poisoning, and one case was hypoglycemia. One case showed viral pneumonia on chest CT. All cases were treated individually according to the different conditions and were discharged after improvement, and were rechecked for 2019-nCOV nucleic acid and microscopic examination of blood smear negative for Plasmodium.During the global COVID-19 epidemic, the emergence of coinfection cases of con-infection of imported malaria parasites and severe acuterespiratory syndrome coronavirus 2 (SARS-CoV-2) makes the clinical diagnosis and treatment more complicated. It is important to establish the awareness of simultaneous prevention and diagnosis of COVID-19 and malaria for local prevention and control and early warning of severe cases, and timely and effective formulation of treatment plan to improve the comprehensive treatment efficiency.

Baricitinib inhibits type Ⅰ IFN-signaling during SARS-CoV-2 infection in vitro / 中国热带医学

@#Abstract: Objective　To explore the antiviral effect of baricitinib in the SARS-CoV-2 infection and influence on cytokine levels. Methods　Calu-3 cells were infected with SARS-CoV-2 at MOI of 0.1, and the levels of inflammatory cytokines (IL-6, IL-8, TNF-α and IL-1β), interferon β (IFN-β) and interferon-stimulated gene, IFIT2 in the infected cells were analyzed by qRT-PCR methods. At the same time, Calu-3 cells were infected with SARS-CoV-2 (MOI=0.1) after being treated with baricitinib for 2 hours. Cells were collected at 0, 24, 36, and 48 hours, and analyzed for the mRNA of the above genes in the drug-treated and untreated groups. Results　The mRNA levels of IL-6, TNF-a, IL-1β, IFN-β and IFIT2 in Calu-3 infected by SARS-CoV-2 cells were increased significantly. These cytokines were increased by nearly 100-fold post-infection 48 h compared with the control (P<0.000 1), and continued to increase with the infection time (P<0.001 or P<0.000 1). The increase of IL-8 mRNA level was not as significant as IL-6, TNF-α, IL-8, IL-1β, but it also showed a 2-4 folds increase. Baricitinib does not affect the level of viral RNA in Calu-3 cells after SARS-CoV-2 infection (P>0.05). However, baricitinib can significantly inhibit the up-regulation of IL-6 and TNF-α levels induced by SARS-CoV-2 infection (5.25-fold and 3.90-fold down-regulation, respectively, P<0.01), and has little effect on the levels of IL-8 and IL-1β . In addition, the drug could also significantly down-regulate the increase in IFN-β and IFIT2 levels caused by viral infection (10.51-fold and 90.78-fold down-regulation, respectively, P<0.000 1). Conclusions　Baricitinib inhibits the release of inflammatory cytokines to some extent, but it drastically down-regulates the expression of interferons and interferon-stimulated genes (ISGs), and has limited antiviral effect on SARS-CoV-2. Considering that interferon signal pathways play important roles on viral infection, caution should be exercised when using baricitinib to treat COVID-19 patients.

Ultrasonographic differentiation and Ultrasound-based management of partially cystic thyroid nodules

ABSTRACT Objective: To determine sonographic features of malignancy in partially cystic thyroid nodules and assess the diagnostic efficacy of these features for differentiating between benign and malignant lesions in the nodules with indeterminate cytology. Subjects and methods: From January 2016 to December 2017, a total of 91 patients with 94 partially cystic thyroid nodules who had undergone ultrasound-guided fine-needle aspiration biopsy and thyroid surgery in our hospital were included in this study. The sonographic features of the thyroid nodules were analyzed to identify the predictive features of malignancy and assess the diagnostic efficacy of these features. Results: The features of hypoechogenicity, microcalcification, composition, and an eccentric solid component with an acute angle had statistically significant associations with malignant nodule (p<005) by univariable analysis. Binary logistic regression analysis showed that microcalcification and hypoechogenicity were significantly associated with malignancy. Using the combination of microcalcification, hypoechogenicity, and a solid component comprising of greater than or equal to 50% of the total volume, the diagnostic sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy were 97.6%, 32.7%, 53.9%, and 94.4%, respectively. In these nodules with indeterminate cytology, this combination also exhibited a high sensitivity of 92.3% and an NPV of 83.3%. Conclusion: This study demonstrated that microcalcification and hypoechogenicity were independently associated with malignancy in partially cystic thyroid nodules. The combination of microcalcification, hypoechogenicity, and a solid portion that is greater than or equal to 50% of the total volume will help guide clinical decisions in mixed cystic solid nodules.

