[The combined application of oligoclonal bands in cerebrospinal fluid and IgG intrathecal synthesis indicators and biochemical markers in the diagnosis of multiple sclerosis].

Objective: To establish and verify a diagnostic model for distinguishing multiple sclerosis (MS) from other neurological diseases with similar symptoms by usingcerebrospinal fluid oligoclonal band (CSF-OCB)combined with IgG intrathecal synthesis indicators and biochemical markers. Methods: Multiple sclerosis (MS) patients admitted to the Neurology Department of Beijing Tiantan Hospital affiliated with Capital Medical University from January 2014 to December 2022 were selected as the case group, while patients with similar neurological symptoms were selected as the control group. Using the case-control study design, a retrospective analysis was conducted on the detection of age, gender, oligoclonal bands in cerebrospinal fluid, IgG intrathecal synthesis indicators and biochemical indicators for all study subjects. The differential diagnosis model was determined by the multiple logistic regression analysis, and the receiver operating characteristic (ROC) curve was used to analyze the diagnostic efficiency of the differential diagnosis model for neurological diseases with similar symptoms to MS and other conditions. Results: This study included 167 patients in the case group and 335 patients in the control group, of which 128 patients in the case group and 265 patients in the control group were used to construct the model, and 39 patients in the case group and 70 patients in the control group were used for model validation. The differential diagnostic model constructed by a multivariate logistic regression model was Y=0.871×CSF-OCB-0.051×CSFprotein-0.231×CSFchloride+1.183×gender-0.036×LDH+35.770. The model showed that the area under the curve, sensitivity and specificity were respectively 0.916, 87.3% and 87.6%. The Delong test results showed that the diagnostic efficacy of the model was significantly different from OCB, IgG intrathecal synthesis indicators, and OCB combined with IgG intrathecal synthesis indicators (P<0.05). The new model validation showed that the actual diagnostic consistency rate for the MS group was 84.6%, while the actual diagnostic consistency rate for the control group was 90.0%. Conclusion: This study combines OCB, IgG intrathecal synthesis indicators, and biochemical indicators to establish a diagnostic prediction model for neurological diseases with similar clinical symptoms in MS. This model may have good differential diagnostic value and can better assist clinical diagnosis in the early stages of disease progression in MS patients.

[Comparison of four-coagulation-tests values in normal pregnant women during early and late pregnancy and the influence of age].

Objective: To explore and compare the reference ranges of four coagulation tests in normal pregnant women during early and late pregnancy and the influence of age. Methods: Values of four coagulation tests from 4 974 pregnant women, who gave single birth at Peking University First Hospital, Obstetrics and Gynecology Hospital of Fudan University, West China Second University Hospital, Peking University Third Hospital and Shengjing Hospital of China Medical University from February 2017 to July 2020, were measured and analyzed in this study, including prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fib) and thrombin time (TT). The four normal reference ranges of coagulation during early and late pregnancy phases were expressed as P2.5-P97.5. The difference of two pregnancy phases was compared by non-parametric test of two related samples. And the difference between pregnant women of advanced and non-advanced age in the same pregnancy phase was compared by independent sample non-parametric test. Chi-square test was used to compare the incidence of pregnancy complications in different coagulation reference ranges. Results: The reference ranges of PT of normal pregnant women's early and late pregnancy were 10.0-13.9 s and 9.6-12.3 s, the reference ranges of APTT were 22.6-35.3 s and 22.4-30.9 s, the reference ranges of Fib were 2.4-5.0 g/L and 3.0-5.7 g/L, the reference ranges of TT were 12.0-19.0 s and 11.5-18.4 s. Compared with early pregnancy, PT, APTT and TT shortened significantly, while the Fib significantly increased in late pregnancy (all P<0.001). PT, APTT and TT of advanced and non-advanced age pregnant women were significantly different (all P<0.01). Compared with the ranges of non-pregnant population, more pregnant women were included in the normal pregnant reference ranges of PT in early pregnancy and APTT in the early and late pregnancy, while the incidence of pregnancy complications had no significant differences (all P>0.05). The incidence of fetal distress was higher and the incidence of preterm birth was lower in the reference range of PT in late pregnancy. The incidence of gestational diabetes mellitus was higher in the early and late gestational Fib reference ranges, and the incidence of hypertensive disorders in pregnancy was higher in the late gestational Fib reference range (all P<0.05). Conclusions: The coagulation function of pregnant women increases significantly with the growth of pregnancy, and there is a significant difference between advanced significantly and non-advanced age pregnant women. The recommended ranges of normal pregnant women's early and late pregnancy PT are 10.0-13.9 s and 9.6-12.3 s, the recommended ranges of APTT are 22.6-35.3 s and 22.4-30.9 s, the recommended ranges of TT are 12.0-19.0 s and 11.5-18.4 s. The appropriate ranges of normal pregnant women's early and late pregnancy Fib still need further exploration.

