BACKGROUND: Synaptic density, a key index of structure and function of brain tissues, is related to cognitive function. Synaptic loss occurs during human brain aging and in Alzheimer disease (AD), inducing the changes of synaptic density.OBJECTIVE: To observe quantitative synaptic alterations in human brain and changes of synaptic density in different parts during normal aging so as to compare them with those of AD patients.DESIGN: Sampling survey.SETTING: Senile Neurological Department of General Hospital of Chinese PLA.PARTICIPANTS: Pathological data were selected from General Hospital of Chinese PLA from June 1996 to December 2002. Inclusion criteria: had no major nervous system diseases and neuropathological changes. Brain tissues of 28 corpses in normal aging group, 23 males and 5 females aged 23-100 years with an average of (65±22.8) years, were obtained at autopsy.All corpses were divided into three groups according to their age, namely,adult group (23-55 years old, n=9), senile group (64-72 years old, n=7),and ＞75 group (76-100 years old, n=12). Cerebral hippocampal samples of other six corpses diagnosed with AD were selected from clinic. The corpses included 5 men and 1 woman aged 76-94 years with an average of (83±7.7) years.METHODS: Response intensity of synaptophysin immunochemistry remained stable after 4-8 hours of death, so brains were obtained at autopsy after 8-72 hours of death and fixed with 4% formalin for at least 6 weeks.In normal aging group, tissues were taken from left superior frontal gyrus,striatal area of left occipital lobe, left putamen (striatum section, including head of caudate nucleus), and left hippocampus (from lateral geniculate body section to medial occipitotemporal gyrus). In AD cases, tissues were taken from left hippocampus of 4 corpses and right hippocampus of other 2. All sections were stained with hematoxylin eosin (HE), toluidine blue and synaptophysin immunostaining (rabbit anti-human synaptophysin polyclonal antibody from Beijing Zhongshan Biotechnology Co., Ltd.). Morphology and distribution of positive objects in synapse immunologic reaction were observed under the light microscope. Relation between absorbance in each region and age was determined with Pearson's coefficient. Differences among groups were analyzed with nonparametric test, and the differences in hippocampal CA3 area between ＞ 75 group and AD group were analyzed with the same test.MAIN OUTCOME MEASURES:① Absorbency of synaptophysin at various sites of normal aging group and correlation with age; ② absorbance value in CA3 area between AD patients and advanced aged normal subjects (＞75 years) was compared.RESULTS:All the 34 cerebral samples entered the final analysis.①Synaptophysin-positive granules of various size were scattered through neocortex, putamen and hippocampus, neuronal somata, neuroglia, vessels and white matter. Density was particularly strong over layers Ⅱ and Ⅲ in frontal lobe, and layer ⅣV in occipital lobe. ② Synaptophysin density was negatively correlated with age, which was -0.688 in frontal lobe, -0.592 in occipital lobe, -0.458 in putamen and -0.619 in hippocampal CA2 area,respectively (P = 0.000, 0.001, 0.014, and 0.000). ③ Significant difference in synaptic density in CA3 area was found between AD patients (0.031 3±0.003 0)and normal subjects over the age of 75 (0.040 7±0.005 3) (Z=-2.997, P=0.001)in nonparametric test.CONCLUSION:① Synaptic density was found to decrease in frontal lobe, occipital lobe, CA3 area of hippocampus and putamen with age; the changes had significant correlation with age.② Synaptic density of AD patients was lower than that of normal subjects, and their cognitive hypofunction was related to synaptic loss. ③ All tissues were obtained after 8-72 hours of death and fixed over 6 weeks, which to the greatest extent reduced the effects of tissue autolysis and formalin fixation on the results.

