Gut microbial communities are likely remodeled in tandem with accumulated physiological decline during aging, yet there is limited understanding of gut microbiome variation in advanced age. Here, we performed a metagenomics-based enterotype analysis in a geographically homogeneous cohort of 367 enrolled Chinese individuals between the ages of 60 and 94 years, with the goal of characterizing the gut microbiome of elderly individuals and identifying factors linked to enterotype variations. In addition to two adult-like enterotypes dominated by Bacteroides (ET-Bacteroides) and Prevotella (ET-Prevotella), we identified a novel enterotype dominated by Escherichia (ET-Escherichia), whose prevalence increased in advanced age. Our data demonstrated that age explained more of the variance in the gut microbiome than previously identified factors such as type 2 diabetes mellitus (T2DM) or diet. We characterized the distinct taxonomic and functional profiles of ET-Escherichia, and found the strongest cohesion and highest robustness of the microbial co-occurrence network in this enterotype, as well as the lowest species diversity. In addition, we carried out a series of correlation analyses and co-abundance network analyses, which showed that several factors were likely linked to the overabundance of Escherichia members, including advanced age, vegetable intake, and fruit intake. Overall, our data revealed an enterotype variation characterized by Escherichia enrichment in the elderly population. Considering the different age distribution of each enterotype, these findings provide new insights into the changes that occur in the gut microbiome with age and highlight the importance of microbiome-based stratification of elderly individuals.
Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Bacteroides ; China ; Diabetes Mellitus, Type 2/microbiology* ; Escherichia coli/classification* ; Gastrointestinal Microbiome/genetics* ; Metagenomics ; East Asian People

BACKGROUND:Mineralized nodules are the mature marker of osteoblast differentiation, and the observation methods mainly use alizarin red staining.
OBJECTIVE:To compare the observation results of mineralized nodules by three methods, and to explore their characteristics and advantages, as wel as further application in the research of bone disease.
METHODS:The rat osteoblast-like cellline UMR-106 were cultured in the fresh medium that was changed every day, for 14 days. Alizarin red staining-light microscope, tetracycline fluorescence labeling-laser confocal scanning microscopy, and scanning electron microscopy were used to observe mineralized nodules. The calcium content of mineralized nodules was quantified using scanning electron microscopy and energy dispersive spectroscopy. In addition, tumor necrosis factor alpha that could inhibit the proliferation and differentiation of osteoblasts was used as the control.
RESULTS AND CONCLUSION:The three morphology methods could be used to observe the mineralized nodules of normal osteoblasts. As for tumor necrosis factor alpha, no mineralized nodules of osteoblasts were observed by alizarin red staining-light microscopy;smal mineralized nodules were observed by tetracycline staining-laser scanning confocal microscopy and scanning electron microscopy, suggesting tetracycline staining and scanning electron microscopy were more sensitive in the observation. Scanning electron microscopy could be used to observe the submicroscopic structures of mineralized nodules in the osteoblasts, and the formation of mineralized nodules, including the calcium secretion. Additional y, scanning electron microscopy combined with energy dispersive spectroscopy analysis can successful y quantify and position the mineralized nodules, indicating a potential application in the research of bone diseases.

Objective To investigate the influence of processed total motile sperm (PTMS) count of husband on clinical pregnan-cy rate of intrauterine insemination(IUI) .Methods We retrospectively analyzed a total of 229 cycles of IUI among 131 patients in our hospital during the past three years .The cycles were divided into 5 groups according to the PTMS count :group A(<5 × 106 ) , group B(5 × 106 - <10 × 106 ) ,group C(10 × 106 - <20 × 106 ) ,group D(20 × 106 - <30 × 106 ) ,group E(≥30 × 106 ) .The clinical pregnancy rates among different groups were compared .Results The clinical pregnancy rates among 131 patients of IUI was 26 .72% .The clinical pregnancy rates among 229 cycles of IUI was 15 .28% .The clinical pregnancy rate of group A was 23 .08% , the clinical pregnancy rate of group B was 15% ,the clinical pregnancy rate of group C was 18 .18% ,the clinical pregnancy rate of group D was 13 .46% ,the clinical pregnancy rate of group E was 13 .48% .There was no significant difference(P>0 .05) among five groups .Conclusion Ideal clinical pregnancy can be achieved when the PTMS count is between 3 × 106 and 5 × 106 .