[Effects of 4'-O-methylochnaflavone on endothelial dysfunction induced by palmitic acid in rat cavernous endothelial cells].

OBJECTIVE: To investigate the effect of biflavonoid 4'-O-methylochnaflavone (MF) on palmitic acid-induced endothelial dysfunction in rat cavernous endothelial cells (RCECs). METHODS: The isolated RCECs were commercially available and randomly divided into four groups: normal+BSA group (NC group), palmitic acid (PA) group, MF group, and icariside Ⅱ (ICA Ⅱ) group. The protein expression levels of protein kinase B (PKB/AKT) and endothelial nitric oxide synthase (eNOS) in each group were evaluated via Western blotting. The differences in the intracellular nitric oxide of RCECs treated by MF or ICA Ⅱ were detected by DAF-FM DA that served as a nitric oxide fluorescent probe. Effects of MF and ICA Ⅱ on cell proliferation of PA-stimulated RCECs were determined via CCK-8 assay. RESULTS: The content of nitric oxide in RCECs was significantly increased after the treatment of MF and ICA Ⅱ in comparison with the NC group (P < 0.05). Moreover, compared with ICA Ⅱ group, MF demonstrated a more obvious effect in promoting nitric oxide production (P < 0.05). Compared with the NC group, the expression levels of eNOS and AKT in the PA group were significantly decreased, indicating that a model for simulating the high-fat environment in vitro was successfully constructed (P < 0.05). Meanwhile, the intervention of MF and ICA Ⅱ could effectively increase the expression of eNOS and AKT, suggesting that MF and ICA Ⅱ could promote the recovery of endothelial dysfunction caused by high levels of free fatty acids (P < 0.05). The results of CCK-8 assays showed that PA could significantly reduce the proli-feration ability of RCECs (P < 0.05). Furthermore, the decreased cell viability induced by PA was significantly elevated by treatment with ICA Ⅱ and MF (P < 0.05). CONCLUSION: In RCECs, MF and ICA Ⅱ could effectively increase the content of nitric oxide. The down-regulation of the expression of proteins associated with the AKT/eNOS pathway after PA treatment revealed that this pathway was involved in the development of endothelial dysfunction, which could be effectively reversed by MF and ICA Ⅱ. In addition, the cell proliferation ability was significantly decreased following PA treatment, but MF and ICA Ⅱ could restore the above changes. Overall, biflavonoid MF has an obvious repairing effect on PA-stimulated endothelial dysfunction.

[Clinical and histopathological features of immune checkpoint inhibitor-related myositis in patients with advanced non-small cell lung cancer].

Objective: To investigate the clinicopathologic features and outcome of myositis in patients with advanced non-small cell lung cancer treated with immune checkpoint inhibitors. Methods: The patients diagnosed with immune checkpoint inhibitor-related (ICI) myositis in the database of Respiratory Pathology Center of The First Affiliated Hospital, Guangzhou Medical University from June 2019 to December 2020 were retrospectively analyzed. We reported the muscle histology and main clinical manifestations of the patients in this study. Seven patients with advanced non-small cell lung cancer and ICI related myositis were examined; all of the patients were male, with a median age of 64 (range 42-79) years. Results: All seven patients developed myositis under therapy (three for pembrolizumab, three for sintilimab, and one for camrelizumab). Median delay between ICI initiation and myositis onset was 45 (range 15-176) days. Clinical manifestations were dominated by acute or subacute myalgia and limb weakness. Four patients had evidence of myocarditis. In all of the 7 patients, creatine kinase levels were elevated (median 2 354.4, range 468.6-19 709.2 U/L), while myositis-associated antibodies Ro-52 were positive in four patients. Muscle biopsy showed evident multifocal necrotic myofibers and infiltration of inflammation in two patients. Other patients only showed non-specific endomysial inflammation. Infiltration of inflammation mainly consisted of CD8+ T cells and CD68+ histocytes. After the identification of ICI related myositis, ICI treatment was withdrawn in all patients; 6 patients received corticosteroids therapy. All patients had shown marked clinical improvement. Conclusions: ICI myositis presents with remarkably homogeneous and unique clinicopathologic features, and half of the patients exhibit heightened risk for adverse cardiovascular events, which can be life-threatening if not treated in time. Timely identification of these patients, ICI withdrawal and rapid initiation of corticosteroids therapy can significantly improve patient outcome and/or save patients' lives.

