Endo-beta-N-acetylglucosaminidase (ENGase) is widely distributed in various organisms. The first reported ENGase activity was detected in Diplococcus pneumoniae in 1971. The protein (Endo D) was purified and its peptide sequence was determined in 1974. Three ENGases (Endo F1-F3) were discovered in Flavobacterium meningosepticum from 1982 to 1993. After that, the activity was detected from different species of bacteria, yeast, fungal, plant, mice, human, etc. Multiple ENGases were detected in some species, such as Arabidopsis thaliana and Trichoderma atroviride. The first preliminary crystallographic analysis of ENGase was conducted in 1994. But to date, only a few ENGases structures have been obtained, and the structure of human ENGase is still missing. The currently identified ENGases were distributed in the GH18 or GH85 families in Carbohydrate-Active enZyme (CAZy) database. GH18 ENGase only has hydrolytic activity, but GH85 ENGase has both hydrolytic and transglycosylation activity. Although ENGases of the two families have similar (β/α)8-TIM barrel structures, the active sites are slightly different. ENGase is an effective tool for glycan detection andglycan editing. Biochemically, ENGase can specifically hydrolyze β‑1,4 glycosidic bond between the twoN-acetylglucosamines (GlcNAc) on core pentasaccharide presented on glycopeptides and/or glycoproteins. Different ENGases may have different substrate specificity. The hydrolysis products are oligosaccharide chains and a GlcNAc or glycopeptides or glycoproteins with a GlcNAc. Conditionally, it can use the two products to produce a new glycopeptides or glycoprotein. Although ENGase is a common presentation in cell, its biological function remains unclear. Accumulated evidences demonstrated that ENGase is a none essential gene for living and a key regulator for differentiation. No ENGase gene was detected in the genomes of Saccharomyces cerevisiae and three other yeast species. Its expression was extremely low in lung. As glycoproteins are not produced by prokaryotic cells, a role for nutrition and/or microbial-host interaction was predicted for bacterium produced enzymes. In the embryonic lethality phenotype of the Ngly1-deficient mice can be partially rescued by Engase knockout, suggesting down regulation of Engase might be a solution for stress induced adaptation. Potential impacts of ENGase regulation on health and disease were presented. Rabeprazole, a drug used for stomach pain as a proton inhibitor, was identified as an inhibitor for ENGase. ENGases have been applied in vitro to produce antibodies with a designated glycan. The two step reactions were achieved by a pair of ENGase dominated for hydrolysis of substrate glycoprotein and synthesis of new glycoprotein with a free glycan of designed structure, respectively. In addition, ENGase was also been used in cell surface glycan editing. New application scenarios and new detection methods for glycobiological engineering are quickly opened up by the two functions of ENGase, especially in antibody remodeling and antibody drug conjugates. The discovery, distribution, structure property, enzymatic characteristics and recent researches in topical model organisms of ENGase were reviewed in this paper. Possible biological functions and mechanisms of ENGase, including differentiation, digestion of glycoproteins for nutrition and stress responding were hypothesised. In addition, the role of ENGase in glycan editing and synthetic biology was discussed. We hope this paper may provide insights for ENGase research and lay a solid foundation for applied and translational glycomics.

To revise GBZ 188 Technical Specification for Occupational Health Surveillance based on national laws, regulations, standards, specifications and legal documents of occupational disease, and combination with the actual situation in China. The main modifications are as follows: the occupational health surveillance for workers exposed to toluene (xylene may implement by reference), bromopropane, methyl iodide, ethylene oxide, chloroacetic acid, indium and its compounds, coal tar, coal tarasphalt, asphalt, β-naphthylamine, dust of metal and its compounds(tin, iron, antimony, barium and its compounds), hard metal dust, erionite dust, low temperature, laser, tick-borne encephalitis virus, Borrelia burgdorferi, and human immunodeficiency virus, for scraper or grind operators, and underground workers using squatting or kneeling position, crawling position, side-lying position, or shoulder position for a long period of time are included. The emergency health screening for workers exposed to arsenic, fluorine and its inorganic compounds, and acrylamide are included. The occupational medical examination (OME) for workers exposed to amino and nitro compounds of benzene, phosgene, monomethylamine, organic fluorine and dimethyl sulfate has been adjusted and made mandatory, with corresponding assessments required upon leaving the job. The special occupational health surveillance for workers exposed to mycobacterium tuberculosis and hepatitis virus is removed. The OME conclusion of reexamination is removed, and standardize recheck/additional inspection requirements. The optional items in OME performed before, during and after leaving post are removed, but the optional items in emergency medical examination are retained. Additional OME items are added. The Guideline for OME Summary Reports is added as informative appendix, and so on. The revised GBZ 188 Technical Specification for Occupational Health Surveillance is more scientific and practical.