BACKGROUND:Tougu Xiaotong capsule is the clinical prescription for the treatment of osteoarthritis in Fujian University of Traditional Chinese Medicine, and the previous studies mainly focus on effect to cartilage.
OBJECTIVE:To observe the effect of Tougu Xiaotong capsule on the proliferation and differentiation of osteoblasts as wel as the expressions of bone remodeling correlated factors.
METHODS:Rat osteoblast-like cel line ROS17/2.8 cel s were incubated with Tougu Xiaotong capsule. The ROS17/2.8 cel s were divided into blank control group and Tougu Xiaotong capsule groups with different
concentrations. The cel proliferation was determined by methylthiazolyldiphenyl-tetrazolium bromide assay. Osteoblast differentiation biomarkers alkaline phosphatase activity, osteocalcin and bone mineralized nodules were measured with colorimetry, enzyme-linked immunosorbent assay and alizarin red staining, respectively. The real-time PCR and enzyme-linked immunosorbent assay were used to detect the expressions of bone remodeling factors osteoprotegerin/receptor activator of nuclear factorκB ligand.
RESULTS AND CONCLUSION:Compared with the control group, the Tougu Xiaotong capsule with the
concentration of 0.25-2 g/L could significantly promote the ROS17/2.8 cel proliferation (P<0.05), up-regulate the alkaline phosphatase activity, osteocalcin expression level and mineralized nodules area, and increase the
percentage of bone remodeling factors osteoprotegerin/receptor activator of nuclear factorκB ligand (P<0.05). The mechanism of Tougu Xiaotong capsule protecting osteoarthritis may partly result from the regulation of
proliferation and differentiation of osteoblasts and bone remodeling.

Microscopic and histochemical methods were used to investigate flavonoids localization in the leaf and the stem of the Sarcandra glabra. The results indicated that flavonoids distributed mainly in epidermis, collenchyma, vascular bundles, secretory cells and palisade tissue of leaf. In the stem, they distributed mainly in epidermis, collenchyma, phloem and secretory cells. Histochemical localization of flavonoids using 5% solution of NaOH is convenient, rapid and reliable. The content of flavonoids in the leaf was higher those than in the stem. For sustainable utilization of the resources we suggested that only the leaves could be harvested.
Flavonoids ; metabolism ; Magnoliopsida ; metabolism ; Microscopy ; Plant Leaves ; metabolism ; Plant Stems ; metabolism

Objective To study the effect of Jiedu-xiaozheng decoction on theUltrastructure changes and quantitative analysis of SGC-7901 in vitro and its mechanisms. Methods The compound prescription Jiedu-xiaozheng decoction was used to prepare serum containing drug. Serologic pharmacology methods was applied. The decoction was used to co-culture the gastric cells for24 h and 72 h, and the ultrastructure of tumor cells was observed by the transmission electron microscope. Then the parameters of the nucleus and mitochrondria were determined in SGC-7901 cells, including volume density(Vv) ,form factor PE( PE), average volume(V) ,length(L) average surrace area(S),contrasting to cytoplasma. The SIS powervision was applied to analyze their difference. Results Jiedu-xiaozheng decotion can result in reduced microvilli, deflated volume, decreased mitochondrion volume and reduced ridge. The volume density(Vv) ,form factor PE(PE) ,average volume(v) ,length (L) average surrace area(S) reduced in the 24 h group clearly in Jiedu-xiaozheng decoction group compared to control group. Conclusion Jiedu-xiaozheng decoction can significantly induce SGC-7901 cells apoptosis, exhibiting certain early ultrastructure changes such as marginated heterochromatin and mitochondrion pyknosis. The quantity analysis by electronic imaging system may objectively reveal the ultrastructural changes of cell nucleus and mitochondria.