Detection of blaKPC and blaNDM genes by duplex PCR with lateral flow dipsticks from sterile body fluid samples.

Duplex polymerase chain reaction with lateral flow dipsticks (duplex PCR-LFD) was developed for the simultaneous detection of beta-lactamase Klebsiella pneumoniae carbapenemase (blaKPC ) and beta-lactamase New Dehli metallo-beta-lactamase (blaNDM ) genes in body fluid samples. This method was validated using well-characterized isolates. The assessment of the specificity of duplex PCR-LFD showed that there was no cross-reactivity with other targets. The detection limit of the duplex PCR-LFD assay was 20 CFU per ml for blaKPC and blaNDM . Among 177 sterile body fluid samples tested by the duplex PCR-LFD assay, 40 were blaKPC -positive and five were blaNDM -positive. The results obtained from 122 corresponding Gram-negative bacteria which were isolated from these clinical samples and tested by duplex PCR-LFD assay showed that there were 37 strains carrying blaKPC genes in 40 blaKPC -positive samples and three strains carrying blaNDM genes in five blaNDM -positive samples. Statistical analysis indicated that there was no significant difference between the direct detection of blaKPC and blaNDM genes in clinical sterile body fluid samples and their corresponding clinical isolates. Therefore, duplex PCR-LFD can be effective for the simultaneous detection of blaKPC and blaNDM in clinical isolates and directly from clinical samples, which may be helpful for the administration of appropriate antimicrobial treatment.

[Determination of Methoxyacetic acid in urine by pre-column derivatization-liquid-liquid microextraction coupled with gas chromatography].

Objective: To establish a method for determining methoxyacetic acid in urine by pre-column derivatization-liquid-liquid microextraction coupled with gas chromatography (GC) . Methods: Phosphate buffer solution, tert-butoxyacetic acid (internal standard) and pentafluorobenzyl bromide (derivative) were added to the urine sample. After derived in a water bath at 90 ℃ for 40 min, the mixture was cooled and filtered, then the dichloromethane was used as an extractant. After being shaken and centrifuged, the lower organic phase was sucked and injected into a gas chromatograph, separated by a DB-5 capillary column, and detected by an ECD detector. Results: The linear range of the method was 0.6~60.0 mg/L with the correlation coefficients (r) above 0.999. The average recovery was76.6%~110.7%, the inter-day precision was 8.00%~8.82%, and the detection limit was 0.13 mg/L. Conclusion: The method was founded to be high sensitivity, low organic reagent usage and green. So it is suitable for the detection of methoxyacetic acid in urine of occupational exposure to ethylene glycol monomethyl ether.

[Determination of bisphenol S in urine by vortex-assisted liquid-liquid microextraction with high performance liquid chromatography].

Objective: To establish a method for the determination of bisphenol S in urine using dispersive liquid-liquid microextraction (DLLME) coupled with high performance liquid chromatography (HPLC) . Methods: The acetonitrile, octanol were used as extraction solvent, dispersive solvent respectively, for the preconcentration of bisphenol S. The optimal extraction conditions were optimized by single factor rotations, and methodological performance index were tested. Results: The linear correlation coefficient of bisphenol S in the range of 0.0-160 µg/L is greater than 0.999. The detection limit of this method was 0.76 µg/L, and the recovery rates were 88.06%-103.81%. The intra-and inter-day precisions were 1.78%-2.85% and 2.65%-4.25%, respectively. Conclusion: The method is reliable and sensitive. It is suitable for the determination of bisphenol S in urine samples for occupational exposure populations and non-professional.