AIM: To explore the therapeutic effects of Morinda officinalis capsules (MOP) on osteoporosis in ovariectomized rats. METHODS: Six-month-old female Sprague-Dawley rats were induced for postmenopausal osteoporosis (PMOP) by bilateral ovariectomy and divided into seven groups as follows: sham-operated group, ovariectomized (OVX) control group, OVX treated with xianlinggubao (XLGB) (270 mg·kg⁻¹·d⁻¹), OVX treated with alendronate sodium (ALN) (3 mg·kg⁻¹·d⁻¹), and OVX treated with Morinda officinalis capsule (MOP) of graded doses (90, 270 and 810 mg·kg⁻¹·d⁻¹) groups. Oral treatments were administered daily on the 4(th) week after ovariectomy and lasted for 12 weeks. The bone mineral density was evaluated by dual-energy X-ray absorptiometry. The tartrate-resistant acid phosphatase (TRAP), alkaline phosphatase (AKP), and osteocalcin (OC) levels in the serum and plasma were determined by standard colorimetric and enzyme immunoassays methods. Bone biomechanical properties and morphological parameters were analyzed by three-point bending test and histomorphometry respectively. RESULTS: Morinda officinalis capsules at all doses were able to significantly prevent the OVX-induced loss of bone mass due to diminishing serum AKP and TRAP levels while elevating OC level in the plasma. Morinda officinalis capsules also enhanced the bone strength and prevented the deterioration of trabecular microarchitecture. CONCLUSION Morinda officinalis capsules possess potent anti-osteoporotic activity in OVX rats which could be an effective treatment for postmenopausal osteoporosis.
Acid Phosphatase ; blood ; Alkaline Phosphatase ; blood ; Animals ; Bone Density ; drug effects ; Bone Density Conservation Agents ; pharmacology ; therapeutic use ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Humans ; Isoenzymes ; blood ; Morinda ; Osteocalcin ; blood ; Osteoporosis, Postmenopausal ; blood ; metabolism ; prevention & control ; Ovariectomy ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase

OBJECTIVETo compare inhibitory effects of recombinant receptor activator of nuclear factor kappaB protein with bisphosphonate treatment (ALN) on osteoclasts activity and bone loss in ovariectomized mice.

METHODSTwenty-four female KM mice were ovariectomized bilaterally and treated with recombinant receptor activator of nuclear factor kappaB protein, alendronate, or PBS. Twelve weeks later, body weight, biochemical markers of bone metabolism, Micro CT scan and bone morphology were examined.

RESULTSAfter 12 weeks administration, the Micro CT scan and bone morphology values of each group were as follow. The control group: BMD (92.600 +/- 14.319) mg/cc, Tb.Th (0.094 +/- 0.011) mm, Tb.Sp (0.455 +/- 0.124) mm, BVF 0.192 +/- 0.023, SMI 1.388 +/- 0.328; the recombinant receptor activator of nuclear factor kappaB protein group: BMD (133.050 +/- 13.022) mg/cc, Tb.Th (0.098 +/- 0.009) mm, Tb.Sp (0.365 +/- 0.105) mm,BVF (0.291 +/- 0.025)%, SMI 0.661 +/- 0.384; the ALN group: BMD(128.013 +/- 16.040) mg/cc, Tb.Th (0.097 +/- 0.011) mm, Tb.Sp (0.376 +/- 0.104) mm, BVF 0.281 +/- 0.024, SMI 0.753 +/- 0.307. In the ovariectomized mice experiments, both recombinant receptor activator of nuclear factor kappaB protein and ALN significantly inhibited ovariectomy-induced bone loss. Compared to the control group (PBS), the recombinant receptor activator of nuclear factor kappaB protein group showed increased distal femur BMD and decreased trabecular spacing (Tb.Sp), whereas the control group had significantly decreased distal femur BMD, significantly decreased Tb.Th, and increased Tb.Sp. There was a significant difference in bone volume fraction among the groups. The TRAP-positive osteoclasts in distal femur bone slices were nearly complete inhibited for Recombinant receptor activator of nuclear factor kappaB protein group and alendronate group.

