Biopsy ; Cryopreservation ; Frozen Sections ; Humans ; Kidney ; pathology ; Kidney Diseases ; pathology ; Paraffin Embedding ; methods

BACKGROUND:After cerebral tissue ischemia and anoxia in young rats,the cerebral edema gets serious, and the levels of nitric oxide (NO) and malondialdehyde (MDA) decrease. Radix Astagali seu Hedysari has the pharmacological effects of enhancing immunity, anti-anoxia and improving myocardial ischemic reinfusion injury.OBJECTIVE: To investigate the influences of Radix Astagali seu Hedysari (huangqi) on contents of NO and MDA in brain tissue of young rats with cerebral injury after cerebral ischemia and anoxia.DESIGN:A randomized and controlled trial.SETTING: Department of Pediatrics, Second Hospital, Hebei Medical University; Department of Internal Medicine, Institute of Traditional Chinese Medicine, Hebei Medical University; Department of Pathophysiology, Hebei Medical UniversityMATERIALS:The experiment was conducted from January to April 2004at Department of Pathophysiology, Hebei Medical University. Total 40 SD rats, 7-day old, were at random divided as normal control group, model group, humgqi low-dose group and huangqi high-dose group, with 10 rats in each group. Huangqi injection (The content in 10 mL injection is consistent with 20 g raw drug) was provided by Institute of Traditional Chinese Medicine of Hebei Medical University (produced in Chengdu Di'ou Jiuhong Pharmaceutical Factory, Batch No. 0005028).METHODS:Except rats in normal group, those in the rest groups, under conscious and local anesthesia, were all given common carotid artery ligation, establishing cerebral injury model due to ischemia and anoxia. Rats in normal group were intraperitoneally injected 0.1 mL normal saline; rats in model group were intraperitoneally injected 9 g/L normal saline, 0.1 mL each day; rats in huangqi low-dose group and huangqi high-dose group were respectively given 0.1mL, 0.5 mL huangqi injection, once a day, intraperitoneally. Cerebral blood flow was detected immediately, 2 and 4days after injection. Then the rats were decapitated for collecting the brains to measure the water content in brain, the contents of NO and MDA.MAIN OUTCOME MEASURES: [1] Water contents in brains of rats in every group. [2] Cerebral blood flow, and the contents of NO and MDA.RESULTS:Totally 40 rats were involved in the trial and all entered in the final result analysis. [1] The water content in brain of each group: Compared with normal group, the content in model group was increased immediately after model establishment [(87.316±0.275)%, (88.259±0.297)% ,P ＜ 0.05 ],and did not return to the normal level at the second day [(86.973±0.265)%,(88.173±0.445)%,P ＜ 0.05]; compared with model group, the content in huangqi high-dose group was obviously decreased at second day[(88.173±0.445)%, (86.542±0.141)% ,P ＜ 0.05]. [2] Measurement of cerebral blood flow: compared with control group, the blood flow in model group was obviously decreased immediately after model establishment[(231.88±13.33), (139.54±10.58)mV,P＜ 0.05], and did not return to normal level till the 4th day [(234.57±14.38), (145.38±13.33)mV,P ＜ 0.05];compared with model group, the blood flow in huangqi low-dose group and huangqi high-dose group, at day 4, was obviously increased [(145.38±13.33),(288.45±12.89), (313.82±21.74)mV,P ＜ 0.01]. [3] The contents of NO and MDA: The contents in model group, immediately after model establishment, were obviously higher than those in normal control group [(26.55±5.23 ), ( 19.67±7.17 )μmol/L,P ＜ 0.05; (7.88±2.55), (4.22±0.12) μmol/L, P＜ 0.01], and at day 4, were significantly higher than those in normal control group [(48.65±17.06), (18.65±2.12)μmol/L,P ＜ 0.01; (5.29±0.68),(4.06±0.39)μmol/L,P ＜ 0.05]; compared with model group, the contents in huangqi low-dose group and huangqi high-dose group were obviously decreased at day 4 [(48.65±17.06), (23.77±12.79), (24.67±11.54)μ mol/L,P＜ 0.01; (5.29±0.68), (4.51±2.30), (3.68±0.39)μmol/L,P ＜ 0.01].CONCLUSION:Huangqi could obviously reduce cerebral edema from ischemia and anoxia, increase cerebral blood flow. It could decrease the contents of NO and MDA that is metabolite of free radical injury, thus playing its role to inhibit lipid peroxidation injury.