[Determination of metabolites of styrene in urine by dispersive liquid-liquid microextraction coupled with high performance liquid chromatography].

Objective: To establish a method for the determination of mandelic acid and phenylglyoxylic acid in the urine of styrene by dispersive liquid-liquid microextraction-high coupled with high performance liquid chromatography. Methods: N-octanol was used as an extractant and ethanol was used as a dispersing agent. The phenylglycolic acid and phenylglyoxylic acid in the urine were extracted, and the upper liquid was taken after vortexing and centrifuged, and then was injected into HPLC for analysis. Results: The linear correlation coefficient of the concentration of phenylglycolic acid in the range of 0~10.0 mg/L was greater than 0.999. The detection limit of the method was 9.9 µg/L, the recovery rates were 86.1%~101.6%. The intraday RSDs of the method were 1.07%~3.76%, and the interday RSDs were 1.24%~3.33%. The linear correlation coefficient of phenylglyoxylic acid in the range of 0.0~2.0 mg/L is greater than 0.999. The detection limit of the method was 2.6 µg/L, the recovery rates were 88.8%~100.3%. The intraday RSDs of the method were 1.02%~ 3.17%, and the interday RSDs were 1.59%~2.41%. Conclusion: The method has low detection limit, high enrichment ratio and good sensitivity, and is suitable for determination of phenylglycolic acid and phenylglyoxylic acid in urine of occupational exposure to styrene.

[Determination of manganese in human urine by dispersive ionic liquid-liquid microextraction-graphite furnace atomic absorption spectrometry].

Objective: To establish a method for the determination of manganese in urine with graphite furnace atomic absorption spectrometry (GFAAS) by using ionic liquid microextraction. Methods: The ethanol, 8-hydroxyquinoline and ionic liquid 1-octyl-3-methyl-imidazolium hexafluorophosphate were used as dispersive solvent, chelating agent and extraction solvent respectively, for the preconcentration of manganese. After the optimal extraction conditions were optimized by single factor rotations, evaluate the performance indicators such as methodological precision, accuracy, and detection limit. Results: The linear range of urine manganese was 0.0-1.6 µg/L, and the correlation coefficient of standard curve line was 0.992, the detection limit was 0.03 µg/L, the recovery of sample spiked was 84.90%-96.50%, and the relative standard deviation was 0.36%-1.84%. Conclusion: The method has the advantages of low detection limit, high recovery rate and high sensitivity. It is suitable for the determination of manganese in urine samples from occupational exposure populations and the general population.

[Epidemiological analysis on a family cluster of COVID-19].

Objective: To understand the possible transmission route of a family cluster of COVID-19 in Zhengzhou and the potential infectivity of COVID-19 in incubation period, and provide scientific evidence for the timely control of infectious source and curb the spread of the epidemic. Methods: Epidemiological investigation was conducted for a family cluster of COVID-19 (8 cases) with descriptive epidemiological method, and respiratory tract samples of the cases were collected for the nucleic acid detection of virus by RT-PCR. Results: Two primary cases, which occurred on 31 January and 1 February, 2020, respectively, had a common exposure history in Wuhan. The other six family members had onsets on 30 January, 31 January, 1 February (three cases) and 3 February, 2020. Conclusions: In this family cluster of COVID-19, six family members were infected through common family exposure to the 2 primary cases. Five secondary cases had onsets earlier than or on the same day as the primary cases, indicating that COVID-19 is contagious in incubation period, and the home isolation in the early phase of the epidemic might lead to the risk of family cluster of COVID-19.

[Clinical analysis of 30 cases of Castleman disease with different types of thoracic involvement].

