ADP-ribosylhydrolases: from DNA damage repair to COVID-19 / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Adenosine diphosphate (ADP)-ribosylation is a unique post-translational modification that regulates many biological processes, such as DNA damage repair. During DNA repair, ADP-ribosylation needs to be reversed by ADP-ribosylhydrolases. A group of ADP-ribosylhydrolases have a catalytic domain, namely the macrodomain, which is conserved in evolution from prokaryotes to humans. Not all macrodomains remove ADP-ribosylation. One set of macrodomains loses enzymatic activity and only binds to ADP-ribose (ADPR). Here, we summarize the biological functions of these macrodomains in DNA damage repair and compare the structure of enzymatically active and inactive macrodomains. Moreover, small molecular inhibitors have been developed that target macrodomains to suppress DNA damage repair and tumor growth. Macrodomain proteins are also expressed in pathogens, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, these domains may not be directly involved in DNA damage repair in the hosts or pathogens. Instead, they play key roles in pathogen replication. Thus, by targeting macrodomains it may be possible to treat pathogen-induced diseases, such as coronavirus disease 2019 (COVID-19).

A case of infantile occult primary intestinal lymphangiectasia / 中国综合临床

Objective:In order to improve the understanding of the clinical, endoscopic and pathological characteristics of occult primary intestinal lymphangiectasia in infants, and to reduce misdiagnosis and missed diagnosis, we analyzed the clinical manifestations and diagnosis and treatment related data of a case of primary intestinal lymphangitis in Xiamen Children′s Hospital.The patient was admitted to the hospital with bronchopneumonia, mild diarrhea and edema, without decreasing in the number of peripheral blood lymphocytes, showing hypoproteinemia.Albumin infusion failed to alleviate hypoalbuminemia, and then gastroscopy showed that the duodenal segment had white millet like granular protrusion with different sizes and dense density, and the mucosa had extensive leukoplakia like lesions.Pathological examination showed that the small intestinal lymphadenectasia was obvious, and then the small intestinal lymphadenectasia was confirmed.After diagnosis, albumin was infused every two days for two consecutive courses and portagen medium chain fatty acid milk powder was fed.After treatment, the total protein and albumin in the plasma basically rose to the normal level, and the color Doppler ultrasonography of abdomen and chest showed that both pleural effusion and bilateral pulmonary effusion had been absorbed.The retrospective analysis of this case shows that for infants with dropsy caused by the decrease of plasma albumin and globulin, the possibility of excessive protein consumption caused by liver and kidney disease, tumor, tuberculosis and other chronic diseases should be excluded, and small intestinal lymphadenopathy should be considered, gastroscopy should be performed as soon as possible, pathological biopsy should be further improved, early diagnosis and treatment should be carried out as soon as possible And diet intervention treatment to avoid serious consequences caused by not timely handling.

Signaling pathway of cardioprotection induced by monophosphoryl lipid A in rabbit myocardium.

OBJECTIVE: To investigate the effect of monophosphoryl lipid A (MLA) on ischemia/reperfusion (I/R) injury in rabbit heart and its signal transduction pathway. METHODS: Eighteen rabbits were randomly assigned to three groups: (1) MLA-group: Six rabbits received injection of MLA (35 &mgr;g/kg, iv) 24 h prior to 45 min of ischemia followed by 60 min of reperfusion. (2) Control group: Six rabbits received an intravenous bolus injection of the same volume of vehicle. (3) SB group: Six rabbits intravenously received 2 &mgr;mol/l SB 203580 30-min prior to MLA administration. At the end of reperfusion, myocardial infarct size and serum nitrite/nitrate were detected. Myocardium p38 mitogen-activated protein kinase (p38 MAPK) was detected using a Western blotting method. RESULTS: MLA pretreatment caused a significant reduction of infarct size as percent of area at risk as compared to vehicle-pretreated controls (36+/-3 vs. 65+/-4%, P<0.001). Total nitrite in serum increased in MLA pretreated animals (P<0.05). The protective effects of MLA were completely abolished by selective p38 MAPK inhibitor SB 203580. Western blot analysis showed significant accumulations of p38 MAPK proteins in the myocardium of MLA pretreated animals. CONCLUSIONS: MLA can protect the heart by reducing myocardial infarct size in rabbits. This cardioprotective effect might be attributed to upregulation of nitric oxide (NO) through the activation of p38 MAPK.