CONCLUSIONIn vivo, recombinant receptor activator of nuclear factor kappaB protein effectively inhibits the activity of osteoclasts and the resulting bone loss, which has a similar effect as alendronate.

Animals ; Bone Density ; drug effects ; Diphosphonates ; therapeutic use ; Female ; Mice ; Osteoporosis ; diagnostic imaging ; drug therapy ; Ovariectomy ; Radiography ; Receptor Activator of Nuclear Factor-kappa B ; therapeutic use

Diagnosis, Differential ; Female ; Humans ; Middle Aged ; Pancreatic Neoplasms ; diagnosis ; Solitary Fibrous Tumors ; diagnosis

Objectives To investigate gestational multiple metabolic abnormalities aggregation and diagnostic criteria for gestational metabolic syndrome(GMS),and to analyze the risk factors of GMS.Methods A cohort study recruiting 309 pregnant women with preeclampsia,627 pregnant women with gestational diabetes mellitus(GDM)and 1245 normal pregnant women was performed from January 2008 to December 2011 in Guangdong Women and Children's Hospital.Information regarding age,gestational weeks,basic blood pressure,admission blood pressure,height and body mass index(BMI)before pregnancy was recorded.Biochemical indicators including fasting plasma glucose(FPG),fasting insulin (FINS),total cholesterol(TC),triglyceride(TG),high density lipoprotein(HDL-C),low density lipoprotein(LDL-C),free fatty acids(FFA)were tested.GMS was diagnosed with three or all of the following conditions:(1)overweight and/or obesity before pregnancy(BMI ≥ 25 kg/m2);(2)hypertension with blood pressure ≥ 140/90 mm Hg(1 mm Hg =0.133 kPa);(3)hyperglycemia:diagnosed as GDM;(4)dyslipidemia with TG≥3.23 mmol/L The incidence of GMS of the three groups were calculated and the risk factors were analyzed.Results(1)The age,gestational weeks,basic blood pressure,admission blood pressure,BMI before pregnancy of women with preeclampsia and women with GDM were significantly different compared to normal women,respectively(P ＜ 0.01).(2)Biochemical indicators of women with preeclampsia were as following:FPG(4.6 ± 1.0)mmol/L,FINS(10.1 ± 5.6)mU/L,TC(6.3 ±1.6)mmol/L,TG(3.9 ± 1.8)mmol/L,HDL-C(1.4 ±0.4)mmol/L,LDL-C(3.0 ± 1.0)mmol/L,FFA (0.8 ±0.4)mmol/L.And those in women with GDM were:FPG(4.7 ± 0.9)mmoL/L,FINS(10.2 ± 5.8)mU/L,TC(5.7 ± 1.3)mmol/L,TG(3.2 ± 1.1)mmol/L,HDL-C(1.4 ± 0.4)mmol/L,LDL-C (2.7 ± 0.9)mmol/L,FFA(0.6 ± 0.3)mmol/L In normal pregnant women they were:FPG(4.3 ±0.5)mmol/L,FINS(9.0±4.4)mU/L,TC(5.7 ±1.1)mmol/L,TG(2.8 ±1.1)mmol/L,HDL-C (1.5 ± 0.4)mmol/L,LDL-C(2.9 ± 0.8)mmol/L,FFA(0.6 ± 0.2)mmol/L Statistic differences were found in preeclampsia and GDM women compared to normal women respectively(P ＜ 0.01).(3)The prevalence of GMS in preeclampsia group and in GDM group was 26.2％(81/309)and 13.6％(85/627),statistically different from that of the control group(0)(P ＜0.01).(4)Compared to normal women,women with preeclampsia had higher risk of developing GMS(OR =1.62,95 ％ CI 1.31-2.00,P ＜ 0.01).The risk factors were BMI(OR =1.29,95％ CI 1.13-1.47)and TG(OR =2.49,95％ CI 1.87-3.31).Also,women with GDM had higher risk of developing GMS than normal women(OR =1.27,95％ CI 1.09-1.49,P ＜ 0.01),and the risk factors were BMI(OR =1.13,95 ％ CI 1.04-1.23)and TG(OR =1.16,95 ％ CI 1.02-1.33).TG was the independent risk factor in both preeclampsia women and GDM women(P ＜ 0.01,P ＜ 0.05).HDL-C seemed to have less importance in identifying GMS(P ＞ 0.05).Conclusions According to the GMS diagnostic criteria used in this study,some preeclampsia patients and some GDM women had aggregation of multiple metabolic abnormalities including pre-pregnancy overweight/obesity,hyperglycemia,high blood pressure and dyslipidemia.TG was the independent risk factor for GMS.HDL-C seemed to have less importance in identifying GMS.

