OBJECTIVE: To explore the neuroprotective effect of electroacupuncture therapy on damaged cholinergic neurons in hippocampus in aged rats with Alzheimer's disease (AD). METHODS: Thirty six aged male rats were randomly divided into normal control group, sham-operated group, untreated group and electroacupuncture group. Animal model of AD was established with fimbria-fornix transection. The rats in the electroacupuncture group received electroacupuncture on Baihui (DU 20), Yongquan (KI 1), Taixi (KI 3) and Xuehai (SP10). The activity of choline acetyltransferase (ChAT) in septal area of brain was detected by radioimmunoassay, and the protein expressions of nerve growth factor (NGF) and c-fos in CA3 region of hippocampus were detected by immunohistochemical assay. RESULTS: The ChAT activity and the expression levels of NGF and c-fos proteins in the electroacupuncture group were significantly higher than those in the untreated group. CONCLUSION: Electroacupuncture therapy can protect cholinergic neurons in hippocampus in aged rats with AD by means of promoting synthesis of c-fos protein and increasing the expression level of NGF.

Recombinant Fl-V (rFl-V) fusion protein is the main ingredient of the current candidate vaccine against Yersinia pestis infection, which has been under investigation in clinical trial in USA. We investigated the soluble expression conditions of rF1-V in Escherichia coli BL21 (DE3) that we constructed before. After scale-up and optimization of fermentation processes, we got the optimized fermentation process parameters: the culture was induced at the middle exponential phase with 50 µmol/L of IPTG at 25 °C for 5 h. Soluble rFl-V protein was isolated to 99% purity by ammonium sulfate precipitation, ion exchange chromatography, hydrophobic chromatography and gel filter chromatography. The protein recovery was above 20%. Protein identity and primary structure were verified by mass spectrometry and Edman sequencing. Results of purity, quality and western blotting analysis indicated that the target protein is a consistent and properly folded product. Furthermore, the immunogenicity of various antigens formulated with aluminum hydroxide adjuvant was evaluated in mice. Serum antibody titers of 4 groups including 20 µg rFl, rV and rFl-V and 10 µg rFl+10 µg rV, were assayed by ELISA after 2 doses. The antibody titers of anti-Fl with 20 µg rFl-V were obviously higher than titers with other groups; meanwhile there were no significant difference of anti-V antibody titers among them. These findings confirm that rFl-V would be the active pharmaceutical ingredient of the plague subunit vaccine.
Adjuvants, Immunologic ; Animals ; Antibodies, Bacterial ; blood ; Antibody Formation ; Antigens, Bacterial ; immunology ; Blotting, Western ; Chromatography, Ion Exchange ; Enzyme-Linked Immunosorbent Assay ; Mice ; Plague ; prevention & control ; Plague Vaccine ; immunology ; Recombinant Fusion Proteins ; immunology ; Vaccines, Subunit ; immunology ; Yersinia pestis

Reactive oxygen species (ROS) is defined as the electron reduction product of oxygen with high reactivity which can maintain normal physiological functions and redox homeostasis. The tumor microenvironment is in a state of oxidative stress. ROS can affect multiple processes of tumor immune response by modulating the phenotype and functions of tumor cells and immune cells. With the rapid development of immunology, ROS-based tumor immunomodulation has been widely concerned and studied. In this review, the mechanism of ROS participating in tumor immune response is elaborated. Meanwhile, the research process and application of ROS in tumor immunomodulation in recent years are reviewed and analyzed.