Identification of activated pathways in lung adenocarcinoma based on network strategy

Background: Lung adenocarcinoma has increased incidence over the past years and is the cause for almost 50% of deaths attributable to lung cancer. The objective of this paper is to identify activated pathways associated with lung adenocarcinoma based on gene co-expression network analysis. Materials and Methods: Kyoto Encyclopedia of Genes and Genomes pathway analysis of dysregulated genes was performed based on Expression Analysis Systematic Explorer test to illuminate the biological pathways. Co-expression networks of lung adenocarcinoma in different tumor Stages (IA, IB, IIA, IIB, IIIA, IIIB, and IV) were constructed by Empirical Bayes approach to reweight gene pair scores. Pathway activity analysis was conducted to compute the distribution of pathways in different stages and to identify “activated” pathways in lung adenocarcinoma. Results: We evaluated 211 dysregulated genes between lung adenocarcinoma patients and normal controls. Pathway activity analysis was performed and P values of pathways, which obtained from co-expression networks (Stage IA, IB, IIA, IIB, IIIA, IIIB, and IV), were calculated. Cell cycle, progesterone-mediated oocyte maturation, and oocyte meiosis were activated during all stages in lung adenocarcinoma. Conclusions: We successfully identified three activated pathways (cell cycle, progesterone-mediated oocyte maturation, and oocyte meiosis) in different Stages (IA, IB, IIA, IIB, IIIA, IIIB, and IV) of lung adenocarcinoma

Diversity and antimicrobial activity of culturable fungi associated with sea anemone Anthopleura xanthogrammica

Background: The main objective of this study was to isolate fungi associated with Anthopleura xanthogrammica and measure their antimicrobial and enzymatic activities. A total of 93 fungal strains associated with A. xanthogrammica were isolated in this study, of which 32 isolates were identified using both morphological characteristics and internal transcribed spacer (ITS) sequence analysis. The antibacterial activities of 32 fungal isolates were tested against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Edwardsiella tarda, Vibrio harveyi, Fusarium oxysporum, and Pyricularia oryzae by agar diffusion assay. Extracellular hydrolytic enzyme activities of the fungal isolates were determined by agar diffusion assays. Enzyme activities were detected from clear halo size. Results: The isolated fungi belonged to 18 genera within 7 taxonomic orders of 1 phylum. The genera Aspergillaceae were the most diverse and common. The antimicrobial activities of 32 isolates were evaluated, and 19 (59.4%) of fungi isolate displayed unique antimicrobial activities. All fungal strains displayed at least one enzyme activity. The most common enzyme activities in the fungi isolates were amylase and protease, while the least common were pectinase and xylanase. Conclusions: This is first report on the sea anemone-derived fungi with antimicrobial and enzyme activities. Results indicated that sea anemone is a hot spot of fungal diversity and a rich resource of bioactive natural products.

Nesiritide protects endothelial function after balloon-induced trauma in the iliac artery in rabbits

To assess the effect of nesiritide on the endothelial function of iliac arteries following endothelia trauma. Right iliac artery trauma was created with a balloon catheter. Ten rabbits were treated with a 4-week subcutaneous injection of nesiritide at a fixed daily dose of 0.1mg/kg. Ten rabbits received daily normal saline injection. Plasma endothelin 1 (ET-1), nitric oxide (NO), and Von Willebrand Factor (vWF) were measured before and after the therapies. Tissue proliferating cell nuclear antigen (PCNA) was measured after the treatment. After the treatment, in the therapeutic group, the area under internal elastic membrane and the residual lumen area were higher than in the normal saline group (P <0.05). The plasma levels of ET-1 (91.6±6.8 vs 114.9±6.3 ng/L, P =0.001), vWF (134.6±10.8% vs 188.8±10.4%, P =0.001) and the ratio of PCNA positive expression (11.7±4.2% vs 36.2±11.4%, P =0.005) in the therapeutic group was lower than in the normal saline group, while the plasma levels of NO was higher (89.7±9.3 vs 43.5±5.3 µmol/L, P =0.001). Nesiritide inhibited remodeling of rabbit iliac artery following endothelial trauma. The inhibition of vascular remodeling may be related to the alleviated endothelial dysfunction and reduced expression of tissue proliferating cell nuclear antigen

Dos mutaciones novedosas en el gen de glicina descarboxilasa en un niño con hiperglicinemia no cetósica clásica: a propósito de un caso / Two novel mutations in the glycine decarboxylase gene in a boy with classic nonketotic hyperglycinemia: case report