Adolescent ; Apolipoproteins E ; blood ; Humans ; Hyperlipoproteinemias ; complications ; Hypolipidemic Agents ; therapeutic use ; Kidney Diseases ; blood ; drug therapy ; pathology ; Kidney Glomerulus ; blood supply ; pathology ; Male ; Thrombosis ; pathology

Aneurysm, False ; etiology ; surgery ; Blood Loss, Surgical ; Carcinoma ; Carotid Arteries ; Carotid Artery Diseases ; etiology ; surgery ; Humans ; Nasopharyngeal Neoplasms ; radiotherapy ; Rupture

Background: During cup implantation,vertical height of the cup center(V-HCC)should be precisely controlled to achieve sufficient bone-cup coverage(BCC).Our study aimed to investigate the acetabular bone stock and the quantitative relationship between V-HCC and BCC in Crowe types Ⅰ to Ⅲ hips.Methods: From November 2013 to March 2016,pelvic models of 51 patients(61 hips)with hip dysplasia were retrospectively reconstructed using a computer software.Acetabular height and doom thickness were measured on the mid-acetabular coronal cross section.V-HCC was defined as the vertical distance of cup rotational center to the interteardrop line(ITL).In the cup implantation simulation,the cup was placed at the initial preset position,with a V-HCC of 15 mm,and moved proximally by 3-mm increments.At each level,the BCC was automatically calculated by computer.Analysis of variance and Kruskal-Wallis test were used to compare the differences between groups.Results: There were no significant between-group differences in maximum thickness of the acetabular doom; however,peak bone stock values were obtained at heights of 41.63±5.14 mm(Crowe type Ⅰ),47.58 ± 4.10 mm(Crowe type Ⅱ),and 55.78 ± 3.64 mm(Crowe type Ⅲ)above the ITL.At 15 mm of V-HCC,median BCC was 78％(75-83％)(Crowe type Ⅰ),74％(66-71％)(Crowe typeⅡ),and 61％(57-68％)(Crowe type Ⅲ).To achieve 80％of the BCC,the median V-HCC was 16.27(15.00-16.93)mm,18.19(15.01-21.53)mm,and 24.13(21.02-28.70)mm for Crowe types Ⅰ,Ⅱ,and Ⅲ hips,respectively.Conclusion: During acetabular reconstruction,slightly superior placement with V-HCC ＜25 mm retained sufficient bone coverage in Crowe Ⅰ to Ⅲ hips.

Ticks are obligate ectoparasites and vectors of arboviruses, vickettsiate, spirochetes and parasitil protozoa of humans and domestic animals. Immunological protection of mammalian hosts against tick infestation has been proposed as the most sustainable alternative tick control method to the current use of acaricides. The success of this method is dependent on the identification of key molecules for use as tick vaccine antigens. Proteolytic enzymes are involved in a wide range of cellular processes, thus they can be considered as good target antigens for a tick vaccine. In the present study, we used rapid amplification of cDNA ends protocol and primers that were designed based on the consensus amino acid motifs flanking present in all papain-like cysteine proteinases, to amplify, sequence and characterize two Rhipicephalus haemaphysaloides haemaphysaloides cathepsin L-like cysteine proteinases, named as cysA and cysB. The full length of cysA is 1168bp, encoding a 332 amino acid residue polypeptide with 36.33kD predicted molecular mass; the full length of cysB is 1153bp, encoding a 335 amino acid residue polypeptide with 37.56kD predicted molecular mass. The consensus amino acid motifs flanking presence in both deduced amino acid sequences. And both genes show high sequence homology to other tick cathepsin L-like cysteine proteinase, so they were identified as members of the cysteine proteinase gene family. Expression analysis by RT-PCR revealed that cysA and cysB were expressed differently in different periods of tick development.
Amino Acid Sequence ; Animals ; Cathepsin L ; Cathepsins ; genetics ; Cloning, Molecular ; Cysteine Endopeptidases ; genetics ; Female ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction ; Rhipicephalus ; enzymology ; Sequence Analysis

OBJECTIVETo investigate the correlation of serum sex hormones and parathyroid hormone (PTH) with the biochemical markers of bone turnover in aged men.