Objective: To improve the clinical understanding of Castleman disease (CD) with different types of thoracic involvement, including their clinical features, radiological and pathological findings, diagnosis and current treatment strategies. Methods: Retrospective analysis of 30 patients diagnosed with CD with thoracic involvement and hospitalized between June 2009 and May 2019 in The First Affiliated Hospital of Guangzhou Medical University was performed. Patients were divided into three groups for subsequent analysis based on the clinical data: CD with bronchiolitis obliterans (BO) , unicentric Castleman disease (UCD) without BO, and multicentric Castleman disease (MCD) without BO. Results: Among the 30 patients, there were 5 (16.7%) patients diagnosed with BO, 18 (60.0%) patients had UCD without BO and 7 (23.3%) patients had MCD without BO. The average age of MCD without BO patients was significantly older than that of BO and UCD without BO patients[ (49.29±5.39) ys vs (27.20±3.76) ys and (37.17±2.87) ys; P=0.005 and 0.034, respectively) ]. Pulmonary symptoms were commonly seen in BO group (100%) and MCD without BO group (71.4%) . while no pulmonary symptoms were seen in UCD without BO group. Key abnormal laboratory findings were erythrocyte sedimentation rate (ESR) increase (40%in BO group and 57.1% in MCD without BO group) and hypoxia (60% in BO group and 28.6% in MCD without BO group) . Other abnormal laboratory findings seen in MCD without BO group included anemia and IgG increase (both 57.1%) . Notably, all patients in BO group had extremely severe mixed ventilation dysfunction in the lung function test. CT scan showed lung parenchyma involvement in BO group (100%) , in UCD without BO group (11.1%) featured by solitary pulmonary nodule and in MCD without BO group (57.1%) featured by diffuse lesions in bilateral lungs. The size of lymph nodes was significantly smaller in MCD without BO group comparing to that in BO group and UCD without BO group[short diameter (1.83±0.51) cm vs (4.73±1.63) cm and (3.62±0.26) cm; P=0.006 and 0.011, respectively]. All patients (100%) in the BO group had a pathological type of transparent vascular variant while the same pathological type accounts for 88.9% in UCD without BO patients. The predominantly pathological type (57.1%) was plasma cell variant in the MCD without BO group. Oral ulcers presented in all patients in BO group but were relieved after the mass resection and immunomodulatory therapy, but the pulmonary symptoms were still progressively aggravated. Thoracoscopic mass excision was the main treatment for UCD without BO patients while chemotherapy, immunomodulatory and targeted therapy were commonly used for MCD without BO treatment. Conclusion: The age, clinical symptom, laboratory finding, lung function, imaging manifestation, pathology, treatment and prognosis were different among the three groups. This classification could improve clinical understanding of the disease.

[Transbronchial lung cryobiopsy in diffuse lung disease: a pathological analysis of 173 cases].

Objective: To evaluate the value of transbronchial lung cryobiopsy (TBCB) in pathological diagnosis for diffuse lung disease. Methods: The clinicopathological data of 173 patients from the first affiliated hospital of Guangzhou medical university between Jaunary 2017 and June 2019 with transbronchial lung cryobiopsy of diffuse lung disease were retrospectively analyzed and summarized with review. Among 173 cases, TBCB and conventional transbronchial lung biopsy (TBLB) were performed in 54 patients. The size of biopsy samples and diagnostic yield were compared. Results: Among 173 cases, the diagnostic yield was 85.54% (148/173) , 160 (92.49%) cases provided definite diagnosis and valuable pathological results, according to age, sex, occupation, past history, contact history, smoking history, laboratory serology and imaging findings. Among 160 cases, there were 72 cases of known etiology (45.00%), 27 cases of idiopathic interstitial pneumonia (16.88%), 7 cases of granulomatous lesions (4.38%) and 54 cases of other types (33.75%). With TBCB and TBLB in 54 patients, the specimens sizes of TBCB and TBLB were (3.3±1.3) mm(2) and (1.0±0.3) mm(2) respectively (t'=12.67 P<0.01) . The diagnostic yields of TBCB and TBLB were 81.48% (44/54) and 42.59% (23/54) respectively (χ(2)=17.33, P<0.01) . The diagnostic yields of TBCB and TBLB for interstitial lung diseases were 48.15% (26/54) and 5.56% (3/54) respectively (χ(2)=24.94, P<0.01) . However, the diagnostic yields of TBCB and TBLB for the other diffuse lung disease except interstitial lung diseases were 33.33% (18/54) and 37.04% (20/54) respectively, with no significant difference (χ(2)=0.1624, P=0.687). Conclusion: Compared with TBLB, TBCB has obvious advantages and application value in the diagnosis of diffuse pulmonary diseases, especially interstitial pulmonary diseases.