Study on HPLC Fingerprint of Qingbi Granules / 中国药房

OBJECTIVE:To establish HPLC fingerprint of Qingbi granules.METHODS:HPLC method was adopted.The determination was performed on Diamonsil C18 column with mobile phase consisted of methanol-0.3％ phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 260 nm,and column temperature was 25 ℃.The sample size was 10 μL.Using baicalin as reference,HPLC chromatograms of 10 batches of samples were determined.Common peak identification and similarity evaluation were performed by using TCM Chromatographic Fingerprint Similarity Evaluation Software (2.0 edition).RESULTS:There were 29 common peaks in HPLC chromatograms of 10 batches of samples.The similarity among the 10 batches was more than 0.90.After validation,HPLC chromatograms of 10 batches of samples were in line with control fingerprints.CONCLUSIONS:Established fingerprints can provide reference for identification and quality evaluation of Qingbi granules.

Effect of EGCG-Zn on the Learning,Memory and Antioxidant Abilities in Vascular Dementia Rats / 中国药房

OBJECTIVE:To study the effect of EGCG-Zn chelated by epigallocatechin gallate (EGCG) and Zn ion on learn-ing,memory and antioxidant abilities in vascular dementia(VD)rats. METHODS:Rats were randomly divided into sham group, model group,zinc gluconate group (positive control,70 mg/kg),EGCG group (5 mg/kg),and EGCG-Zn low-dose,mid-dle-dose,high-dose groups (2,5,10 mg/kg),10 in each group. The VD models were induced by middle cerebral artery occlu-sion. The rats were intragastrically given relevant medicines after 7 d of modeling,once a day,for 28 d. Morris water maze were used to evaluate learning and memory abilities of rats. Spectrophotometric method was adopted to detect the superoxide dismutase (SOD),catalase(CAT),glutathione peroxidase(GSH-Px)activities and malondialdehyde(MDA)content in brain tissue of rats. RESULTS:Compared with sham group,escape latency was extended in model group,percentage of platform quadrant swimming distance(dP/dT)was reduced(P<0.01);SOD,CAT,GSH-Px activities in brain tissue were reduced,and MDA content was in-creased (P<0.01). Compared with model group,escape latency on 4th d of recording was obviously shortened in EGCG group and EGCG-Zn dose groups,dP/dT was obviously increased and dose-depended(P<0.05 or P<0.01);SOD,CAT,GSH-Px activi-ties in brain tissue were increased,MDA content was decreased(P<0.05 or P<0.01)and dose-depended. Compared with EGCG group and zinc gluconate group,escape latency was obviously shortened in EGCG-Zn dose groups,dP/dT was increased;SOD, CAT,GSH-Px activities in brain tissue were increased,MDA content was decreased;and the effects in high-dose,medium-dose groups were more obvious(P<0.05 or P<0.01). CONCLUSIONS:EGCG-Zn can improve the learning,memory and antioxidant abilities in VD rats,and the effect is superior to EGCG.

Effects of Luteolinl on the Proliferation and Differentiation in Human Periodontal Ligament Cells / 沈阳医学院学报

Objective:To evaluate the biological effects of Luteolinl on the proliferation and differentiation in human periodontal ligament cells (PDLC) . Methods:MTT, ALP kit and Q-PCR was used to detect the expression of osteogenesis related gene. Results:Luteolinl (100, 10, 1, 0.1, 0.01μmol/L) could increase the proliferation of PDLC, and there were significant differences compared with control group (P<0.05) . The ALP Kit results showed that Luteolinl could increase the ALP actiuty of PDLC (P<0.05) . The Q-PCR results showed that Luteolin could increase the expression of ALP and RUNX2 (P<0.05) . Conclution:At proper concentration,Luteolinl can increase the proliferation and differentiation of PDLC.

