AIMTo develop simple but reliable intracellular labelling method for high-resolution visualization of the fine structure of single neurons in brain slice with thickness of 500 microm.

METHODSBiocytin was introduced into neurons in 500 microm-thickness brain slices while blind whole cell recording. Following processed for histochemistry using the avidin-biotin-complex method, stained slices were mounted in glycerol on special glass slides. Labelled cells were digital photomicrographed every 30 microm and reconstructed with Adobe Photoshop software.

RESULTSAfter histochemistry, limited background staining was produced. The resolution was so high that fine structure, including branching, termination of individual axons and even spines of neurons could be identified in exquisite detail with optic microscope. With the help of software, the neurons of interest could be reconstructed from a stack of photomicrographs.

CONCLUSIONThe modified method provides an easy and reliable approach to revealing the detailed morphological properties of single neurons in 500 microm-thickness brain slice. Without requisition of special equipment, it is suited to be broadly applied.

Animals ; Image Processing, Computer-Assisted ; Neurons ; cytology ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley ; Software ; Staining and Labeling ; methods

OBJECTIVETo investigate the association of peripheral and central blood pressure with the alpha-adducin Gly460Trp polymorphism in Chinese.

METHODSWe randomly selected 6 villages from JingNing County, ZheJiang Province. We invited nuclear families to take part in our study. We measured each participant's blood pressure at the non-dominant arm by means of a standard mercury sphygmomanometer at subjects' homes. Five consecutive readings were averaged for analysis. Central blood pressures were obtained by use of SphigmoCor pulse wave analysis system. The observers administered a standardized questionnaire to collect information on smoking habits, alcohol consumption and use of antihypertensive drugs. Venous blood was sampled and the adducin genotype was determined by restrictive fragment length polymorphism (RFLP).

RESULTSFour hundred and forty-two subjects included 230 (52.0%) women, and 116 (26.2%) hypertensive patients, of whom 49 (11.1%) took antihypertensive drugs. The frequencies of alpha -adducin GlyGly, GlyTrp and TrpTrp genotypes were 21.3%, 54.5% and 24.2%, respectively. There was no association between the alpha-adducin Gly460Trp polymorphism and peripheral systolic and diastolic blood pressure and pulse pressure. However, both before and after adjustment for sex, age, age(2), body-mass index, current smoking, alcohol intake, and antihypertensive treatment, the alpha-adducin polymorphism was significantly (P < 0.02) associated with central systolic blood pressure and central pulse pressure. After adjustment, central systolic blood pressure (+/- SE) averaged 122.5 +/- 3.5, 114.1 +/- 1.5 and 109.1 +/- 1.8 mm Hg (P = 0.01) in the GlyGly, GlyTrp and TrpTrp subjects, respectively. The corresponding values for central pulse pressure were 39.4 +/- 1.3, 36.4 +/- 1.0 and 32.9 +/- 0.9 mm Hg (P = 0.002), respectively.

CONCLUSIONSIn the JingNing population, the adducin 460Trp allele was associated with lower levels of central systolic pressure and pulse pressure.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Blood Pressure ; Calmodulin-Binding Proteins ; genetics ; Child ; China ; epidemiology ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertension ; epidemiology ; genetics ; physiopathology ; Male ; Middle Aged ; Pedigree ; Polymorphism, Single Nucleotide ; Young Adult

OBJECTIVETo provide right time points in selection of right aged animals and the normal physiological data of TX mice.

METHODS7-12 months old TX and DL mice were studied, each group contained 3 female and 3 male mice of TX or DL mice. The concentration of copper in the serum, dry tissues (liver, brain and kidney), together with copper biochemistry indexes were measured. The liver histopathology was observed under light microscopy and electron microscope.

RESULTSTransaminase increased significantly only in 10 and 11-month- old (AST(TX10) = 218.3 U/L, AST(TX11) = 197.5 U/L, AST(DL10) = 171.5 U/L, AST(DL11) = 165.0 U/L, P(10) less than 0.001, P(11) = 0.022), but the copper concentration of liver, brain and kidney was significantly increased during 7-12 month old (the average concentration of copper, Liver(TX) = (750.0 +/- 85.5) mg/kg, Brain(TX) = (39.7 +/- 2.2)mg/kg, Kidney(TX) = (29.8 +/- 5.0) mg/kg, Liver(DL) = (11.6 +/- 1.5) mg/kg, Brain(DL) = (16.8 +/- 0.9) mg/kg, Kidney(DL) = (14.2 +/- 1.0) mg/kg, t = 21.16, 23.60, 7.47, for all these organs P less than 0.05).

CONCLUSIONTX mice is a suitable model of liver disease with natural recovery, so selecting animal model of suitable time point is very important.