Objective To investigate the influence of pinealectomy(Px)and melatonin (MLT) supplementation on CD4~+CD25~+Treg cells development and Foxp3 expression in rat thymus. Methods One hundred and twenty clean-grade male SD rats were divided into five groups: normal control group, sham pinealectomized group, pinealectomized group, Px +7.5 mg/(kg·d) MLT group, Px +15 mg/(kg·d) MLT group, and the thymus were taken out at the 4th week and the 8th week;Flow cytometric analysis was used to analyse the number of double positive cells in the thymus and peripheral blood;Semiquantitative RT-PCR and immunohistochemistry were applied to analyse Foxp3 expression of rat thymus. Results There was no difference of the number of CD4~+CD25~+ Treg cells in rat thymus between Px and normal/sham group, and the number increased dependently on time and dose after MLT supplementation. In rat peripheral blood the double positive cells significantly increased in models of Px at the 4th week, and then backed to normal level after MLT supplementation. The results showed no significant changes in all groups at the 8th weeek;At the 4th weeks, Foxp3 expression increased in the thymus of Px rats compared to nomal/sham group, which returned to normal level after MLT supplementation. On the other hand, Foxp3 expression showed no significant difference in all groups at the 8th weeek.Conclusion Px made no difference between the development of CD4~+CD25~+Treg cells in rat thymus, but resulted in significant increase of thymic output and Foxp3 expression. All the effects disappeared at the 8th week. MLT supplementation could reverse the abnormity.

OBJECTIVETo investigate the early changes and clinical significance of plasma endothelin (ET), nitric oxide (NO) and arginine-vasopressin (AVP) in patients with acute moderate or severe cerebral injury.

METHODSThe early (at 24 hours after injury) plasma concentrations of ET, NO and AVP were measured with radioimmunoassay and Green technique in 48 cases of acute moderate (GCS8 in 21 cases) cerebral injury (Group A), in 42 cases of non-cerebral injury (Group B) and in 38 normal individuals (Group C), respectively.

RESULTSThe early plasma concentrations of ET (109.73 ng/L+/-12.61 ng/L), NO (92.82 micromol/L+/-18.21 micromol/L ) and AVP (49.78 ng/L+/-14.29 ng/L) in Group A were higher than those in Group B (67.90 ng/L+/-11.33 ng/L, 52.66 micromol/L+/-12.82 micromol/L and 29.93 ng/L+/-12.11 ng/L, respectively, P<0.01) and Group C (50.65 ng/L+/-17.12 ng/L, 36.12 micromol/L+/-2.16 micromol/L and 5.18 ng/L+/-4.18 ng/L, respectively, P<0.001 ). The amounts of ET, NO and AVP in patients with severe cerebral injury were 116.18 ng/L+/-18.12 ng/L, 108.19 micromol/L+/-13.28 micromol/L and 58.13 ng/L+/-16.78 ng/L, respectively, which were significantly higher than that of the patients with moderate cerebral injury (92.33 ng/L+/-16.32 ng/L, 76.38 micromol/L+/-12.71 micromol/L and 36.18 ng/L+/-12.13 ng/L respectively, P<0.01 ). The early levels of ET, NO and AVP in Group A were negatively related to the GCS scales. The amounts of ET, NO and AVP were 126.23 ng/L+/-15.23 ng/L, 118.18 micromol/L+/-10.12 micromol/L and 63.49 ng/L+/-14.36 ng/L respectively in patients with subdural hematoma, which were significantly higher than those in patients with epidural hematoma (81.13 ng/L+/-12.37 ng/L, 68.02 micromol/L+/-13.18 micromol/L and 45.63 ng/L+/-12.41 ng/L respectively, P<0.01). The plasma concentrations of ET, NO and AVP in stable duration (at 336 hours after injury) in Group A and Group B were similar to those in Group C.

CONCLUSIONSET, NO and AVP were related to the pathophysiological process that occurs in the early stage of acute cerebral injury and the values of ET, NO and AVP correlate positively with the clinical manifestations. The changes of plasma ET, NO and AVP can be regarded as important indices to assess the severity of acute cerebral injury.