La hiperglicinemia no cetósica es una encefalopatía por glicina autosómica recesiva y hereditaria sumamente rara, causada por una deficiencia en el sistema enzimatico de división de la glicina mitocondrial, que provoca síntomas clínicos graves. La hiperglicinemia no cetósica se caracteriza por fenotipos diversos y complejos, por ejemplo, hipotonía, convulsiones, deterioro cognitivo, retrasos del desarrollo y espasmos mioclónicos que podrían causar apnea e incluso la muerte. En este artículo, presentamos el caso de un niño de 1 año con convulsiones mioclónicas, hipotonía y coma, con aumento de la concentración de glicina en el plasma y el líquido cefalorraquídeo y con un índice de glicina en líquido cefalorraquídeo/plasma de 0,24. Existen dos mutaciones heterocigotas novedosas que confirman el diagnóstico de hiperglicinemia no cetósica. Una es una mutación de aminoácido, c.2516A>G (p.Y839C), y la otra es una mutación en los sitios de corte y empalme, c.2457+2T>A, en el gen GLDC.

Nonketotic hyperglycinemia is an extremely rare autosomal recessively inherited glycine encephalopathy caused by a deficiency in the mitochondrial glycine cleavage system, which leads to severe clinical symptoms. Nonketotic hyperglycinemia is characterized by complex and diverse phenotypes, such as hypotonia, seizures, cognitive impairment, developmental delays and myoclonic jerks that may lead to apnea and even death. Here we report a 1-year-old boy with myoclonic seizures, hypotonia and coma; he had elevated plasma and cerebrospinal fluid glycine levels, and cerebrospinal fluid/plasma glycine ratio was 0.24. Two novel heterozygous mutations confirm the diagnosis of nonketotic hyperglycinemia. One is a missense mutation c.2516A>G (p.Y839C) and the other one is a splicing mutation c.2457+2T>A in the GLDC gene.

Risk Factors for Low Birth Weight and Preterm Birth: A Population-based Case-control Study in Wuhan, China / 华中科技大学学报（医学）（英德文版）

Low birth weight (LBW) and preterm birth (PB) are associated with newborn mortality and diseases in adulthood.We explored factors related to LBW and PB by conducting a population-based case-control study from January 2011 to December 2013 in Wuhan,China.A total of 337 LBW newborn babies,472 PB babies,and 708 babies with normal birth weights and born from term pregnancies were included in this study.Information of newborns and their parents was collected by trained investigators using questionnaires and referring to medical records.Univariate and logistic regression analyses with the stepwise selection method were used to determine the associations of related factors with LBW and PB.Results showed that maternal hypertension (OR=6.78,95％ CI:2.27-20.29,P=0.001),maternal high-risk pregnancy (OR=1.53,95％ CI:1.06-2.21,P=0.022),and maternal fruit intake ≥300 g per day during the first trimester (OR=1.70,95％ CI:1.17-2.45,P=0.005) were associated with LBW.BMI ≥24 kg/m2 of mother prior to delivery (OR=0.48,95％ CI:0.32-0.74,P=0.001) and gestation ≥37 weeks (OR=0.01,95％ CI:0.00-0.02,P＜0.034) were protective factors for LBW.Maternal hypertension (OR=3.36,95％ CI:1.26-8.98,P=0.016),maternal high-risk pregnancy (OR=4.38,95％ CI:3.26-5.88,P＜0.001),maternal meal intake of only twice per day (OR=1.88,95％ CI:1.10-3.20,P=0.021),and mother liking food with lots of aginomoto and salt (OR=1.60,95％ CI:1.02-2.51,P=0.040) were risk factors for PB.BMI ≥24 kg/m2 of mother prior to delivery (OR=0.66,95％ CI:0.47-0.93,P=0.018),distance of house from road ≥36 meters (OR=0.72,95％ CI:0.53-0.97,P=0.028),and living in rural area (OR=0.60,95％ CI:0.37-0.99,P=0.047) were protective factors for PB.Our study demonstrated some risk factors and protective factors for LBW and PB,and provided valuable information for the prevention of the conditions among newborns.

Design and implementation of digital management system in pulmonary function test room / 医疗卫生装备

Objective To develop a digital management system in the pulmonary function test room to realize pulmonary function data sharing between the outpatient department,ward,medical examination facility and community hospital.Methods The system was developed with Microsoft Visual Studio 2010 and enterprise-version SQL Server database,and the integration of pulmonary function test devices and HIS was achieved by linking up the spirometer,bluetooth height & weight digital measuring equipment,barcode scanner,back tracking system and the switch to HIS.Results The system contributed to enhancing the flow and efficiency of clinical service.Conclusion The system standardizes the flow and quality control of pulmonary function test,implements patient data sharing between the outpatient department and ward and facilitates scientific research data acquisition,and thus is worthy promoting clinically.