METHODSWe collected the laboratory data of 465 men aged 60- 93 (73. 1 +/- 8. 3) years old, who came for routine physical examinations in our hospital. We obtained the levels of serum follicle- stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), testosterone (T), sex hormone binding globulin (SHBG), PTH, 25-hydroxy-vitamin D3 (25(OH) D3), and bone turnover markers C-terminal telopeptide of type I collagen (CTX), osteocalcin (OC) and amino-terminal propeptide of type I procollagen (PINP). We also determined free testosterone (FT) , bioactive testosterone (BT) , testosterone secretion index (TSI) and FT index (FTI), and analyzed the correlation of each index with the biochemical markers of bone turnover.

RESULTSThe concentrations of serum FSH, LH, and SHBG increased, while the levels of FT, BT, TSI, FTI, PTH, CTX, OC and PINP decreased with age, especially in those over 80 years old (P <0.05). PTH was positively correlated with CTX, OC and PINP (r =0. 227, 0. 269 and 0. 162, P <0. 01), even after the adjustment for age, while SHBG negatively correlated with OC (r = -0. 100, P <0.05). The bone turnover markers increased with the elevation of the PTH quartiles, with significant differences between the first and the fourth quartile (P <0. 01). Multiple stepwise regression analysis showed that age was correlated inversely with CTX, OC and PINP ( beta = -0. 126, -0. 141 and -0. 122, P <0.05) , PTH positively with the three markers (beta = 0. 196, 0.279 and 0.189; P <0. 001), and SHBG negatively with OC ( beta = -0. 100, P <0.05) .

CONCLUSIONAging is the fundamental cause of reduced bone turnover in aged men. The levels serum PTH and SHBG are significantly associated with the biochemical markers of bone turnover.

Aged ; Aged, 80 and over ; Aging ; Bone Density ; Bone Remodeling ; physiology ; Bone and Bones ; metabolism ; Estradiol ; blood ; Follicle Stimulating Hormone ; blood ; Gonadal Steroid Hormones ; blood ; Humans ; Male ; Middle Aged ; Parathyroid Hormone ; blood ; Testosterone ; blood

OBJECTIVETo identify potential mutation of apolipoprotein E (apoE) gene in a male patient affected with lipoprotein glomerulopathy (LPG), his mother and his sister.

METHODSThe patient and his mother both had histologically confirmed LPG. His sister and his father were asymptomatic. Genomic DNA was extracted from peripheral blood samples. PCR products of the coding region of exons 3 and 4 of the apoE gene were cloned into a pTA2 vector and sequenced. Genetic variations of the apoE gene were detected using PCR and restriction fragment length polymorphism (RFLP).

RESULTSAn apoE gene mutation was identified in the patient's family. Sequence analysis confirmed a 9-bp deletion in the exon 4 of apoE gene from nt 484 to 492. The 9-bp deletion resulted in loss of 3 amino acids at positions 143-145. The sister of the propositus carried the same mutation, though she had neither proteinuria nor elevated plasma apoE. Sequence analysis of exon 3 showed no abnormality. No abnormalities were found in the father's apoE gene sequence. Analysis of genetic variations of the apoE gene by PCR and RFLP confirmed a 57 bp fragment consistent with the 9-bp deletion in exon 4. The father had a normal ε 3 ε 3 genotype.

CONCLUSIONThe 9 bp deletion of apoE may be associated with the pathogenesis of LPG.