Human-animal parasitic diseases caused by medical helminths are hazardous to human health. Genetic polymorphism studies on medical helminth populations can not only understand the biological characteristics and genetic structure of their populations, but also help reveal how they adapt to their parasitic environment, thus contributing to deepen our understanding of the epidemiological patterns of parasitic diseases and improve our understanding of accurate prevention and control of parasitic diseases. With the development of molecular biology, molecular markers such as DNA barcodes, simple sequence repeats, and single nucleotide polymorphism markers have been widely used to study the genetic relationships among parasite populations and individuals, and to reveal the genetic variation of parasite populations and the evolution of species origins. In this paper, we systematically review the application of three molecular markers commonly used in the study of genetic polymorphism in medical helminths, with a view to laying the foundation for related research.

OBJECTIVETo investigate morphological changes of capillary in aging brain and explore the role of vascular factor in brain aging.

METHODSTwenty-eight brains of individuals (mean age 65 years) who died without clinical or pathological involvement of nervous system and 6 brains of Alzheimer's disease (AD) patients (mean age 83 years) were obtained at autopsy. Sections from frontal lobe, occipital lobe, striatum and hippocampus of normal subjects and sections from hippocampus of AD patients were used for hematoxylin eosin (HE), lox fast blue (LFB), toluidine blue stains and ulex europaeus agglutinin (UEA) immunostaining. After observations of morphological changes of neuron and capillary, computer-aid image analysis was performed to quantify numerical density and area density of neuron and capillary in frontal lobe, occipital lobe, putamen, CA3 sector of normal subjects and CA3 sector of AD patients. Numerical ratio and area ratio of neuron and capillary were then calculated. Correlations between neuron/capillary ratio and age were estimated using Pearson's correlation test. Difference of neuron/capillary ratio in CA3 sectors between AD patients and advanced aged normal subjects (> 75 years) was analyzed with Student's t-test.

RESULTSSeveral pathological microvascular changes, including increased tortuosity, looping, bundling, stringing, and effacement of endothelia were seen in aged subjects and more prevalent in AD patients. Numerical ratio and area ratio of neuron and capillary of frontal lobe, occipital lobe and putamen significantly increased with age in normal aging subjects.

CONCLUSIONSMorphological changes and relative decrease in number and capacity of capillary in aging brain may reduce cerebral blood flow and metabolism, and consequently result in functional impairment of aging brain. Vascular factors may play an important role in the development of brain aging.

Adult ; Aged ; Aged, 80 and over ; Aging ; Alzheimer Disease ; etiology ; pathology ; Capillaries ; anatomy & histology ; pathology ; Cell Count ; Cerebral Cortex ; blood supply ; pathology ; Female ; Frontal Lobe ; blood supply ; pathology ; Hippocampus ; blood supply ; pathology ; Humans ; Image Processing, Computer-Assisted ; Male ; Middle Aged ; Neurons ; pathology ; Occipital Lobe ; blood supply ; pathology

OBJECTIVETo investigate telomerase activity in rabbit bone marrow stromal cells (BMSCs) during their committed differentiation in vitro along neural pathway and the effect of glial cell line-derived neurotrophic factor (GDNF) on the expression of telomerase.

METHODSBMSCs were acquired from rabbit marrow and divided into control group, GDNF (10 ng/ml) group. Cytokine.NSCs medium (prepared by our lab, Patent No. ZL02134314. 4) supplemented with 10 percent fetal bovine serum (FBS) was used to induce BMSCs differentiation along neural pathway. Fluorescent immunocytochemistry was employed to identify the expressions of Nestin, neuron-specific endase (NSE), and gial fibrillary acidic protein (GFAP). The growth curves of the cells and the status of cell cycles were analyzed, respectively. During the differentiation, telomerase activities were detected using the telomeric repeat amplification protocol-enzyme-linked immunosorbent assay (TRAP-ELISA).

RESULTSBMSCs were successfully induced to differentiate along neural pathway and expressed specific markers of fetal neural epithelium, mature neuron and glial cells. Telomerase activities were undetectable in BMSCs during differentiation along neural pathway. Similar changes of cell growth curves, cell cycle status and telomerase expression were observed in the two groups.

CONCLUSIONSRabbit BMSCs do not display telomerase activity during differentiation along neural pathway. GDNF shows little impact on proliferation and telomerase activity of BMSCs.