Adolescent ; Adult ; Arginine Vasopressin ; blood ; Brain Injuries ; blood ; physiopathology ; Endothelins ; blood ; Female ; Glasgow Coma Scale ; Humans ; Male ; Middle Aged ; Multiple Trauma ; blood ; Nitric Oxide ; blood

Objective　To investigate the early changes and clinical significance of plasma endothelin(ET) and nitric oxide(NO) in patients with acute brain injury.　Methods　The plasma ET and NO concentration at 1 day after injury were checked with radioimmunoassay and Green method in 48 cases of acute brain injury, in 42 cases of non-brain injury and in 38 healthy volunteers. 　Results　 The early plasma concentrations of ET and NO were (109.73±12.61) ng/Lf and (92.82±18.21) μmol/L, respectively, in 48 cases of acute cerebral injury, which were higher than those in the non-brain injury group［(67.90±11.33) ng/L and (52.66±12.82) μmol/L, respectively, P<0.01］ and in the normal control group ［(50.65±17.12)ng/L and (36.12±12.16) μmol/L, respectively, P<0.001］. The amounts of ET and NO in the patients with severe brain injury were (116.18±18.12) ng/L and (108.19±13.28) μmol/L, respectively, which were significantly higher than that of the moderate brain injury group［(92.33±16.32) ng/L and (76.38±12.71) μmol/L, respectively, P<0.01］.The early levels of ET and NO were negatively related to the GCS scores of the patients with acute brain injury. The amounts of ET and NO were (126.23±15.23)ng/l and (118.18±10.12) μmol/L, respectively, in the patients with subdural hematomas, which were significantly higher than those in the patients with epidural hematomas ［(81.13±12.37) ng/L and(68.02±13.18) μmol/L, respectively, P<0.01］. The stable plasma concentrations of ET and NO at 14 days after injury in the patients with acute brain injury and with non-brain injury were similar to that in the normal controls. 　Conclusions　 ET and NO are related to the pathophysiological process in the early period of acute brain injury, and the values of ET and NO correlate positively with the clinical manifestations. The changes of plasma ET and NO can be regarded as important indexes to diagnose the severity of acute brain injury.

Objective:　To study the changes and clinical significance of arginine vasopressin (AVP) and angiotensin II (AT-II) in patients with acute moderate and severe cerebral injury. 　　Methods:　The early plasma concentration was checked by radioimmunoassay in 47 cases of acute moderate and severe cerebral injury, 30 cases of non-cerebral injury and 30 healthy volunteers. 　　Results:　The early plasma concentrations of AVP (50.23 ng/L±15.31 ng/L) and AT-II (248.18 ng/L±82.47 ng/L) in cerebral injury group were higher than those in non-cerebral injury group (AVP for 30.91 ng/L±11.48 ng/L and AT-II for 120.67 ng/L±42.49 ng/L, P<0.01). The early plasma concentrations of AVP and AT-II in cerebral injury group were also obviously higher than those of the volunteers (AVP for 5.16 ng/L±4.23 ng/L and AT-II for 43.11 ng/L±16.39 ng/L, P<0.001). At the same time, the early plasma level of AVP (58.90 ng/L±18.12 ng/L) and AT-II (292.13 ng/L±101.17 ng/L) was higher in severe cerebral injured patients than moderate cerebral injured ones (AVP for 36.68 ng/L±12.16 ng/L and AT-II for 201.42 ng/L±66.10 ng/L, P<0.01). The early level of AVP and AT-II was negatively related to the GCS scales in acute cerebral injury. The early plasma concentrations of AVP (45.98 ng/L±13.48 ng/L) and AT-II (263.28 ng/L±80.23 ng/L) were lower in epidural hematoma group than those of subdural hematoma and cerebral injury group (AVP for 64.12 ng/L±15.56 ng/L and AT-II for 319.82 ng/L±108.11 ng/L, P<0.01). 　　Conclusions:　 AVP and AT-II may play an important role in pathophysiologic process in the secondary cerebral injury. The more severe the cerebral injury is, the higher the early level of AVP and AT-II will be. The early plasma level of AVP and AT-II may be one of the severity indexes of cerebral injury.