Research progress on curcuma zedoary extracts inhibiting vascular smooth muscle cell proliferation in pathological process of in-stent restenosis / 中国药理学通报

Vascular smooth muscle cells (VSMCs) proliferation is the critical pathological process of in-stent restenosis (ISR). Recent researches indicate that the traditional Chinese medicine curcuma zedoary (E’zhu) can inhibit VSMCs proliferation, with a potential value in preventing and treating ISR. The major ac-tive components of curcuma zedoary are curcuma and β-elemene. The underlying mechanisms involve the inhibition of hemeoxygen-ase-1 expression, blockade of extracellular signal-regulated ki-nase – MAPK and Akt pathways, and subsequent cell cycle ar-rest. This article reviews the recent progress on curcuma zedoary extracts regulating VSMCs proliferation in ISR.

Study on Quality Standards for Compound Shatagan Oral Liquid / 中国中医药信息杂志

Objective To establish the quality standards for compound Shatagan Oral Liquid. Methods Chuanxiong Rhizoma was identified by TLC. The content of ferulic acid was determined by HPLC. Separation was performed on a Diamonsil C18 column (4.6 mm × 250 mm, 5μm) with a mobile phase consisting of methanol-1% acetic acid solution in gradient elution (0-5 min, 35% methanol;5-8 min, 35%→23% methanol;8-22 min, 23% methanol) at 30℃;The flow rate was 1.0 mL/min;The injection volume was 5μL;The detection wavelength was 322 nm.Results Ferulic acid showed a good linear relationship in the range of 0.039 4-0.630 0μg (r=0.999 7,n=7). The average recovery was 98.22% and RSD was 2.62% (n=6).Conclusion The method is reliable, sensitive and with repeatability, which can be used as the quality control method for compound Shatagan Oral Liquid.

Effects of resveratrol on blood pressure and cardiac function in the rats with vascular calcification / 中国比较医学杂志

Objective To observe the effects of resveratrol on blood pressure and cardiac function in the rats with vascular calcification induced by vitamin D3 plus nicotine.Methods 32 male SD rats were randomly divided into the following groups for treatment:Control (Con), calcified (Cal), Cal+Res low dose [L] and Cal+Res high dose [H] groups.Blood pressure, cardiac function, alkaline phosphatase ( ALP ) activity in serum or aorta were detected and HE staining was used for pathological examination at 6 weeks after treatment.Results Compared with the Con group, LVW/BW, heart rate, systolic aortic pressure, pulse pressure, mean blood pressure, left ventricular systolic pressure and ALP activity in serum or aorta of rats in the Cal group were increased by 27.3%, 8.8%, 22.8%, 47.5%, 13.6%, 19.0%, 280%and 265%( P<0.05 or P<0.01) , respectively.Compared with the Cal group, pulse pressure and ALP activity in serum or aorta of rats in the Cal+Res [L] group were decreased by 8.5%, 34.5%and 29.5%(P<0.05 or P<0.01), respectively, and LVW/BW, systolic aortic pressure, pulse pressure, mean blood pressure, left ventricular systolic pressure and ALP activity in serum or aorta of rats in the Cal +Res [ H] group were decreased by 14.2%, 13.6%, 23.7%,10.0%, 9.0%, 53.1%and 45.9%(P<0.05 or P<0.01), respectively.Compared with the Cal+Res [L] group, systolic aortic pressure, pulse pressure, left ventricular systolic pressure and ALP activity in serum or aorta of rats in the Cal+Res [H] group were decreased by 8.3%, 16.7%, 5.8%, 28.4% and 23.2% (P<0.05 or P<0.01), respectively.Compared with the aorta in the Con group, pathological examination revealed thickened vessel walls and disordered elastic fibers in the calcified aortas.However, the thickness of aortic wall in the Cal+Res [ L] and Cal+Res [ H] groups was reduced and elastic fibers were regularly arranged.Conclusion Resveratrol can effectively reduce the blood pressure and improve the cardiac function in rats with vascular calcification.