Animals ; Bone Marrow Cells ; enzymology ; Cell Differentiation ; Glial Cell Line-Derived Neurotrophic Factor ; Immunohistochemistry ; Rabbits ; Stromal Cells ; enzymology ; Telomerase ; metabolism

OBJECTIVETo analyze the clinical features of patients with newly diagnosed myeloma bone disease (MBD).

METHODClinical features of MBD in two hundred and five patients with newly diagnosed multiple myeloma (MM) were analyzed retrospectively. The relationship between outcome of different grades of MBD patients and their prognosis was compared.

RESULTS(1) Among the 205 patients, one hundred and fifty (72.7%) had bone pain as the first symptom. (2) According to X-ray evaluation, there were 23 (11.3%) of grade 0, 14 (6.9%) grade 1, 23 (11.3%) of grade 2, 68 (33.3%) grade 3 and 76 (37.2%) grade 4. (3) Patients with grade2-4 MBD had significantly higher ECOG performance score, marrow plasmacytes, marrow CD138(+) CD38(+) cell percentage and serum IL-6 level than those with grade 0 - 1 did (P < 0.05). Patients with grade 4 MBD presented with hypocalcemia (P < 0.05) more often than those with grades 0 - 3 diseases did. (4) There was no significant difference in response to initial induction chemotherapy among the five groups (P = 0.642). (5) Univariate analysis demonstrated that the time to progression (TTP) in grade 2 - 4 MBD groups was significantly shorter than that in grade 0 - 1 groups (P = 0.029). (6) Multivariable COX analysis did not indicate lytic bone changes was a independent prognostic factor for OS and TTP.

CONCLUSIONThere is a rather high incidence of MBD in MM patients in China. Patients with extensive X-ray bone lesions have more severe hematologic parameters than those without bone lesions did, and severe bone lesions is an important adverse prognostic factor for TTP.

Bone Diseases ; Bone Marrow ; Humans ; Interleukin-6 ; Multiple Myeloma ; Prognosis ; Retrospective Studies

OBJECTIVETo explore a cosmetic incision in the neck dissection for differentiated thyroid carcinoma.

METHODSAn extended collar incision was used for neck dissection in 82 consecutive patients with thyroid carcinoma from May 1999 to December 2006. The incision was designed to start as a conventional thyroid collar incision, and then to extend it along the skin crease to the anterior border of trapezium, so to avoid the vertical limb of conventional hockey stick incision. There were 60 females and 22 males in this series, with a median age of 40.5 years (range, 10 to 80 years). Ninety-six procedures of neck dissection were performed in 82 patients, including one radical neck dissection, one type I modified neck dissection, 8 type II modified neck dissections, and 86 type III modified neck dissections.

RESULTSThe average time of anesthesia was 197 minutes. The average dissected lymph nodes were 37.5, with average metastasis in 8.8 nodes. Eight patients (9.8%) developed complications related to neck dissection. The follow-up period in these patients were 1 to 96 months with a median follow-up time of 23 months. Cervical recurrence was found in only one patient (1.2%). Neither death nor distant metastasis was observed in this series.

CONCLUSIONIt is feasible to perform a modified neck dissection for differentiated thyroid cancer through the extended collar incision. The preliminary results show that the above described incision is not only oncologically safe, but also offers a cosmetic benefit for the patient with thyroid carcinoma.

Adenocarcinoma, Follicular ; pathology ; surgery ; Adenocarcinoma, Papillary ; pathology ; surgery ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Follow-Up Studies ; Humans ; Hypocalcemia ; etiology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neck Dissection ; adverse effects ; methods ; Thyroid Neoplasms ; pathology ; surgery ; Young Adult