Animals ; Aspartate Aminotransferases ; blood ; Brain ; metabolism ; Ceruloplasmin ; metabolism ; Copper ; metabolism ; Disease Models, Animal ; Female ; Kidney ; metabolism ; Liver ; metabolism ; pathology ; Liver Diseases ; blood ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred Strains ; Time Factors

Twenty Newcastle disease virus (NDV) strains were isolated from chickens and geese in the field outbreaks during 2005 and 2006 in some regions of Jiangsu and Guangxi province. Assessment of the virulence by MDT and ICPI, RT-PCR and sequence analysis of fusion protein gene were used to compare the properties of NDV isolates. The results indicated that MDT and ICPI of the isolates were 45.3h - 58.2h and 1.61 - 2.00 respectively, which confirmed that the all NDV isolates were highly virulent. And their hemagglutinin were not resistant to heat and belonged to fast pattern of elution. The results of nucleotide sequencing and phylogentic analysis of fusion protein gene showed that the twenty strains shared homology from 79.7% to 100% among themselves, from 78.1% to 83.4% and from 80.2% to 90.1% with NDV LaSota, F48E8, respectively. The putative amino acid sequences of fusion protein at the cleavage sites of all the isolates were 112R-R-Q-R/K-R-F117, with the motif characteristics of the virulent NDV strain, which was in accordant with the results of assessment of the pathogenicity. The phylogentic tree based on sequences of fusion protein gene variable regions (47-420nt) revealed that the 18 strains belonged to sub-genotype VIId and the others belonged to an old genotype III of NDV, revealing that subgenotype VIId virus was responsible for the NDV outbreaks in some regions of Jiangsu and Guangxi promince recently.
Amino Acid Motifs ; Amino Acid Sequence ; Animals ; Base Sequence ; Chickens ; virology ; China ; epidemiology ; Disease Outbreaks ; Geese ; virology ; Molecular Epidemiology ; Newcastle Disease ; epidemiology ; genetics ; Newcastle disease virus ; genetics ; pathogenicity ; Phylogeny ; Poultry Diseases ; epidemiology ; genetics ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Fusion Proteins ; genetics

AIMTo determine the possible roles of the t-complex testis expressed gene 5 (Tctex5) on sperm functions, the full-length sequence of mRNA was studied and compared in the testis between the normal wild-type and the sterile t-haplotype mutant mice.

METHODSWe applied rapid amplification of cDNA ends, Northern blot and reverse transcription polymerase chain reaction to analyze the full length of Tctex5 mRNAs isolated from testes of the wild-type and the t-haplotype mice. Reverse transcription polymerase chain reaction was used to semi-quantitatively compare expression of Tctex5 transcripts in the 16 tissues and 9.5 day stage embryos in the wild-type mice. E-translation was applied to estimate the amino acid sequences.

RESULTSOne long and one short transcript of Tctex5 mRNA were discovered in mouse testis of wild-type (Tctex5(long-+) and Tctex5(short-+)) and t-haplotype (Tctex5(long-t) and Tctex5(short-t)) mice, respectively. Being enhanced only in the testis, Tctex5(long-t) had 17 point mutations and one 15-bp-deletion in the exon 1 region, comparing with the Tctex5(long-+), whereas the Tctex5(short-t) was similar to the Tctex5(short-+). The short isoforms of Tctex5 mRNAs in the two models encoded exactly the same peptides, but the long isoforms did not. The estimated peptide encoded by Tctex5(long-t) had significant mutations on putative sites of phosphorylation and PP1 binding.

CONCLUSIONWe established that mutations that occur in the Tctex5 long transcript of the t-haplotype mice are important for normal sperm function, whereas the short transcript of Tctex5 might have a conserved function among different tissues.

Animals ; Gene Expression ; Haplotypes ; Infertility, Male ; Male ; Mice ; Microtubule-Associated Proteins ; chemistry ; genetics ; Mutation ; Nuclear Proteins ; chemistry ; genetics ; Protein Phosphatase 1 ; Sequence Analysis, Protein ; Spermatozoa ; metabolism ; Testis ; metabolism ; t-Complex Genome Region

Hepatitis B ; prevention & control ; transmission ; Hepatitis B virus ; Humans ; Infectious Disease Transmission, Vertical ; prevention & control

BACKGROUNDIt remains almost a helpless situation for the recurrent implantation failure and pregnancy loss caused by endometrial injury at present. The purpose of this study was to develop a rabbit model of endometrial mechanical injury that could provide a research platform for this difficult clinical predicament.

METHODSThree experiments were conducted. Experiment 1: Curettages in both uterus horns and copper wire inserting after curettage (double-injury) in one horn. The histological changes were monitored at 0, 24, 48, 72 hours, as well as in 1 and 2 weeks after operation. Experiment 2: Direct copper wire inserting in one horn and double-injury in other horn. The wires in both horns were removed after 2 weeks. The histological changes were recorded at 0, 1 and 2 weeks after wire removal. Experiment 3: Double-injury procedure in one horn was performed and wire was removed after 2 weeks; another horn was remained normal to serve as control. Histological changes were recorded, tissue areas were measured, and proliferation indices (PIs, %) were calculated at 1, 2, 4 and 8 weeks after wire removal, respectively.

RESULTSThe experiments revealed that the injured endometrium by simple curettage or copper wire could be fully repaired. While the endometrial regeneration was severely impaired by double-injury, both areas of endometrium and uterine cavity decreased (P < 0.05); both PIs of glandular epithelial and stromal cells increased and reached maximum at 4 weeks (P < 0.05), but returned by 8 weeks.

