BAG3-Related Myofibrillar Myopathy Presenting as Hypercapnia: A Case Report and Literature Review.

Objective BAG3-related myopathy is a rare condition so far reported in twenty patients worldwide. The purpose of this study was to draw attention to this rare disease and to the fact that BAG3-related myopathy should be considered as a rare differential diagnosis of hypercapnia. Methods We report a sporadic case of a 14-year-old Chinese girl with a de novo p.Pro209Leu mutation in BAG3 and reviewed the literatures for reported cases related to this mutation. Results We described a 14-year-old Chinese girl who presented with gradually appearing symptoms of hypercapnia that required assisted ventilation. The muscle biopsy and the blood whole-exome sequencing results confirmed the diagnosis of myofibrillar myopathy with a de novo p.Pro209Leu mutation in BAG3. Totally twenty-one patients from twenty families with a confirmed diagnosis of BAG3-related myopathy were reported to date, including this patient and literature review. The male to female ratio was 11:10 and most showed initial symptoms in the first decade of life. Most patients presented toe/clumsy walking or running as the onset symptom, followed by muscle weakness or atrophy. Creatine kinase levels were elevated in fourteen patients and were normal in three. Eighteen patients developed respiratory insufficiency during the disease course and thirteen (one could not tolerate non-invasive assisted ventilation) required non-invasive assisted ventilation for treatment. Except for one not reported, heart involvement was found in seventeen patients during the disease course and seven underwent heart transplantation. Z-disk streaming and aggregation could be observed in most of the patients' muscle histology. In the long-term follow-up, five patients died of cardiac or respiratory failure. Conclusion BAG3-associated myopathy is a rare type of myofibrillar myopathy. It should be considered as a rare differential diagnosis of hypercapnia.

[Clinical Features of Coccidioidomycosis:Analysis of 33 Chinese Cases].

Objective To summarize the characteristics of Chinese coccidioidomycosis cases, improve the diagnosis and treatment of this disease and prevent misdiagnosis as well as therapeutic error.Methods Search in databases including Medline,Wanfang,and CNKI using "Coccidioidomycosis" and "China" as index words yielded 23 articles that reported a total of 32 Chinese coccidioidomycosis cases.In addition,one patient with disseminated coccidioidomycos was treated in our center in April 2016.The demographic data,site of infection,clinical manifestations,past medical history,exposure history,imaging and laboratory findings,and pathological features of these 33 patients were analyzed.Results Among these 33 patients,7(21.2%)had visited an epidemic area and 6(18.2%)were immunocompromised.The disease involved the respiratory system,skin,bone,central nervous system,cornea,and stomach in 24,6,3,2,1,and 1 patients,respectively.Eight patients (24.2%) had multiple system involvement,and three of them died.The imaging findings included pulmonary nodules(n=14),mediastinal lymphadenopathy(n=5),solid shadow(n=4),cavity(n=4),pleural effusion(n=3),multiple plaques(n=2)and masses(n=2).Coccidiolys cysts were detected in the affected tissues(n=28)or in pus,exudate or pleural smear(n=3);in addition,coccidioides mycelium and spores were found in the sputum,pus,and tissue cultures in 4 cases,among whom only 2 cases were confirmed by serological examination.The treatments included triazoles(n=20),systemic or local administration of amphotericin B(n=13),surgical resection of the lesion(n=8),and intravenous gamma globulin(n=1).Five patients died,among whom three had underlying diseases that caused immunosuppression and one was an infant.The prognoses were relatively good in the remaining patients.Conclusions Early diagnosis and proper treatment can achieve good prognosis in coccidioidomycosis patients.Multi-system involvement and immunosuppression are risk factors for poor prognosis of coccidioidomycosis.For these patients,adequate and full-course medication may prevent rapid disease progression.

Improvement of anti-nutritional effect resulting from ß-glucanase specific expression in the parotid gland of transgenic pigs.