Objective To observe the effects of acupuncture at "Sanyinjiao" (SP6) and "Baihui" (GV20) on behaviors and contents of monoamine neurotransmitter dopamine (DA), noradrenalin (NE) and 5-hydroxytrypamine (5-HT) in prefrontal cortex (PFC) of attention deficit hyperactivity disorder (ADHD) rats; To discuss mechanism of action of acupuncture for prevention and treatment of ADHD. Methods 4-week-old SHR rats were randomly divided into the model group, Western medicine group and acupuncture group, and WKY rats with the same age were selected as the normal control group, with 10 rats in each group. Acupuncture was applied to "Sanyinjiao" (SP6) and "Baihui" (GV20) of rats in acupuncture group for 15 min. The rats in Western medicine group was given MPH (ritalin) for gavage, and rats were treated for 4 weeks in the dark-phase. Open field test (OFT), elevated plus maze (EPM) and novel object recognition test (NORT) were conducted to evaluate the spontaneous activity, impulsivity and learning-memorial ability of rats individually at the end of treatment. Contents of DA, NE and 5-HT in PFC were detected by HPLC. Results Compared with the normal control group, all of the behavior parameters of rats in model group were significantly higher (P＜0.05), and the contents of DA, NE and 5-HT in PFC of model group were lower than those of normal control group (P＜0.05). Compared with the model group, acupuncture group showed shorter movement distance, less rearing and fewer grooming activities in the OFT; percentages of times of entering the open arms and staying duration were reduced in the total period in the EPM; preference index in the NORT was elevated (P＜0.05); the contents of NE and 5-HT in PFC of acupuncture group increased significantly (P＜0.05). Conclusion Acupuncture at "Sanyinjiao" (SP6) and "Baihui" (GV20) canreduce the spontaneous activity and impulsivity, and improve learning and memory ability of ADHD model rats, which may be related to elevated contents of NE and 5-HT in the PFC.

OBJECTIVETo evaluate the role of parathyroid hormone (PTH) and serum calcium in prediction for hypocalcaemia after total thyroidectomy.

METHODSOne hundred and sixty-five patients undergoing total or complete total thyroidectomy were reviewed retrospectively. The indications included bilateral carcinoma, undifferential carcinoma, surroundings invasion, distant metastasis and huge benign lesions. Preoperative and postoperative PTH, calcium concentrations and their decline levels were compared between Jan. 2005 and May 2009. The role of PTH value and decline level predicting for symptomatic hypocalcaemia were analyzed by receiver operator characteristics (ROC) curve.

RESULTSAfter total thyroidectomy, 85 patients (51.5%) developed hypocalcemia. Symptoms were reported by 36 patients (21.8%). The mean concentration of PTH for normocalcaemia (80 cases), asymptomatic hypocalcaemia (49 cases) and symptomatic patients (36 cases) were 31.0 ng/L, 19.6 ng/L and 11.9 ng/L, respectively. The mean decline level for the three groups were 28.6%, 52.6% and 78.0%, respectively. PTH value and its decline level had a poor predicting value for symptomatic hypocalcaemia and high negative predicting value for asymptomatic patients. The serum calcium concentration more than 2.0 mmol/L, PTH level higher than 15 ng/L and PTH decline less than 50% had the good negative predicting value of 97.6%, 90.3% and 96.5%, respectively.

CONCLUSIONSPostoperative PTH and its decline level were significantly correlated with postoperative serum calcium concentration but had a low accuracy for predicting symptomatic hypocalcaemia. The serum calcium concentration more than 2.0 mmol/L, PTH level higher than 15 ng/L and PTH decline less than 50% had the good predicting value for asymptomatic patients. Calcium should be routinely supplemented in the first 24 h after total thyroidectomy to reduce the rate of hypocalcemia and the severity of hypocalcemia symptoms.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Calcium ; blood ; Child ; Female ; Humans ; Hypocalcemia ; blood ; diagnosis ; Male ; Middle Aged ; Parathyroid Hormone ; analysis ; Postoperative Period ; Retrospective Studies ; Thyroid Neoplasms ; surgery ; Thyroidectomy ; adverse effects ; Young Adult