Design and implementation of digital management system in pulmonary function test room / 医疗卫生装备

Objective To develop a digital management system in the pulmonary function test room to realize pulmonary function data sharing between the outpatient department,ward,medical examination facility and community hospital.Methods The system was developed with Microsoft Visual Studio 2010 and enterprise-version SQL Server database,and the integration of pulmonary function test devices and HIS was achieved by linking up the spirometer,bluetooth height & weight digital measuring equipment,barcode scanner,back tracking system and the switch to HIS.Results The system contributed to enhancing the flow and efficiency of clinical service.Conclusion The system standardizes the flow and quality control of pulmonary function test,implements patient data sharing between the outpatient department and ward and facilitates scientific research data acquisition,and thus is worthy promoting clinically.

Principles and progress in applications of presbyopia-correcting intraocular lens / 国际眼科杂志(Guoji Yanke Zazhi)

@#As cataract surgery has developed from the simple vision rehabilitation surgery into a more challenging refractive surgery, more and more people wish to improve vision and take off glasses through surgeries. Monofocal intraocular lens can solve the distant vision problem of patients, but near vision patients still need to wear glasses due to the loss of crystal regulation. Hence, presbyopia correction intraocular lens emerged accordingly. In the past decades, we have found and endeavored to solve the problems accompanied in the application of bifocal intraocular lens. On the bifocal basis, some newly-produced and extensively-applied presbyopia correction intraocular lens in the market have become hot products. Presbyopia correction is still a huge challenge for cataract surgery, and this paper will summarize the design principles and clinical application effects of some popular types of presbyopia correction intraocular lens.

Does Cimicifuga racemosa have the effects like estrogen on the sublingual gland in ovariectomized rats?

BACKGROUND: Cimicifuga racemosa is one of the herbs used for the treatment of climacteric syndrome, and it has been cited as an alternative therapy to estrogen. Apart from hectic fevers, dyspareunia and so on, dry mouth also increase significantly after menopause. It has not yet been reported whether C. racemosa has any impact on the sublingual gland, which may relate to dry mouth. In an attempt to determine this, we have compared the effects of estrogen and C. racemosa on the sublingual gland of ovariectomized rats. RESULTS: HE staining showed that the acinar cell area had contracted and that the intercellular spaces were broadened in the OVX (ovariectomized rats) group, while treatment with estradiol (E2) and iCR (isopropanolic extract of C. racemosa) improved these lesions. Transmission electron microscopy showed that rough endoplasmic reticulum expansion in mucous and serous acinar epithelial cells and apoptotic cells was more commonly seen in the OVX group than in the SHAM (sham-operated rats) group. Mitochondria and plasma membrane infolding lesions in the striated ducts were also observed. These lesions were alleviated by both treatments. It is of note that, in the OVX + iCR group, the volume of mitochondria in the striated duct was larger than in other groups. Immunohistochemical staining showed that the ratio of caspase-3 positive cells was significantly increased in the acinar cells of the OVX group compared with the SHAM group (p < 0.05); and the MA (mean absorbance) of caspase-3 in the striated ducts also increased (p < 0.05). Estradiol decreased the ratio of caspase-3 positive cells and the MA of caspase-3 in striated ducts significantly (p < 0.05). ICR also reduced the ratio of caspase-3 positive cells and the MA in the striated ducts (p < 0.05), but the reduction of the MA in striated ducts was inferior to that of the OVX + E2 group (p < 0.05). CONCLUSION: Both estradiol and iCR can inhibit subcellular structural damage, and down-regulate the expression of caspase-3 caused by ovariectomy, but their effects were not identical, suggesting that both drugs confer a protective effect on the sublingual gland of ovariectomized rats, but that the specific location and mechanism of action producing these effects were different.