Choosing the right solvent and timely use is the basis of rational drug use and the most direct and efficient way to improve the safety of traditional Chinese medicine injections. This study aimed to evaluate the influence of solvent and drug preparation time on Shuanghuanglian injection inducing pseudo-allergic reactions with mouse mode. The two tests were carried out: (1) Comparative experiment between different solvent: Shuanghuanglian injection preparation to the appropriate concentration with 0.9% sodium chloride injection and 5% dextrose injection, mixed with Evans blue, at one time intravenous injected into mice, 30 minutes later, the mouse ears vascular permeability were observed and compared. (2) Comparative experiment among different preparation time: placed 10 min, 2.5 h, 6 h and 24 h after Shuanghuanglian injection were prepared and then to detect the pseudo-allergic reactions in mice using the same methods as in (1). The results showed that there was no significant difference in the pseudo-allergic reactions in mice which induced by the same dose of Shuanghuanglian injection, respectively with 0.9% sodium chloride injection and 5% dextrose injection preparation, and with the extension of preparation time, the degree of pseudo-allergic reactions of Shuanghuanglian injection was gradually severe.
Animals ; Drug Compounding ; Drug Hypersensitivity ; etiology ; Drugs, Chinese Herbal ; adverse effects ; Injections ; Male ; Mice ; Mice, Inbred ICR ; Solvents ; Time Factors

To evaluated the pseudo-allergic reactions of cordate houttuynia, pulse-activating and Qingkailing injection in mice, the ICR mouse were divided into different test groups, then were intravenously injected with three traditional Chinese medicine injections, positive control compound 48/80 and physiological saline as normal control. All test substances were mixed with 0.4% Evans blue. The reaction and vascular permeability of the ears were observed and measured 30 min after injection. At the same time, the mechanisms of the traditional Chinese injections' pseudo-allergic reactions was studyed. ICR mice were injected with the test substances as above without EB, blood in a part of mice were collected after 5 min, and the level of histamine in the plasma were measured. Blood in the other part of mice were collected after 30 min, and the level of VEGF, TNF-α and IL-10 in the serum were measured. The reasults showed that except the cordate houttuynia injection, pulse-activating injection in 1. 5 times clinical concentration and Qingkailing injection in 3.3 times clinical concentration caused mild pseudo-allergic reactions mainly for vascular permeability, no pseudo-allergic reactions occurred when the concentration of the two injections was below the concentration metioned above; 5 minutes after intravenous injection of the three TCM injections into ICR mice with the highest dose, the levels of histamine in plasma of pulse-activating injection and Qingkailing injection groups were increased significantly, 30 minutes later, the levels of VEGF, TNF-α and IL-10 in the serum of all groups were no significant change compared to normal group. The mouse of pulse-activating and Qingkailing injection groups showed inflammatory changes in ear and lung tissues. It can be conluded that when the dose or concentration increased to a certain extent, pulse-activating and Qingkailing injection could induce pseudo-allergic reactions on ICR mice, the increased histamine realease maybe the main reason for pseudo-allergic reactions of the two traditional Chinese medicine injections. In addition the author preliminary thought that inflammatory mechanisms leading to hyperpermeabilities were the main reason of the traditional Chinese medicine injection's pseudo-allergic reaction.
Animals ; Drug Hypersensitivity ; etiology ; Humans ; Injections ; Interleukin-10 ; blood ; Male ; Medicine, Chinese Traditional ; adverse effects ; Mice ; Mice, Inbred ICR

OBJECTIVETo study the correlation between apparent diffusion coefficient (ADC) and Ki-67, a marker of tumor activity, in the diagnosis of gliomas.

METHODSConventional magnetic resonance imaging (MRI), enhanced scanning, and diffusion-weighted imaging were performed in 76 patients with pathologically confirmed gliomas. The ADC values were measured at tumor parenchyma and the corresponding contralateral normal brain. The relatively ADC (rADC) values of the tumor parenchyma were calculated. The correlation of the ADC values with tumor grades was analyzed. The expression of Ki-67 was detected by immunohistochemical staining. The correlation between ADC value and Ki-67 in the diagnosis of gliomas was analyzed.

RESULTSThe ADC values and rADC values of high-grade gliomas were significantly lower than those of low-grade gliomas. The ADC values of tumor parenchyma were inversely associated with the degree of malignancy of the gliomas (r=-0.898, r=-0.868; P<0.01). The expression of Ki-67 was significantly higher in high-grade gliomas than that in low-grade gliomas. The Ki-67 labeling index in grade 3 and 4 gliomas were (29.48 ± 19.78)% and (31.21 ± 17.50)%, respectively. Both of them were significantly higher than Ki-67 labeling index in low-grade (grade 1 and 2) gliomas [(2.33 ± 2.20)%] (P<0.01). Nevertheless, the Ki-67 labeling index showed no significant difference between grade 3 and 4 gliomas (P>0.05). The expression of Ki-67 was negatively correlated with the ADC values and rADC values in tumor parenchyma (r=-0.627, r=-0.607; P<0.01).