ß-Glucan is the predominant anti-nutritional factors in monogastric animal feed. Although ß-glucanase supplementation in diet can help to eliminate the adverse effects, enzyme stability is substantially modified during the feed manufacturing process. To determine whether the expression of endogenous ß-glucanase gene (GLU) in vivo can improve digestibility of dietary ß-glucan and absorption of nutrients, we successfully produced transgenic pigs via nuclear transfer which express the GLU from Paenibacillus polymyxa CP7 in the parotid gland. In three live transgenic founders, ß-glucanase activities in the saliva were 3.2, 0.07 and 0.03 U/mL, respectively, and interestingly the enzyme activities increased in the pigs from 178 days old to 789 days old. From the feed the amount of gross energy, crude protein and crude fat absorbed by the transgenic pigs was significantly higher than the non-transgenic pigs. Meanwhile the moisture content of the feces was significantly reduced in transgenic pigs compared with the non-transgenic pigs. Furthermore, in all positive G1 pigs, ß-glucanase activity was detectable and the highest enzyme activity reached 3.5 U/mL in saliva. Also, crude protein digestion was significantly higher in G1 transgenic pigs than in control pigs. Taken together, our data showed that the transgenic ß-glucanase exerted its biological catalytic function in vivo in the saliva, and the improved performance of the transgenic pigs could be accurately passed on to the offspring, indicating a promising alternative approach to improving nutrient availability was established to improve utilization of livestock feed through transgenic animals.

Chronic Fluoxetine Treatment Upregulates the Activity of the ERK1/2-NF-κB Signaling Pathway in the Hippocampus and Prefrontal Cortex of Rats Exposed to Forced-Swimming Stress.

OBJECTIVES: The aim of this study was to explore whether or not the antidepressant actions of fluoxetine (FLX) are correlated with extracellular signal-regulated kinase 1 and 2 (ERK1/2) and nuclear factor κ-light chain enhancer of activated B cells (NF-κB) in the hippocampus (HC) and prefrontal cortex (PFC) of rats. MATERIALS AND METHODS: A total of 108 male Sprague-Dawley rats were randomly divided into 6 groups of 18 rats each. Group 1 was the control group, while group 2 comprised the depressed model in which rats were subjected to 28 days of forced-swimming stress (FST); groups 3-6 were also subjected to 28 days of FST and treated with FLX once a day for 1 day (group 3; F1d), 1 week (group 4; F1w), 2 weeks (group 5; F2w), or 4 weeks (group 6; F4w). The control group was not subjected to FST or treated with FLX. Behavior tests that included the Morris water maze (MWM) and saccharin preference were performed, and ERK1/2 and NF-κB proteins were assayed using Western blot. RESULTS: The rats in the control group and in groups 5 and 6 (F2w and F4w, respectively) had a significantly shorter average escape latency, needed more attempts in order to successfully cross the platform, and had a greater saccharin preference than those in the depressed group (p < 0.05). In the depressed group, the phosphorylated ERK1/2 (p-ERK1/2) and phosphorylated NF-κB (p-NF-κB) expression in the HC and PFC were lower than in the control group (p < 0.05). Treatment with FLX reversed the changes in the expression of p-ERK1/2 and p-NF-κB in rats in the F2w and F4w groups. CONCLUSIONS: In this study, FLX treatment for 2 weeks or longer reversed the impaired spatial learning, memory, and anhedonia observed in the depressed model rats and upregulated the activities of the ERK1/2-NF-κB signaling pathway.

Malat1 as an evolutionarily conserved lncRNA, plays a positive role in regulating proliferation and maintaining undifferentiated status of early-stage hematopoietic cells.