Active compounds from Baoyuan decoction with myocardial protection activities and their potential targets / 中国中药杂志

Baoyuan decoction (BYD) is a classical Chinese formula for coronary heart disease with Qi deficiency, blood stasis tonifying Qi and Yang deficiency. However, the chemical material basis and underlying action mechanisms of BYD still lack systemic study. In order to clarify the active compounds and the potential action mechanisms of BYD, the oxygen-glucose deprivation/recovery (OGD/R)-induced H9c2 cells injury models was used to screen the monomeric compounds of BYD with myocardial protection activity. PubChem's BioAssay database was then used to analyze the potential targets of active monomeric compounds, classify the predicted biological targets, and analyze the internal relation between active compounds of BYD and biological targets. The screening results showed that BYD aqueous extract and 17 monomeric compounds could significantly increase the survival rate of OGD/R-induced H9c2 myocardial cells. The results of virtual targets screening study showed that 15 monomeric compounds and the potential mechanisms for myocardial protection were related to oxidative stress pathway, calcium ion pathway, mitochondrial protection, anti-apoptosis, etc. These results verified that BYD had myocardial protection effect, which was obtained by network regulation of multi-components and multi-targets. All these results provide the theoretical basis and references for the clinical usage of BYD in treatment of coronary heart disease.

Solid-state fermentation of Acanthogobius hasta processing by-products for the production of antioxidant protein hydrolysates with Aspergillus oryzae

Functional properties and antioxidative activity of a protein hydrolysate prepared from Acanthogobius hasta processing by-product protein during solid-state fermentation with Aspergillus oryzae were investigated. Overall, protease activity increased with the degree of hydrolysis (DH) decreased during solid-state fermentation. All the protein hydrolysate had excellent solubility, possessed interfacial properties, and varying degrees of antioxidant activity which were governed by their concentrations and DH, molecular weight distribution and amino acid composition. After 5 days fermentation, the DH of the protein hydrolysate was 31.23%. The protein hydrolysate had the highest total hydrophobic amino acid content, the highest DPPH scavenging activity, reducing power, and the chelating activity. The radical-scavenging activity of the hydrolysates at 6 mg/mL was 78.6%. The reducing power of protein hydrolysate at the range of 0-6 mg/mL was lower than that of BHA at the range of 0-60 µg/mL, while the chelating activity of APs was similar to that of BHA at the range of 0-60 µg/mL. Moreover, the protein hydrolysate showed good emulsifying and foaming properties over a wide pH range from 2 to 12. Therefore, solid state fermentation provided a suitable and low-cost method for converting Acanthogobius hasta processing by-product protein into antioxidant protein hydrolysates.

Associations of SUMO4 polymorphisms with tacrolimus concentrations in Chinese renal transplant recipients / 药学学报

The study aims to investigate the associations of SUMO4 polymorphisms with tacrolimus concentrations in Chinese renal transplant recipients. Blood samples and clinical data were collected from 132 renal transplant recipients with tacrolimus treatment. CYP3A5*3 genotypes were detected by PCR-RFLP, and SUMO4 (rs237024, rs237025) genotypes were detected by Sequenom® MassARRAY system. SUMO4 rs237024 and rs237025 genotypes were in complete linkage disequilibrium (D' = 1). The dose-adjusted concentration of tacrolimus in SUMO4 rs237024A-rs237025A (GA-GA +AA-AA) carriers was considerably higher than that in GG-GG carriers (P < 0.05). After stratification by CYP3A5*3 genotypes, SUMO4 rs237024A-rs237025A carriers (GA-GA+AA-AA) had a higher dose-adjusted tacrolimus concentration than that in GG carriers in CYP3A5 expresser (P < 0.05). The results illustrated that SUMO4 rs237024 and rs237025 polymorphisms were associated with tacrolimus concentrations, and the test of these genotypes may be useful for individualized medicine of tacrolimus.

Effects of crowding and sex on fecal cortisol levels of captive forest musk deer

BACKGROUND: Restricted space and close contact with conspecifics in captivity may be stressful for musk deer, as they are highly territorial and solitary in the wild. So we tested the effects of crowding on stress of forest musk deer (Moschus berezovskii) in heterosexual groups, using fecal cortisol analysis as a non-invasive method. 32 healthy adults during non-breeding seasons were chose as our experimental objects. Group 1 was defined as higher crowding condition, with 10-15 m²/deer (6 enclosures, 10♀ and 6♂); group 2 was defined as lower crowding condition, with 23-33 m²/deer (6 enclosures, 1010♀ and 6♂). Every enclosure contained 1 male and 3 female. These patterns had been existed for years. RESULTS: The results showed that females in lower crowding condition (217.1 ± 9.5 ug/g) had significantly higher fecal cortisol levels than those in higher crowding condition (177.2 ± 12.1 ug/g). Interestingly, crowding seemed have no effect on male fecal cortisol levels (148.1 ± 9.1 ug/g and 140.5 ± 13.3 ug/g, respectively). At both groups, cortisol was significantly lower in males than in females. CONCLUSIONS: These results showed that chronic crowding may affect stress status of captive forest musk deer. The captive environment should consider the space need for musk deer.