Suppressing the secretion of exosomal miR-19b by gw4869 could regulate oxaliplatin sensitivity in colorectal cancer.

Oxaliplatin is commonly used in managing malignancy, including colorectal cancer. While treatment often fails due to decreased drug sensitivity, the mechanisms involved are not clear. In this study, we investigate how exosomal miR-19b participates in oxaliplatin sensitivity and then prove that miR-19b down-regulates oxaliplatin sensitivity of sw480 cells. We found that suppressing the secretion of exosomal miR-19b with gw4869 promotes sw480 cell oxaliplatin sensitivity. Our combined results demonstrate for the first time that miR-19b regulates the oxaliplatin sensitivity of sw480 cells and provides a unique mechanism mediated by gw4869 to modulate oxaliplatin sensitivity by suppressing exosomal miR-19b release.

[Clinical analysis of 3 cases of eosinophilic granulomatosis with polyangiitis].

Objective: To improve the clinical recognition of eosinophilic granulomatosis with polyangiitis(EGPA) in clinical manifestations, diagnosis and treatment. Methods: The clinical manifestations, pathological characteristic, imaging manifestations, diagnosis and the therapy of three patients with EGPA were presented. Results: These 3 patients had asthma-like symptoms and extrapulmonary manifestations of systemic vasculitis. They were 20, 40 and 44 years old. All of them were female.They denied exposure or contact. Chest radiographic examination showed that the most common features were nodule shadow and tree-in-bud in the lung. The pathological manifestation was characterized by hypereosinophilia, high total IgE(over 300 KU/L) and high CRP(over 14.1mg/L). The FeNO of 2 patients was over 100ppb. The ANCA of these 3 patients was negative. The pulmonary pathology was observed had eosinophil infiltration in the alveolar, interstitial and vessel for 3 cases. The clinical manifestations were nonspecific. All patients were treated by glucocorticoid and immune-inhibitor(alkylating agents or purine synthesis inhibitors) therapy. Because patients were complicated with other organs involved, they needed long-time treatment. Conclusions: This disease is diverse and complex, with a lack of pathognomonic symptoms. We should highly suspect eosinophilic granulomatosis with polyangiitis, when the patients present severe asthma and eosinophilia. Early detection, early treatment, and the prognosis could be better.

Carbon monoxide releasing molecule-2 suppresses proliferation, migration, invasion, and promotes apoptosis in non-small cell lung cancer Calu-3 cells.

OBJECTIVE: Non-small cell lung cancer (NSCLC) is the most common type of lung cancer, which is the leading cause of cancer-related morbidity and mortality worldwide. The carbon monoxide-releasing molecules (CO-RMs) are transition metal carbonyls with the capacity to release carbon monoxide (CO). The aims of our study were to assess the effects and underlying mechanisms of CO-releasing molecules-2 (CORM-2) on proliferation, migration, invasion and apoptosis in NSCLC cells, and to evaluate its potential application for lung cancer. MATERIALS AND METHODS: NSCLC cells Calu-3 were treated with CORM-2, negative control and blank control. Cell proliferation, migration and invasion were assessed by cell Counting Kit-8 (CCK-8), scratch assay and matrigel invasion chamber experiment, respectively. Apoptosis was measured by flow cytometry. Real-time PCR and Western blot were applied to examine the expression of apoptosis-related molecules on mRNA and protein levels. RESULTS: CORM-2 markedly attenuated proliferation, migration and invasion of Calu-3 cells. CORM-2 treatment also significantly reduced the ratio of B cell lymphoma 2 (Bcl-2)/B cell lymphoma 2 associated X protein (Bax) while increased expression of caspase-3 and cytochrome c. The optimal dose of CORM-2 for Calu-3 cells was 100 µM. CONCLUSIONS: CORM-2 modulates biological functions of NSCLC cells and may provide a novel therapeutic strategy for lung cancer.

[Significance of the combined detection of routine blood test, serum iron and hemoglobin electrophoresis in screening thalassemia in non- high incidence area].