BACKGROUND: The metastasis-associated lung adenocarcinoma transcription 1 (Malat1) is a highly conserved long non-coding RNA (lncRNA) gene. Previous studies showed that Malat1 is abundantly expressed in many tissues and involves in promoting tumor growth and metastasis by modulating gene expression and target protein activities. However, little is known about the biological function and regulation mechanism of Malat1 in normal cell proliferation. RESULTS: In this study we conformed that Malat1 is highly conserved across vast evolutionary distances amongst 20 species of mammals in terms of sequence, and found that mouse Malat1 expresses in tissues of liver, kidney, lung, heart, testis, spleen and brain, but not in skeletal muscle. After treating erythroid myeloid lymphoid (EML) cells with All-trans Retinoic Acid (ATRA), we investigated the expression and regulation of Malat1 during hematopoietic differentiation, the results showed that ATRA significantly down regulates Malat1 expression during the differentiation of EML cells. Mouse LRH (Lin-Rhodamine(low) Hoechst(low)) cells that represent the early-stage progenitor cells show a high level of Malat1 expression, while LRB (Lin - Hoechst(Low) Rhodamine(Bright)) cells that represent the late-stage progenitor cells had no detectable expression of Malat1. Knockdown experiment showed that depletion of Malat1 inhibits the EML cell proliferation. Along with the down regulation of Malat1, the tumor suppressor gene p53 was up regulated during the differentiation. Interestingly, we found two p53 binding motifs with help of bioinformatic tools, and the following chromatin immunoprecipitation (ChIP) test conformed that p53 acts as a transcription repressor that binds to Malat1's promoter. Furthermore, we testified that p53 over expression in EML cells causes down regulation of Malat1. CONCLUSIONS: In summary, this study indicates Malat1 plays a critical role in maintaining the proliferation potential of early-stage hematopoietic cells. In addition to its biological function, the study also uncovers the regulation pattern of Malat1 expression mediated by p53 in hematopoietic differentiation. Our research shed a light on exploring the Malat1 biological role including therapeutic significance to inhibit the proliferation potential of malignant cells.

Sequence and phylogenetic analysis of surface protein genes of emerging H9N2 influenza viruses isolated from poultry in two geographical regions of China.

Subtype H9N2 avian influenza viruses (AIVs) circulating in China have aroused increasing concerns for their impact on poultry and risk to public health. The present study was an attempt to elucidate the phylogenetic relationship of H9N2 AIVs in two geographically distinct regions of China where vaccination is routinely practiced. A total of 18 emerging H9N2 isolates were identified and genetically characterized. Phylogenetic analysis of hemagglutinin (HA) and neuraminidase (NA) genes confirmed that the isolates belonged to the Y280 lineage. Based on the HA genes, the isolates were subdivided into two subgroups. The viruses from Zhejiang Province were clustered together in Group I, while the isolates from Guangdong Province were clustered together in Group II. Antigenic characterization showed that the tested viruses were antigenically different when compared to the current used vaccine strain. It was notable that 14 out of total 18 isolates had an amino acid exchange (Q→L) at position 216 (226 by H3 Numbering) in the receptor-binding site, which indicated that the virus had potential affinity of binding to human like receptor. These results suggest that the emerging viruses have potential risk to public health than previously thought. Therefore, continuous surveillance studies of H9N2 influenza virus are very important to the prognosis and control of future influenza pandemics.

ß-Glucanase specific expression in the parotid gland of transgenic mice.

The feasibility of using the pig parotid secretory protein promoter to drive the ß-glucanase transgene expression in mouse parotid glands was examined in this study. The parotid gland-specific vector expressing ß-glucanase gene (GLU, from Paenibacillus polymyxa CP7) was constructed. Transgenic mice were produced by the pronuclear microinjection. Both PCR and Southern blot analysis showed that the mice carried the ß-glucanase gene and the ß-glucanase gene could be stably inherited. Furthermore, RT-PCR and northern blot analysis indicated that it was specifically expressed in the parotid. The ß-glucanase activity in the saliva was found to be 0.18 U/mL. After feeding a diet containing 2 % ß-glucan, the average daily gain of transgenic was significantly higher than non-transgenic mice. The crude protein and crude fat concentration in faeces of transgenic mice were significantly reduced compared with that of the non-transgenic mice. These results suggest that the successful expression of foreign ß-glucanase in the animal parotid would offer a promising biological approach to reduce the anti-nutritional effect of ß-glucans in feed.

A strategy for precise and large scale identification of core fucosylated glycoproteins.