Experimental study of iontophoresis-mediated corneal collagen crosslinking for rabbit Staphylococcus aureus keratitis / 中华实验眼科杂志

Background It is determined that riboflavin/ultraviolet A (UVA)-induced corneal collagen crosslinking is able to increase resistance of cornea against enzymatic digestion and has antimicrobial efficacy for various kinds of bacteria in vitro.However,its in vivo study is less now.Objective This study aimed to evaluate the efficacy of iontophoresis-mediated corneal collagen crosslinking combined with or without drugs for Staphylococcus aureus keratitis.Methods Bacterial keratitis models were induced by the interstromaly injection of Staphylococcus aureus suspension with concentration 2× 109/ml in the right eyes of 40 rabbits,and then the rabbits were randomly classified into the model group,gatifloxacin eye drops group,riboflavin/UVA corneal crosslinking group and drugs+ crosslinking group.The smearing of corneal surface was performed for the identification of bacteria 24 hours after injection.Iontophoresis-mediated riboflavin/UVA crosslinking was applied on the eyes of the riboflavin/UVA corneal crosslinking group and drugs+crosslinking group,and gatifloxacin eye drops was topically used 7 times per day on the eyes of the gatifloxacin eye drops group and drugs+crosslinking group.The corneal inflammation was examined and graded under the slit lamp biomicroscope before and after treatment.Ocular anterior segment optical coherence tomography(AS-OCT),corneal histopathology and ultrastructure were examined 14 days after treatment.The living environment of the experimental animals was maintained at 21 ℃ with a 12-hour light and dark cycle.Animals used in this study were treated in accordance with the Weifang Medical College Animal Experimentation Ethic Committee (AEEC) guidelines.The study protocol was approved by the AEEC.Results Corneal inflammation and ulcer were observed,but no significant difference was found in the inflammatory grade among the 4 groups 24 hours after injection (x2=0.293,P＞0.05).In the 14th day after injection,the corneal ulcer area was smaller and corneal edema was milder in the drugs+crosslinking group compared with the model group,gatifloxacin eye drops group and riboflavin/ UVA corneal crosslinking group,showing a significant difference in the inflammatory grade among them (x2 =38.710,P＜0.001).The cornea thickness values of ulcer zone were (428.1 ± 146.2) μm on the 14th postinjected day in the drugs+crosslinking group,which was evidently higher than those in the model group,gatifloxacin eye drops group and riboflavin/UVA corneal crosslinking group,with a significant difference among the 4 groups (F =8.310,P＜0.001).A lower degree of destruction of cornea collagen and less inflammatory cells were seen in the cornea tissue of the drugs+ crosslinking group by haematoxylin and eosin staining in comparison with other 3 groups,and normal keratocytes were much more in the drugs + crosslinking group than those in other treated groups.Conclusions Iontophoresismediated corneal collagen crosslinking can alleviate Staphylococcus aureus keratitis.The combination of crosslinking with drugs has a better effectiveness than the administration of gatifloxacin eye drops only or riboflavin/UVA corneal crosslinking only.

Effects of Panax quinquefolium saponin on mitochondrial membrane po-tential and cardiomyocyte apoptosis in ischemia/reperfusion rats / 中国病理生理杂志