Objective: To evaluate the role of combined detections of routine blood test, serum iron and hemoglobin electrophoresis in screening thalassemia in non- high incidence area. Methods: Peripheral blood and serum samples of 1 000 outpatients from the department of hematology and the department of gynecology and obstetrics were obtained. Common mutations of thalassemia were detected by using GAP- PCR and reverse dot blotting, and Sanger sequencing was performed to discover rare mutations of α- and ß- thalassemia. Routine blood test, serum iron and hemoglobin electrophoresis were also performed for every patient. Results: Among 1 000 samples, 225(22.5%)are detected as α-thalassemia, 403(40.3%)ß-thalassemia and 15(1.5%)composite thalassemia. Among 225 α-thalassemia patients, 28 were silent, 138 were intermedia, and 59 were HbH disease. Of 403 ß-thalassemia, 390 were carriers, 7 were double heterozygote, and 6 were homozygote. In all samples, there were 357 patients detected with no common mutations, 38 patients had higher result values for both MCV and MCH and none detected with thalassemia gene. There were 48 patients who had higher serum iron but normal or lower MCV, 42 of them(87.5%)had thalassemia gene. Furthermore, 38 patients showed abnormal hemoglobin electrophoresis, 35 of them were HbH disease, while the other 3 were HbF- related thalassemia. Five patients showed abnormal hemoglobin electrophoresis, lower MCV and MCH, as well as higher serum iron, had no frequent mutation but rare ones. Conclusion: Patients with higher MCV and MCH can mostly be excluded to have thalassemia, while higher serum iron represents thalassemia possibility and can provide a preliminary indication of thalassemia type, and last but not least abnormal hemoglobin electrophoresis indicates the disease. It is recommended to further carry out sequencing of rare mutations for those who had abnormal results in the combined screening, and detected with no frequent mutation. Combination of these three examinations can improve the detection efficiency of patients with thalassemia.

[Pulmonary surfactant protein adenosine triphosphate-binding-cassette-A3 gene composite mutations in infant congenital interstitial lung disease: report of a case and review of literature].

Objective: To report a case of the pulmonary surfactant protein(SP) adenosine triphosphate-binding-cassette-A3 (ABCA3) gene mutations in infant congenital interstitial lung disease(ILD), and review the related literature, to investigate the relationships of ABCA3 gene mutation associated with ILD in infants. Method: A 6-months-old boy was hospitalized in the department of Pediatrics of the First Affiliated Hospital of Guangzhou Medical University. The clinical, radiological, histological information from transbronchial lung biopsy (TBLB) and genetic testing in this case was analyzed; 12 reports retrieved on literature search at Pubmed, OVID databases from 2004 to 2015 by using the ABCA3 as keyword were reviewed and analyzed. Result: (1)The patient, a 6-months-old boy, had progressive tachypnea and dyspnea since 4 months old. Physical examination on admission revealed respiratory rate of 78 times/min , heart rate of 187 times/min, SpO2 0.93(mask oxygen-inspiration with 6 L/min), scattered fine moist crackles could be heard over the both lungs, clubbing fingers were found. High-resolution computed tomography(HRCT) revealed diffuse ground-glass opacity, interlobular and intralobular septal thickening. Lung biopsies showed evidences of the alveolar cavity atelectatic changes and interstitial fibrosis. SP-A and SP-B were negative in immunohistochemical stainting. SP-related gene sequence analysis found that there was compound heterozygous missense mutation of ABCA3 gene in c. 1942A>G, c.2701-33G>C and c. 991-105C>A. (2)The review of related literature found that totally 12 cases were reported. The main manifestations were progressive tachypnea and dyspnea, age of onset was between birth and 4 years of age. The imaging characteristics of chest HRCT revealed diffuse infiltration or diffuse ground-glass pattern in the lung. PROGNOSIS: 6 cases died, and 6 cases survived, including 4 cases with pulmonary function disturbance to different degrees; 12 cases had ABCA3 gene mutations, 9 cases had composite ABCA3 gene mutations, in 11 cases the mutation occured in the exon of coding region, in 1 case in the intron, 9 cases had heterozygous mutations, 3 cases had homozygous mutations. Conclusion: The main phenotypes of ABCA3 mutation associated with ILD were full term neonatal respiratory distress syndrome or progressive tachypnea or dyspnea unexplained in infants. The chest HRCT showed two diffuse pulmonary interstitial changes. ABCA3 mutation mainly was multi-site composite mutations and heterozygous mutations in the exon of coding region.