Core fucosylation (CF) patterns of some glycoproteins are more sensitive and specific than evaluation of their total respective protein levels for diagnosis of many diseases, such as cancers. Global profiling and quantitative characterization of CF glycoproteins may reveal potent biomarkers for clinical applications. However, current techniques are unable to reveal CF glycoproteins precisely on a large scale. Here we developed a robust strategy that integrates molecular weight cutoff, neutral loss-dependent MS(3), database-independent candidate spectrum filtering, and optimization to effectively identify CF glycoproteins. The rationale for spectrum treatment was innovatively based on computation of the mass distribution in spectra of CF glycopeptides. The efficacy of this strategy was demonstrated by implementation for plasma from healthy subjects and subjects with hepatocellular carcinoma. Over 100 CF glycoproteins and CF sites were identified, and over 10,000 mass spectra of CF glycopeptide were found. The scale of identification results indicates great progress for finding biomarkers with a particular and attractive prospect, and the candidate spectra will be a useful resource for the improvement of database searching methods for glycopeptides.

Comparison of three methods of protein extraction from Dermatophagoides pteronyssinus for two-dimensional electrophoresis.

OBJECTIVE: To explore an effective method of Dermatophagoides pteronyssinus protein extraction suitable for two-dimensional electrophoresis (2-DE) analysis. METHODS: The extracts of Dermatophagoides pteronyssinus were prepared with Coca's solution, lysis buffer of 2-DE, and Trizol reagent, respectively. Bicinchoninic acid (BCA) assay was used to determine the total protein concentration of the samples. The efficiency of different protein extraction methods were evaluated with 2-DE analysis. RESULTS: The concentrations of extracted protein by methods of Coca's solution, lysis buffer, and Trizol reagent were 0.63 g/L, 0.90 g/L, and 0.80 g/L, respectively. The 2-DE analysis results showed that some protein spots in low molecular weight (LMW) range could be detected with the Coca's solution method. With the lysis buffer of 2-DE method, more protein spots in LMW range could be detected, while the medium molecular weight (MMW) protein spots were absent. Several MMW protein spots (174-178 kD and 133 kD) and more LMW protein spots were detected with Trizol reagent method. CONCLUSIONS: Among Coca's solution, lysis buffer of 2-DE, and Trizol reagent, the concentration of extracted protein of Dermatophagoides pteronyssinus by lysis buffer of 2-DE is the highest. However, most protein components of Dermatophagoides pteronyssinus purified mite bodies can be extracted by Trizol reagent, which may generally reflect the whole profile of Dermatophagoides pteronyssinus allergens.

[Primary pulmonary lymphoma: analysis of 18 cases].

OBJECTIVE: To study the clinical characteristics, pathology, diagnosis and treatment of primary pulmonary lymphoma. METHODS: Eighteen cases of primary pulmonary lymphoma diagnosed from Jan 1989 to Feb 2007 were retrospectively analyzed. RESULTS: There were 6 males and 12 females, with a median age of 47.5 years (17-71 years). Fifteen cases were diagnosed by surgical lung biopsy; 1 by percutaneous needle lung biopsy (1/6), 1 by percutaneous needle lung biopsy and bronchoscopic examination at the same time, the other 1 by bronchoscopic examination (1/10). Histological diagnosis showed that 2 cases were Hodgkin lymphoma, 9 mucosa-associated lymphoid tissue lymphoma, 1 follicular lymphoma, 2 diffuse large B cell lymphoma 2 anaplastic large cell lymphoma, 2 non-Hodgkin lymphoma whichcould not be classified because the slides were from other hospitals. The most common symptoms were cough (9/18) and fever (6/18). ESR elevation was common (10/12). CT features included solitary or multiple nodules (14/18), patchy opacities (11/18), consolidations (5/18), pleural effusions (5/18), atelectasis (5/18), and cavities (1/18). Misdiagnosis was found in 11 patients. Treatment modalities included surgical resection, radiotherapy and chemotherapy. Median follow-up time was 11 months (10 d to 205 mon). Thirteen patients were still alive, 4 patients were lost, and 1 patient died. The prognosis was associated with the level of [25.1 x 10(9)/L(18.1 - 39. -1) x 10(9)/L in poor prognosis group, 6.7 x 10(9)/L (5.48 - 8.41) x 10(9)/L in good prognosis group, u = 0.000, P <0.05] leukocytosis (3/3 vs 1/10, P <0.05). CONCLUSIONS: The clinical manifestations of primary pulmonary lymphoma are nonspecific. Misdiagnosis is common. Surgical lung biopsy is necessary for early diagnosis.