AIM:To investigate whether mitochondrial membrane potential (ΔΨm ) and the mitochondrial ap-optotic pathway are involved in the protective mechanism of Panax quinquefolium saponin ( PQS) against cardiomyocyte ap-optosis after ischemia/reperfusion ( I/R) injury in rat myocardium .METHODS: Ninety healthy male SD rats were ran-domly divided into sham group , I/R group, PQS (200 mg· kg -1 · d-1 ) +I/R group, cyclosporine A ( CsA) group, CsA (10 mg· kg-1 ) +I/R group and PQS +CsA +I/R group.The model of myocardial I/R injury in vivo was established by ligating the left anterior descending artery ( LAD) for 30 min followed by 120 min of reperfusion in the rats .The serum ac-tivity of lactate dehydrogenase ( LDH ) was measured by automatic chemistry analyzer .The myocardial infarct size was measured by Evans blue and 2,3,5-triphenyltetrazolium chloride (TTC) staining.Cardiomyocyte apoptosis was detected by in situ TDT-mediated dUTP nick end labeling (TUNEL).The protein levels of Bcl-2, Bax, cleaved caspase-3 and cytosol-ic cytochrome C were determined by Western blotting .ΔΨm was measured by laser scanning confocal microscopy and fluo-rescence microplate reader .RESULTS:Compared with I/R group, the serum content of LDH ,the infarction size in PQS+I/R group, CsA+I/R group and PQS +CsA+I/R group and the myocardial apoptotic index were decreased .Compared with I/R group, the fluorescence intensity of mitochondria after JC-1 staining was enhanced in PQS +I/R group, CsA+I/R group and PQS+CsA+I/R group, and the relative fluorescence units (RFU) ofΔΨm were improved in those 3 groups. In PQS+I/R group, CsA+I/R group and PQS+CsA+I/R group, the protein expression of Bcl-2 was increased, and that of Bax was decreased compared with I/R group.Moreover, in those 3 groups, the protein levels of cleaved-caspase-3 and cytosolic cytochrome C were decreased compared to I /R group, respectively.CONCLUSION:PQS attenuates myocardial injury and cardiomyocyte apoptosis during I /R, and the protective mechanisms of PQS were associated with the modulation ofΔΨm and the inhibition of mitochondrial apoptosis pathway .

PQS attenuates cardiomyocyte apoptosis induced by thapsigargin through inhibiting endoplasmic reticulum stress / 中国病理生理杂志

AIM:To study the effect of Panax quinquefolium saponin (PQS) on cardiomyocyte apoptosis in-duced by thapsigargin ( TG) .METHODS:Primary cultured cardiomyocytes from neonatal SD rats were divided into con-trol group, TG group, PQS (40 mg/L, 80 mg/L and 160 mg/L) +TG group, si-PERK+TG group, and mock+TG group.The cells were treated with 1 μmol/L TG for 24 h to induce apoptosis.The PERK gene in the cardiomyocytes was knocked down by RNAi.The cell viability was detected by CCK-8 assay.Apoptosis was analyzed by flow cytometry.Wes-tern blotting was used to determine the expression of ERS molecules GRP78, CRT, ATF4 and CHOP, anti-apoptosis pro-tein Bcl-2 and pro-apoptosis protein Bax.RESULTS: Compared with control group, TG significantly and the apoptosis, reduced the cell viability (P<0.05), increased the phosphorylation of PERK and eIF2α, increased the expression of GRP78, CRT, ATF4, CHOP and pro-apoptosis protein Bax, and decreased the expression of anti-apoptosis protein Bcl-2 (P<0.05).Compared with TG group, PQS treatment (160 mg/L) significantly reduced the apoptosis and increased the cell viability (P<0.05).All the 3 different concentrations of PQS significantly increased the expression of anti-apoptosis protein Bcl-2 and reduced the expression of pro-apoptosis protein Bax (P<0.05) in a dose-dependent manner.PQS pre-treatment and knockdown of PERK both reduced the protein levels of GRP78, CRT, PERK, p-PERK, eIF2α, p-eIF2α, ATF4, CHOP and pro-apoptosis protein Bax, and increased the expression of anti-apoptosis protein Bcl-2 (P<0.05). CONCLUSION:PQS at concentration of 160 mg/L attenuated cardiomyocyte apoptosis induced by TG.PQS had the simi-lar effect as PERK knockdown on cardiomyocyte apoptosis.The mechanism may be associated with inhibiting PERK-eIF2α-ATF4-CHOP pathway of ERS-related apoptosis.