CONCLUSIONThe ADC and rADC values of tumor parenchyma can indirectly reflect the proliferation and malignancy of gliomas and therefore can be useful for the grading of glioma.

Adolescent ; Adult ; Brain Neoplasms ; diagnosis ; metabolism ; pathology ; Child ; Child, Preschool ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Glioma ; diagnosis ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Male ; Middle Aged ; Young Adult

OBJECTIVETo study the effect of Wenyang Decoction (WD) on the differentiation of CD34+ progenitor cells of occupational asthma (OA) model rats.

METHODSFifty healthy male SD rats were randomly divided into five groups, i.e., the model group, the blank control group,the WD group,the Western medicine group,the combined group, 10 in each group. Prednisone suspension (10 mg/kg) was administered to rats in the Western medicine group by gastrogavage. WD (20 g/kg) was administered to rats in the WD group by gastrogavage. Prednisone suspension plus WD was administered to rats in the combined group by gastrogavage. Normal saline was administered to rats in the model group and the blank control group by gastrogavage. The general condition of rats was observed. Expression levels of peripheral blood IL-5 and eotaxin, eosinophils (EOS), CD34+, CC chemokine receptor 3 (CCR3+) in bone marrow suspension were detected by ELISA, Wirght-Giemsa, and flow cytometry, respectively.

RESULTSCompared with the blank control group,expression levels of IL-5 and eotaxin in peripheral blood were significantly higher (P < 0.01), and the count of EOS and CD34+ cells, as well as CD34+ /CCR3+ significantly increased (P < 0.01) in the model group. Compared with the model group, expression levels of IL-5 and eotaxin, the count of EOS, CD34+ cells, CD34+ / CCR3+ were lowered in three treated groups (P < 0.01). Compared with the Western medicine group, the count of EOS and CD34+ / CCR3+ decreased in the combined group (P < 0.01). The count of EOS was significantly lower in the combined group than in the WD group (P < 0.01).

CONCLUSIONWD could reduce levels of in vivo inflammatory factors, and restrain the differentiation and recruitment of EOS,thereby alleviating the differentiation of CD34 progenitor cells to EOS.

Animals ; Antigens, CD34 ; Asthma, Occupational ; drug therapy ; Bone Marrow ; Cell Differentiation ; Chemokine CCL11 ; Drugs, Chinese Herbal ; therapeutic use ; Eosinophils ; Flow Cytometry ; Interleukin-5 ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, CCR3 ; Stem Cells