[Radio-pathological manifestations of pulmonary sarcoidosis].

OBJECTIVE: To investigate the radio-pathological features of pulmonary sarcoidosis. METHOD: Forty six consecutive patients from January 2000 to August 2005 in Peking Union Medical College Hospital with pathologic features of epithelioid cell granuloma were enrolled in the study. RESULTS: All the cases were confirmed by pathological findings consistent with sarciodosis. Bronchoalveolar lavage (BAL) fluid analysis showed that, the differential count of lymphocytes was 0.47 +/- 0.18, and the CD(4)/CD(8) ratio was 6.63 +/- 4.51. Serum angiotensin converting enzyme level was (47 +/- 16) U/L. Chest computed tomography scan showed bilateral hilar adenopathy (33%, 12/46), well circumscribed pulmonary nodules (35%, 16/46) distributed randomly or along the bronchovascular bundle, patchy areas of alveolar consolidation (28%, 13/46), and bilateral ground glass infiltrations (11%, 5/46). Open lung biopsy, video-assistant thoracoscopic biopsy or mediastinoscopic biopsy were performed in 13 patients, and percutaneous lung biopsy or transbronchial lung biopsy in 33 patients. The characteristic pathologic finding was noncaseating epithelioid cell granulomas, which were embedded in the substance of hyalinization. The granulomas were distributed around blood vessels, lymphatics, or in the bronchial submucosa. Granulomatous vasculitis was noted in some cases. CONCLUSION: The diagnosis of sarcoidosis can be proposed by clinico-radiologic and BAL fluid findings. However, histologic evidence of noncaseating epithelioid cell granulomas and the therapeutic efficacy of glucocorticoids are essential to the final diagnosis.

Development and Preliminary Application of a Peptide Mass Fingerprinting Technique in Proteome Research.

Proteome means the total proteins expressed by the genome in a cell or tissue. Two-dimensional electrophoresis (2-DE) is now the most powerful separating technique and the key separation method used in proteome. Peptide mass fingerprinting (PMF) is becoming a widely used and vastly developed technique for protein identification in 2-D gels. In this research, a systematic method to identify the proteins in polyacrylamide gels by PMF was developed. Proteins were digested in-gel by enzyme and the masses of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The data obtained from PMF were used in protein database search and the protein identification. The proteins from human lung cancer cells isolated by 2-DE were subjected to identification by the PMF method developed in this work. Three spots of proteins in gel were identified as G3P2_HUMAN, UBL1_ HUMAN and TPIS_HUMAN.

[Analysis of protein phosphorylation by combination of IMAC, phosphatase with biological mass spectrometry].

Protein phosphorylation is the most important reversible post-translational modification in cells. Analysis of phosphorylated proteins and identification of their phosphorylation sites is helpful for understanding their biological functions. MALDI-TOF-MS and ESI-Q-TOF-MS play important roles in protein phosphorylation analysis. In this work, immobilized metal affinity chromatography (IMAC) was used to selectively enrich phosphopeptides from protein digest mixtures, and treatment of phosphopeptides with alkaline phosphatase was used to confirm the phosphorylation. Finally, the phosphorylation sites were determined by tandem mass spectrometry analysis and database searching.

[Role of single-breath carbon monoxide-diffusing capacity in monitoring the bleomycin-induced lung toxicity in human].