The diagnostic significance of nailfold videocapillaroscopy in systemic sclerosis / 中华内科杂志

ObjectiveTo observe nailfold capillary changes in a cohort of conncctive tissue disease ( CTD ) with Raynaud's phenomenon (RP) and to explore the diagnostic value of nailfold videocapillaroscopy (NVC) in systemic sclerosis(SSc).MethodsSixty CTD patients with RP divided into SSc group (n =36) and non-SSc group ( n =24) were referred to an experienced operator for NVC.ResultsThe patients had decreased capillary loops in SSc group with the capillary diameter more enlarged in SSc group than nonSSc group.The number of patients in SSc group with giant capillaries was 14,while 3 in non-SSc group.There were 23 patients with haemorrhagcs in SSc group and 9 in non-SSc group.The number of patients with severe effusion was 15 in SSc group,while 2 in non-SSc group.By using the ROC curves,indexes with AUC at least 0.7 of the input capillary diameter,the output capillary diameter,the middle capillary diameter,blood color and effusion for the diagnostic cutoff points were 18.5 μm,24.5 μm,19.5μm,deep red and severe effusion.With at least 2 out of the top 3 indexes,the diagnostic sensitivity and specificity of SSc were higher.ConclusionsCTD Patients with RP of SSc have less capillary loops,more enlarged capillaries,more giant capillaries, moresevereeffusionandmorehaemorrhagesthannon-SScpatients. The characteristics of nailfold capillary changes in SSc patients with RP can be helpful tor the diagnosis and the differential diagnosis of SSc.

Study on chemical constituents of Artemisia sphaerocephala / 中国中药杂志

Further study on the chemical constituents of Artemisia sphaerocephala led to the isolation of fifteen compounds. These compounds were isolated and purified by repeated column chromatography on silica gel and Sephadex LH-20. Their structures were determined by physicochemical properties and spectral data analysis. These compounds were identified as 5-hydroxy-7,4'-dimethoxyflavanone (1), 5-hydroxy-7, 4'-dimethoxyflavone (2), 5-hydroxy-7, 4'-dimethoxyflavanonol (3), 5,3'-dihydroxy-7, 4'-dimethoxyflavanone (4), 5,7-dihydroxy-6,4'-dimethoxyflavone (5), isosakuranetin (6), hesperetin (7), navingenin (8), acacetin (9), chrysoeroil (10), 5,7-dihydroxy-4'-methoxyflavone-6,8-di-C- -glucopyranoside (11), didymin (12), acacetin-7-O-rutinoside (13), piceine (14), and capillarin (15). Compounds 1-3, 5, 7, 8, 10-15 were isolated from this plant for the first time.

MicroRNA-15a/b are up-regulated in response to myocardial ischemia/reperfusion injury / 老年心脏病学杂志（英文版）

Objective Several studies have indicated that miR-15a,miR-15b and miR-16 may be the important regulators of apoptosis.Since attenuate apoptosis could protect myocardium and reduce infarction size,the present study was aimed to find out whether these miRNAs participate in regulating myocardial ischemia reperfusion (I/R) injury.Methods Apoptosis in mice hearts subjected to I/R was detected by TUNEL assay in vivo,while flow cytometry analysis followed by Annexin V/PI double stain in vitro was used to detect apoptosis in cultured cardiomyocytes which were subjected to hypoxia/reoxygenation (H/R).Taqman real-time quantitative PCR was used to confirm whether miR-15a/15b/16 were involved in the regulation of cardiac I/R and H/R.Results Compared to those of the controls,I/R or H/R induced apoptosis of cardiomyocytes was significantly iucreased both in vivo (24.4％ ± 9.4％ vs.2.2％ ± 1.9％,P ＜ 0.01,n =5) and in vitro (14.12％ ±0.92％ vs.2.22％ ± 0.08％).The expression of miR-15a and miR-15b,but not miR-16,was increased in the mice I/R model,and the results were consistent in the H/R model.Conclusions Our data indicate miR-15 and miR-15b are up-regulated in response to cardiac I/R injury,therefore,down-regulation of miR- 15a/b may be a promising strategy to reduce myocardial apoptosis induced by cardiac I/R injury.