Objective To evaluate the utility of MMP9,MPO and sCD40L in detection of the character of coronary artery plaque.Methods From April 2008 to January 2010,118 patients from outpatient of Fu Wai Hospital with chest pain were enrolled.All of them underwent 64 Multiple-detector row spiral computer tomography (64-MDCT),the CT value ＜ 130 Hu patients were enrolled in non-calcified plaque group (71 cases),CT value ≥ 130 Hu patients were enrolled in the calcified plaque group (47 cases).Ninty healthy volunteers were selected as the control group.Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum markers,including MMP9,MPO and sCD40L.Levels of MMP9,MPO and sCD40L of each group were compared.ROC curve was used to evaluate the sensitivity and specificity of the markers in diagnosis of non-calcified plaque.Results MMP9,MPO and sCD40L levels of non-calcified were ( 762.25 ± 368.71 ),[ 844.10 (582.00 - 1220.70) ],(9.37 ± 3.15) μg/L,higher than the healthy control group (342.70 ± 178.53),[426.35 ( 283.20 - 592.00) ],(6.55 ± 2.96) μg/L and calcified plaque group ( 483.12 ± 219.09 ),[ 469.00 ( 302.45 - 723.55) ],( 7.24 ± 2.86) μg/L The difference was statistically significant ( F =42.47,H =50.28,F =17.94,all P ＜ 0.01 ). Areas of MMP9,MPO and sCD40L under the ROC curve to predict non-calcified plaque were 0.854,0.792,0.751 respectively,when the identification threshold for non-calcified plaque were 510.13,537.82,7.05 μg/L respectively,the diagnostic sensitivity was 80％,80％,80％ respectively,and specificity was 80％,67％ and 55％ respectively.Conclusion The serum MMP9,MPO and sCD40L levels can help to determine the　character of coronary plaque.

Objective To detect the value of serum galactomannan (GM) for diagnosis of invasive aspergillosis (IA) in hematological patients. Methods We prospectively evaluated the diagnostic value of twice-weekly screening for circulating Aspergillus fumigatus with sanditch enzyme linked immunosorbent assay. Results On the basis of the analysis of 472 serum samples from 113 episodes of 92 patients, the sensitivity, specificity, positive and negative predictive values of the sandwich ELISA for proven and probable IA cases were 83.3 %, 91.1 %, 78.9 % and 93.1 %, respectively, when samples with 2 consecutive positive results≥0.7 were used. Furthermore, GM antigenemia was detected before the onset of radiologic signs with a median of 7 days (range, 1-14 days), before isolation of Aspergillus with a median of 4 days (range, 1-7 days),and before anti-fungal therapy with a median of 6 days (range, 1-15 days). Conclusion The sandwich ELISA for GM detection is a reliable method for early diagnosis of IA in patients with haematological diseases.