OBJECTIVE: To investigate if the carbon monoxide-diffusing capacity (D(LCO)) could be the early indictor monitoring the bleomycin-induced lung toxicity (BILT) with retrospection of the influence of bleomycin cumulative dose on the pulmonary functions. METHODS: During June 1985 to October 2000, 42 patients with malignant tumor of ovarian germ cells received chemotherapy containing bleomycin in the department of Obstetrics and Gynecology of Peking Union Medical College Hospital. Twenty three patients (54.76%) among them had >or= 2 courses of chemotherapy with bleomycin and performed >or= 2 measurements of lung function. These 23 patients were included in the study. The forced expiratory volume in one second (FEV(1)), forced vital capacity (FVC), total lung capacity (TLC) and D(LCO) were measured and recorded as % of predicted. The D(LCO) was corrected by hemoglobin concentration [D(LCO) corrected = D(LCO) measured x (9.38 + Hb) divided by (1.76 x Hb)]. All the data were divided into 4 groups according to the cumulative dose of bleomycin (pre-treatment group, < 100 mg group, 101 - 200 mg group and > 201 mg group). The relationship between D(LCO) and bleomycin cumulative dose was examined by linear regression analysis and t-test. RESULTS: The values of FEV(1%)predicted in the 4 groups were 99.83 +/- 16.41 (14), 101.43 +/- 12.32 (42), 99.41 +/- 10.22 (23), and 90.96 +/- 13.63 (12), respectively. The FVC%predicted were 97.74 +/- 18.23 (14), 101.11 +/- 13.95 (42), 96.49 +/- 12.04 (23) and 89.63 +/- 18.20 (12), respectively. The TLC%predicted were 101.22 +/- 10.68 (13), 106.14 +/- 12.16 (40), 102.13 +/- 11.33 (23) and 95.05 +/- 14.06 (11), respectively. There were no statistical significant differences in the parameters among the 4 groups. The D(LCO%)predicted of the 4 groups were 93.27 +/- 12.75 (14), 94.51 +/- 12.50 (40), 80.93 +/- 10.05 (24) and 70.99 +/- 11.69 (15), respectively, and the D(LCO) of the last group (> 201 mg group) was significantly decreased as compared to that of the other groups (P < 0.05). A bleomycin dose-related fall in D(LCO) was observed, y = 100.59 - 0.11x, r = -0.649, P < 0.001. CONCLUSION: The D(LCO) might be the most sensitive indicator of subclinical BILT. However, the cumulative dose of bleomycin did not show significant influence on the FEV(1), FVC and TLC.

Expert consensus on acute exacerbation of chronic obstructive pulmonary disease in the People's Republic of China.

Chronic obstructive pulmonary disease (COPD) is a common disease that severely threatens human health. Acute exacerbation of COPD (AECOPD) is a major cause of disease progression and death, and causes huge medical expenditures. This consensus statement represents a description of clinical features of AECOPD in the People's Republic of China and a set of recommendations. It is intended to provide clinical guidelines for community physicians, pulmonologists and other health care providers for the prevention, diagnosis, and treatment of AECOPD.

[Comparison of the analgesic effect of acupuncture between otopoint-penetrative needling and otopoint-straight needling for cervical type and nerve-root type cervicospondylopathy].

OBJECTIVE: To confirm the better analgesic effect of otopoint-penetrative needling for cervical type and nerve-root type cervicospondylopathy. METHODS: A total of 98 cervicospondylopathy outpatients (50 cases of cervical type and 48 cases of nerve-root type) were randomly divided into treatment group (otopoint-penetrative needling) and control group (otopoint-straight needling) in the light of paring method of comprehensive factors of sexes, ages and the state of disease. The main oto-points used were bilateral Jingzhui Area (AH 13) in combination with Jian-Jianguanjie-Suogu (Shoulder-Shoulder-joint-Collarbone) Area, etc. The simplified McGill Pain Scaling was used to give the score of patient's pain before the treatment, 5 min and 30 min after the treatment. RESULTS: Results of sequential trial indicated that the analgesic effect of otopoint-penetrative needling was significantly superior to that of otopoint-straight needling 30 min after the treatment (P < 0.05). Findings of matched-pair t test showed that no marked differences were found between two groups in the pain scores before the treatment, while after the treatment, the pain scores of otopoint-penetrative needling group were significantly lower than those of otopoint-straight needling group (P < 0.001, 0.01), meaning that the analgesic effect of otopoint-penetrative needling was significantly better than that of otopoint-straight needling 5 min and 30 min after the treatment in both men and women, in both cervical type and nerve-root type patients, and in both young and older patients. CONCLUSION: The analgesic effect of otopoint-penetrative needling is obviously superior to that of otopoint-straight needling.