Biocompatibility of silk scaffold/3T3-L1 preadipocytes compounds:An in vitro experiment / 中国组织工程研究

BACKGROUND: Silk, a natural product, has been paid wide attention in medical field owing to a fact that silk's mechanical property and biocompatibility are superior to traditional artificially synthesized degradable polymers. OBJECTIVE: To observe the effects of silk on the adsorption as well as cell morphology and function of 3T3-L1 preadipocytes. METHODS: Raw silk and trypsin-digested silk were ad Ubitum twisted into reticular fibrous cord with three-dimensional structure: mesh size 70-200 μm, thickness 200-300 μm, and porosity 20%. The three-dimensional scaffolds made of digested silk were placed in the 24-well plate. 3T3-L1 preadipocytes suspension (6×10~(10)/L) was added to the culture plate, 3 drops comprising 1 × 10~7 cells per well. Following 4-hour incubation in the air, when cells fully attached to the scaffold, they were thoroughly soaked with medium, which was renewed every other 2-3 days for a total of 1-4 weeks.RESULTS AND CONCLUSION: Inverted microscope results showed that silk scaffold/3T3-L1 preadipocytes compounds presented with slender prominences that stretched out and migrated ahead to gradually connect together through a head-to-end fusion fashion and enter into meshes. Scanning electron microscope results demonstrated that in the silk scaffold/3T3-L1 preadipocytes compounds, ceils tightly attached to silk scaffold, appropriately spread out, and secreted matrix. Silk shows better absorption for 3T3-L1 preadipocytes and maintains the normal morphology and functions of 3T3-L1 preadipocytes.

Accumulation and translocation of cadmium in soil and plant and its effects on growth of Artemisia annua and artemisinin content / 中国中药杂志

<p><b>OBJECTIVE</b>To study the accumulation and translocation of cadmium in the soil and Artemisia annua, and observe its effects on growth of A. annua and artemisinin content.</p><p><b>METHOD</b>A. annua were cultivated in pots with Cd concentration at 0.5, 1.5, 4.5 mg x kg(-1) level, respectively.</p><p><b>RESULT AND CONCLUSION</b>The growth of A. annua was inhibited at all the Cd levels characterized by the decreases of biomass and agronomic parameters; Most of Cd was accumulated in the roots of A. annua, and the ratios of Cd concentrations in roots and aerial part were 1.8:1 and 2.3:1 at 1.5, 4.5 mg x kg(-1) Cd level, respectively. Artemisinin content increased significant at 0.5 mg x kg(-1) Cd level, but there were no significant changes comparing with control group other Cd levels.</p>

Preparation and characterization of the antibodies against human myofibrillogenesis regulator 1 and application in neonatal rat cardiomyocytes / 中国病理生理杂志

AIM: To prepare and purify the polyclonal antibodies against human myofibrillogenesis regulator 1 (hMR-1), then to characterize the purity, titer, specificity and the availability.METHODS: Two polypeptides named peptide 1 and 2 were synthesized based on the bioinformatics analysis of the sequence of hMR-1 by using software TMHMM and DNAStar, then coupled with keyhole limpet hemocyanin (KLH) for immunization. These peptides for immunization were mixed and injected into New Zealand rabbits to prepare antibodies specifically against hMR-1. ELISA assay was used to detect the titers of the antibodies. After purification by immunoaffinity chromatography, antibodies were identified by Western blotting and immunocytofluorescent assays. Applications of the antibodies on neonatal rat cardiomyocytes were also employed.RESULTS: (1)The titers of antibodies were 1:10~5. In WB assay, a specific 17kD band was detected, corresponding to the predicted molecular weight of hMR-1; the positive fluorescent signals were distinct. (2)On the neonatal rat cardiomyocytes model, we observed a peri-nucleus location. The fluorescent signal of hMR-1 overexpression group was much stronger than that in vector control and normal control groups.CONCLUSION: All these results indicate that the antibodies obtained from poly peptides mixture immunization have either human original or rat original antigens. The antibody is available for using in Western blotting or immunofluorescent assays.