Adolescent ; Apolipoproteins E ; blood ; genetics ; Exons ; Female ; Genetic Predisposition to Disease ; Genetic Variation ; Humans ; Kidney Diseases ; blood ; genetics ; Kidney Glomerulus ; metabolism ; pathology ; Lipoproteins ; blood ; Male ; Mutation ; Pedigree

OBJECTIVETo investigate the effects of ghrelin on the proliferation and differentiation of 3T3-L1 preadipocyte, and study the possible mechanisms.

METHODS3T3-L1 preadipocytes were cultured in vitro. The proliferation potentials of 3T3-L1 preadipocytes that were treated with different concentrations of ghrelin were evaluated by MTT methods. The levels of c-myc and thymidine kinase mRNA were detected using RT-PCR. 3T3-L1 preadipocytes were differentiated into the matured adipocytes with insulin (INS) or ghrelin. The morphological changes of 3T3-L1 adipocytes were observed and the differentiation rate was assayed by oil-red O staining. Total RNA was extracted from adipocytes at various times, and the levels of peroxisome proliferation activated receptor gamma (PPARgamma) and CAAT/enhancer binding protein(C/EBPalpha) mRNA expressions were detected using RT-PCR.

RESULTSGhrelin at concentrations of 10(-7) to 10(-15) mol/L significantly stimulated preadipocyte proliferation (p<0.05). The levels of c-myc and thymidine kinase mRNA significantly increased in 3T3-L1 preadipocytes with 10(-9) mol/L and 10(-11) mol/L ghrelin treatment (p<0.01). The 3T3-L1 preadipocytes treated with 10(-11) mol/L ghrelin had lots of lipid droplets in the cytoplasma, but the differentiation rate was lower than those treated with INS. Ghrelin of 10(-11) mol/L significantly increased the mRNA expression of PPARgamma and C/EBPalpha in the course of 3T3-L1 preadipocyte differentiation, compared with the normal control group (p<0.05). The PPARgamma and C/EBPalpha mRNA expression increased with the prolonged differentiation of preadipocytes induced by ghrelin or INS. There were significant differences in the levels of PPARgamma and C/EBPalpha mRNA expression between the 2nd and 8th days of differentiation(p<0.01).

CONCLUSIONSGhrelin promotes the proliferation and differentiation of 3T3-L1 preadipocytes. The proliferation of 3T3-L1 preadipocytes induced by ghrelin may be associated with increased c-myc levels. Ghrelin may promote differentiation of 3T3-L1 preadipocytes by increasing mRNA expression of PPARgamma and C/EBPalpha, thus enhances the sensitivity of adipocytes to INS.

3T3-L1 Cells ; Adipocytes ; cytology ; drug effects ; Animals ; CCAAT-Enhancer-Binding Protein-alpha ; genetics ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Genes, myc ; Ghrelin ; pharmacology ; Mice ; PPAR gamma ; genetics ; RNA, Messenger ; analysis ; Stem Cells ; cytology ; drug effects ; Thymidine Kinase ; genetics

OBJECTIVETo develop the characteristic chromatographic profile of Sarcandra glabra by HPLC for its quality control.

METHODThe HPLC analysis was performed on an Agilent Zorbax Eclipse XDB-C18 column (4. 6 mm x 250 mm, 5 microm) with column temperature at 40 degree C. The mobile phase was consisted of water containing 0. 5% formic acid and acetonitrile to methanol (1:9) in gradient mode, and the detection wavelength was set at 344 nm.

RESULTA common mode of the HPLC characteristic chromatographic profile has been establised. There were 20 common peaks , seven of which were identified, and 46 samples from different habitats were classified into five groups based on principal component cluster analysis.

CONCLUSIONThe method was time-saving and can represent the chemical information and provide a scientific basis for quality control of S. glabra.

Chromatography, High Pressure Liquid ; Cluster Analysis ; Drugs, Chinese Herbal ; chemistry ; standards ; Ferns ; chemistry ; Organic Chemicals ; analysis ; chemistry ; isolation & purification ; Quality Control