CONCLUSIONThis study demonstrated that a rabbit model of endometrial injury could be effectively established through a double-injury procedure of curettage and copper wire with comparable clinical index.

Animals ; Copper ; adverse effects ; Curettage ; adverse effects ; Disease Models, Animal ; Endometrium ; injuries ; Female ; Immunohistochemistry ; Rabbits

OBJECTIVETo design and synthesize a series of squamosamide cyclic analogues and to test their antioxidation activity.

METHODSEleven 3-substituted indole-2-one derivatives were designed and synthesized through 9 steps with p-hydroxyphenylacetic acid as the starting material and their structures were confirmed by nuclear magnetic resonance and mass spectrometry.

RESULTSEleven compounds showed antioxidation activity and the activities of compounds 9 and 13 matches the positive control FLZ-52.

CONCLUSIONCyclic reconstruction with FLZ-52 as the lead compound have some antioxidation activity.

Animals ; Annonaceae ; chemistry ; Antioxidants ; chemical synthesis ; chemistry ; pharmacology ; Benzeneacetamides ; chemical synthesis ; chemistry ; pharmacology ; Phenols ; chemical synthesis ; chemistry ; pharmacology ; Rats

OBJECTIVETo study the effect of prosthetic replacement in treatment of femoral neck fractures on the hemiplegia side in the elderly.

METHODSFrom May 1990 to May 2000, 189 elderly patients with femoral neck fractures were treated with prosthetic replacement in my hospital. Twenty-nine hemiplegia patients, who suffered from stroke previously, had Garden type III and type IV femoral neck fractures on the hemiplegia side. Thirty non-hemiplegia patients were chosen randomly. The two groups were followed-up for 27-98 months (average: 59 months). The age, hospitalization days, operating time, blood loss, blood transfusion, complications during perioperative period and long-term complications were compared between the two groups and the results of femoral head replacement and total hip replacement in the hemiplegia group were also compared.

RESULTSAll the patients of the two groups survived the perioperative period. No significant difference was found in the age, hospitalization days, operation time, blood loss and blood transfusion and long-term complications between the two groups (P>0.05). However there was significant difference in complications during perioperative period between the two groups (P<0.05). Five patients died in the hemiplegia group with the mortality of 17.2% and two died in the non-hemiplegia group with the mortality of 6.7% 11 months to 5 years after operation. There was significant difference in long-term complications between the femoral head replacement and the total hip replacement in the hemiplegia group (P<0.05). The result of the total hip replacement was better than that of the femoral head replacement.

CONCLUSIONSProsthetic replacement is a reliable method in treatment of Garden type III and type IV femoral neck fractures on the hemiplegia side in the elderly, and patients are safe during perioperative period. More complications during perioperative period occur in the hemiplegia group, and long-term complications are insignificantly different between the two groups. The mortality rate is higher in the hemiplegia group than in the non-hemiplegia group within 5 years after operation. Since the result of the total hip replacement is better than that of the femoral head replacement, total hip replacement should be chosen firstly to treat Garden type III and type IV femoral neck fractures on the hemiplegia side in the elderly if the muscular strength of the hip is beyond IV degree.

Aged ; Arthroplasty, Replacement, Hip ; Female ; Femoral Neck Fractures ; etiology ; surgery ; Hemiplegia ; complications ; Humans ; Male

Background: and Purpose This study aimed to construct an optimal dynamic nomogram for predicting malignant brain edema (MBE) in acute ischemic stroke (AIS) patients after endovascular thrombectomy (ET). Methods: We enrolled AIS patients after ET from May 2017 to April 2021. MBE was defined as a midline shift of >5 mm at the septum pellucidum or pineal gland based on follow-up computed tomography within 5 days after ET. Multivariate logistic regression and LASSO (least absolute shrinkage and selection operator) regression were used to construct the nomogram. The area under the receiver operating characteristic curve (AUC) and decisioncurve analysis were used to compare our nomogram with two previous risk models for predicting brain edema after ET. Results: MBE developed in 72 (21.9%) of the 329 eligible patients. Our dynamic web-based nomogram (https://successful.shinyapps.io/DynNomapp/) consisted of five parameters: basal cistern effacement, postoperative National Institutes of Health Stroke Scale (NIHSS) score, brain atrophy, hypoattenuation area, and stroke etiology. The nomogram showed good discrimination ability, with a C-index (Harrell’s concordance index) of 0.925 (95% confidence interval=0.890–0.961), and good calibration (Hosmer-Lemeshow test, p=0.386). All variables had variance inflation factors of <1.5 and tolerances of >0.7, suggesting no significant collinearity among them. The AUC of our nomogram (0.925) was superior to those of Xiang-liang Chen and colleagues (0.843) and Ming-yang Du and colleagues (0.728). Conclusions Our web-based dynamic nomogram reliably predicted the risk of MBE in AIS patients after ET, and hence is worthy of further evaluation.