Resistant Mechanism and Strain Relationship of Multi-resistant Pseudomonas aeruginosa / 中华医院感染学杂志

OBJECTIVE To investigate the correlated resistant genes encoding ?-lactamases,aminoglycoside and genetic marker of integron and transposon in multi-resistant Pseudomonas aeruginosa(MRPA) isolated from clinical specimens and study their relationship by phylogenetic analysis.METHODS Twenty-one resistant genes,two integron-Ⅰ genes and one transposon genetic gene were analyzed by PCR and verified by DNA sequencing.Multi-resistant genes cluster analysis was performed by UPGMA.RESULTS The positive rates of CARB,oprD2,aac(3)-Ⅱ,aac(6′)-Ⅱ,ant(2″)-Ⅰ,intⅠ1,qacE△1-sul1 and merA in 20 strains of MRPA were 15%,100%,70%,15%,15%,85%,85% and 85%,respectively,and other genes were negative.It was classfied to three subgroup,by the multi-resistant genes cluster analysis.CONCLUSIONS MRPA isolated from clinical specimens has carried many resistant genes.The deficiency rate of oprD2 gene is very high.The positive rate of genetic mark genes about integron and transposon is very high.It may be the main multi-resistant mechanism of MRPA.Multi-resistant genes cluster analysis shows that there is clone transmission in MRPA and it can induce nosocomial infection prevalence.

Effect of insulin-like growth factor-1 on in vitro proliferation and differentiation of 3T3-L1 preadipocytes / 中国组织工程研究

BACKGROUND:Preadipocyte is a kind of specific precursor cells with the potential of proliferation and differentiation into adipocytes.If factor or drug with strong promotion of preadipocyte proliferation and differentiation is obtained and used in adipose tissue transplantation,which could elevate transplantation outcome of adipose tissues.OBJECTIVE:To find an optimal culture environment and to accumulate culture experience of in vitro culture 3T3-L1 preadipocytes,and to observe the effect of insulin-like growth factor(IGF)-1 on the proliferation and differentiation of 3T3-L1 preadipocytes,and to explore the optimal concentration of IGF-1.DESIGN,TIME AND SETTING:The cell control observation was performed at the Science Experiment Center,Liaoning University of Traditional Chinese Medicine from October 2007 to February 2008.MATERIALS:3T3-L1 preadipocytes were purchased from Shanghai Institutes For Biological Sciences.IGF-1 was bought from Biological,USA.METHODS:According to in vitro culture direction of 3T3-L1 preadipocytes,3T3-L1 preadipocytes were divided into 6 groups.3T3-L1 preadipocytes in the blank control group were inoculated in high-glucose DMEM supplemented with 0.1 volume fraction of fetal bovine serum.3T3-L1 preadipocytes in each experimental groups were respectively treated with high-glucose DMEM containing 5,10,20,50,100 ?g/L IGF-1 and 0.1 volume fraction of fetal bovine serum.MAIN OUTCOME MEASURES:Growth,proliferation and differentiation of 3T3-L1 preadipocytes in each experimental groups under the inverted microscope,and photographed.When cells were confluence,cell cycles were detected by flow cytometry.Growth rate of 3T3-L1 preadipocytes was measured by MTT essay.Fat growth rate in 3T3-L1 preadipocytes were examined by using oil red O stained-extracted assay.RESULTS:Inverted microscope showed that 3T3-L1 preadipocytes before differentiation were fusiform shape,with little fat in their endochylemas.3T3-L1 preadipocytes after differentiation were spherical shape,with much fat in their endochylemas.Lipid droplet gathered in the perinuclear space,and was stained orange by oil red O stained-extracted assay,forming ring-shape.Results from MTT essay and oil red O stained-extracted assay demonstrated that growth rate of 3T3-L1 preadipocytes and fat growth rate in 3T3-L1 preadipocytes were higher in each experimental groups treated with different doses of IGF-1 compared with the blank control group(P