ObjectiveThe genotypes and drug-resistance mutation of hepatitis B virus (HBV) are closely related to the progression of chronic hepatitis B (CHB) and effects of medication. This study was to identify the genotypes and drug-resistance mutation genes of HBV in West Guangxi and provide some laboratory evidence for anti-HBV treatment.MethodsWe collected serum samples from 869 CHB patients with HBV DNA >1.0×103 IU/mL, all from West Guangxi, between January 2016 and March 2018. We detected the genotypes and drug-resistance mutation genes in the patients by PCR and reverse dot blot (PCR-RDB).ResultsA total of 7 genotypes were identified in the patients, including genotypes B in 304 cases (34.98%), C in 268 (30.84%), D in 147 (16.92%), B+C in 28 (3.22%), B+D in 22 (2.53%), C+D in 1 (0.12%) and B+C+D in 1 (0.12%). Drug-resistance mutation was observed in 63 cases (7.25%), including 31 cases with genotype C (49.21%) and 19 with genotype B (30.16%). Fourteen gene mutation patterns were found in all, including rt204I, rt181V, rt236T, rt180M+rt204V, and rt180M+rt204V+rt204I, among which, rt180M+rt204V exhibited the highest mutation frequency (17.46%). There was a statistically significant difference between the mutation rates of genotypes B and C (19% vs 31%, P < 0.05).ConclusionCHB patients in West Guangxi have a wide variety of genotypes of HBV, mainly including genotypes B and C, with a high rate and complicated patterns of drug-resistance mutation, which has provided some reference for anti-HBV treatment in West Guangxi.

The aim of this study was to investigate the support effects of mesenchymal stem cells (MSCs) on umbilical cord blood (UCB) CD34+ cell (HSPC) expansion in vitro and its influence on cell characteristics including the surface marker of CD34+ cells, homing adhesion molecules and colony-forming ability. The mononucleated cells (MNCs) were isolated from UCB, then the CD34+ cells were isolated from freshly obtained MNCs by immunomagnetic beads, the MSC feeder cells exposed to gamma-ray of 137Cs were prepared by MSC feeder. The CD34+ cells were inoculated in different culture media. Experiment was divided into 3 groups: HSPC+CK group in which cytokines were added to medium (SCF, FL and TPO); HSPC+MSC group in which CD34+ cells were inoculated on MSC feeder; HSPC+MSC+CK group in which cytokines and MSC feeder cells were added to medium. After culture for 4, 7, 10, 14 days the MNC amount was counted and expansion ability of CD34+ cells was evaluated. The immunotypes of CD34+ cells and subsets, homing adhesion molecules and colony-forming ability in different groups detected by flow cytometry. The results showed that the amount of MNCs and CD34+ cells all obviously increased during culture for 14 days, the expansion levels of MNCs in 3 groups were HSPC+MSC+CK group>HSPC+CK group>HSPC+MSC group in proper order. Within 10 days of expansion in vitro amount of MNCs obtained significant expansion, meantime the expansion of CD34+ cells was higher also. The CD34+ count in 3 groups at day 4 of culture decreased significantly as compared with 0 day of culture (p<0.01). The CD34+ cells ratios in 3 groups after expansion were HSPC+MSC group>HSPC+MSC+CK group>HSPC+CK group in proper order (p<0.01), while CD34+ subset levels in 3 groups were different, the CD34+CD38- cells in HSPC+CK group at 4 days of culture increased transiently (62.71%), then quickly decreased, the CD34+CD38- cell ratio at day 7 was 0.05%, while the CD34+CD38- cell ratio in HSPC+MSC group at day 7 was 18.92%, difference was significant as compared with HSPC+CK group (p<0.05). The analysis of colony-forming units showed that the colony-forming ability at various time points after expansion all sustained in high level. It is concluded that in short-time (<7 day) culture of UCB CD34+ cells the combination of MSCs with cytokines can significantly expand the CD34+ cells and make the HSPCs to maintain original biologic characteristics.
Antigens, CD34 ; Cell Culture Techniques ; Cell Proliferation ; Cells, Cultured ; Culture Media, Serum-Free ; Fetal Blood ; cytology ; immunology ; Flow Cytometry ; Humans ; Immunophenotyping ; Mesenchymal Stromal Cells ; cytology ; immunology