Objective To evaluate the value of MSCT, MRI and SPECT in detecting acute myocardial ischemia in Chinese mini-swine model. Methods A total of six male mini-pigs were recruited with a mean body weight of (21.6 ± 1.2) kg. All pigs were scanned on MSCT before the ligation of distal segment of Left anterior descending artery. Then, MSCT was rescanned every 2 h from ligation till 8 h latter.MRI, SPECT and the last MSCT scan were performed within 24 h one by one. Finally pathological examination was carried out right after the pig killed. Results One pig died during operation, the other 5 finished all the examinations. The pathological staining showed the same areas of myocardial infarction in the left ventricular anterior wall with the all the imaging findings, including low perfusion region of MSCT arterial phase at 2-24 h, low perfusion region of SPECT at 24 h and low perfusion region of MRI first pass phase at 24 h. Three of 5 pigs showed enhanced edge of low perfusion region on MSCT delayed scan at 4-8 h. The mean CT values in the region with reduced first-pass perfusion were 75.9,36.4, 35. 2,37. 8,37.4,33.3 HU on MDCT image at baseline, 2-8 h after operation and within 24 h after operation, respectively,and there were statistically significant difference of CT values ( F = 12. 341, P ＜0. 01 ) between preoperative and all postoperative MSCT scan. There were no statistically significant difference (F = 2. 278, P = 0. 792)among all postoperative MSCT scan. At baseline, 2-8 h after operation and within 24 h, the average volumes of stroke volume(SV)were 21.7,11.9,10.3,11.4,12. 3,12.6 ml, respectively, while the average volume of end-systolic volume( ESV)were 15.2,23.4,25.0,24. 4,25.3,22. 8 ml,respectively. The average volume of end-diastolic volume ( EDV ) at these time point were 37. 0,35.4,35.0,35.7,37. 6,37.5 ml,respectively and the average percentage of ejection fraction (EF) were 58.9% ,33.8% ,29. 0%, 31.9%,32.6% ,33.5% ,respectively. SV(F =22. 349, P＜0.01) ,ESV (F=8. 810, P＜0.01) ,EF(F =27. 240,P ＜ 0. 01 ) were all significantly different among all postoperative MSCT scan except EDV ( F = 2. 339, P =0. 079). Infarct size, which was defined as the proportion of the area of infarction to that of the entire heart,were (39.4 ±12.6)% for MSCT,(37.2 ± 10.0)% for MRI, (35.9 ±9.6)% for TTC, respectively.There were no significant differences of infarct size between TTC and MSCT (t =0. 612, P =0. 574), TTC and MRI (t=0.820, P=0.458), MSCTand MR (t=0. 425 ,P =0. 692 ). Conclusions MSCT,MRI and SPECT were all able to be used to detect the myocardial infarction in acute myocardial ischemia model The infarct size defined on MSCT, MRI and pathology were consistent. The density of ischemic myocardium and cardiac function did not change over the time within 24 h right after infarction.

Objective To study the correlation between morphological characteristics of coronary artery calcified plaques and luminal stenosis of local coronary artery segments with 64-slice computed tomography.Methods One hundred and eleven patients who had undergone 64-slice computed tomographic coronary angiography(CTCA)and conventional coronary angiography(CAG)were retrospectively analyzed.The calcified plaques were classified as punctate,nodular,strip-like and nubbly in long-axis view of coronary artery lumen,and were classified as crescent,semilunar,round moon and circinate in short-axis view.The morphologic characteristics of these calcified plaques on CTCA were retrospectively analyzed and compared with luminal stenosis of CAG results.Results Among 528 calcified coronary segments which were analyzed in 111 patients,there were 383(72.5%)punctate calcified plaque segments and 145(27.5%)of non-punctate plaques.There were 34(23.4%,34/145)non-punctate calcified plaques which caused severe stenosis(≥75%),including 4(11.8%)nodular,8(23.5%)stripe-like and 22(64.7%)nubbly calcified plaques on the long-axis view,and 0(0.0%)cresent,8(23.5%)semilunar,18(52.9%)round moon and 8(23.5%)circinate calcified plaques on the short-axis view.The ratios of different morphological coronary artery calcifications which caused severe stenoses were significantly different with each other(all P＜0.01).Conclusion Different figures of coronary artery calcified plaques demonstrate different degrees of stenoses of local coronary artery lumen.Severe stenoses were mostly caused by nubbly calcified plaque on long-axis view,round moon and circinate calcified plaque on short-axis view.