Objective To investigate the incidences, risk factors, genotypes and epidemiology of community-acquired blood stream infection caused by extended spectrum β-lactamases (ESBLs)-producing Escherichia coli and Klebsiella pneumonia strains and to analyze the sensitivity of those ESBLs producing strains to commonly used antibiotics. Methods Forty-two patients who were diagnosed with community-ac-quired blood stream infection caused by Escherichia coli or Klebsiella pneumonia strains in Sichuan Provincial People′s Hospital were recruited in this study. Disc diffusion method was used for the phenotypic confirmato-ry test of ESBLs. Agar dilution method was performed to measure the antimicrobial susceptibility of the ESBLs-producing strains to 13 clinically commonly used antibiotics. Genotypes of the ESBLs-producing strains were identified by polymerase chain reaction (PCR). Multilocus sequence typing (MLST) was used to analyze the epidemiology of ESBLs-producing strains. Logistic regression analysis was performed to analyze the risk factors for community-acquired blood stream infection. Results The ESBLs-producing Escherichia coli strains accounted for 56. 3% (18 / 32) and the ESBLs-producing Klebsiella pneumoniae strains accounted for 20% (2 / 10). All of the 20 ESBLs-producing strains were sensitive to imipenem, meropenem, ertapen-em, nitrofurantoin and moxalactam. The ESBLs-producing strains sensitive to amikacin, piperacillin-tazobactam and fosfomycin accounted for 95% , 90% and 85% , respectively. Drug resistance rates of the 20 strains to cefotaxime, levofloxacin, ciprofloxacin and cefepime were relatively high accounting for 100% , 80% , 80% and 75% , respectively. Among the 20 ESBLs-producing strains, 7 strains only carried the CTM gene, while the other 13 strains were all positive for two genotypes of ESBLs, mainly identified as TEM+CTM-M-14 and TEM+CTM-15 genotypes. The 18 Escherichia coli strains were classified into 10 ST types, most of which were ST131 type, followed by ST10 and ST38 types. This study indicated that malignant tumor might be a possible risk factor. Conclusion The prevalence of community-acquired blood stream infection caused by ESBLs-producing Escherichia coli strains was becoming increasingly serious. Malignant tumor might be the risk factor associated with the producing of ESBLs in Escherichia coli and Klebsiella pneumonia strains. TEM+CTX-M-14 was the predominant genotype of ESBLs-producing strains and the prevalent clone was ST131 type. Carbapenems and enzyme inhibitor compounds were ideal drugs for the treatment of commu-nity-acquired blood stream infection caused by ESBLs-producing Escherichia coli and Klebsiella pneumonia strains. This study was limited by the small sample size. Therefore, it is necessary to conduct further resear-ches based on a large number of samples.

Objective To study the expression of co-stimulatory molecules, GL50-ICOS, in thyroid tissue of patients with Graves' disease (GD) and to explore their relationship with the immune pathogenesis of GD.Methods RT-PCR, Western blot, immunohistochemistry were applied to detect the expression of GL50-ICOS in thyroid of GD. Thyrocytes were cultured in the absence or presence of pro-inflammatory cytokines. The expression of GL50 on thyroid follicular cells (TFC) was further measured by flow cytometry. Results (1) In GD patients,the percentage of CD4+ CD28- T cells was significantly increased as compared with the control healthy individuals. The expression of co-stimulatory molecule ICOS was up-regulated. (2) The mRNA level of ICOS was significantly increased in GD patients than that in nontoxic goiter(NTG) patients(P＜0.01). (3)Compared with NTG control group, the GL50 protein expression was much higher in thyroid tissues of GD patients (P ＜0.01). (4)The results of immunohistochemistry showed that GL50 expression was observed in all GD thyroid tissues, while no expression of GL50 was detected in NTG thyroid tissues(P＜0. 01). (5) The expression of GL50on primary cultured thyroid follicular cells was significantly increased under the stimulatation of pro-inflammatory cytokines in vitro. Conclusion GL50-ICOS is expressed abnormally in thyroid tissue of patients with GD.

Aging can cause degenerative changes in the function of multiple tissues and organs in the body. Gastrointestinal diseases and intestinal dysfunction are very common in the elderly people. The purpose of this study is to explore the effect of the total extract of Astragalus membranaceus (Fisch.) Bge. on intestinal function and gut microbiota homeostasis in natural aging mice, which will provide clues for further mechanism study. The natural aging mice model is established and animal experiments follow the regulations of the Animal Ethics Committee of Nanjing University of Traditional Chinese Medicine. The overall health of the mice was evaluated by the "frailty index" scoring method. The intestinal absorption and transport function were measured by detecting intestinal glucose absorption capacity, transport time, lipase and amylase activities of aging mice. Intestinal inflammation was assessed by detecting inflammatory cytokines by enzyme-linked immunosorbent assay (ELISA). The pathological changes in the intestines of aging mice were tested by hematoxylin-eosin (H&E) staining and alizarin blue (AB) staining. The qRT-PCR method was used to explore the gene transcription level related with the proliferation and differentiation of intestinal stem cells. Microbiota analysis based on 16S rDNA were used to evaluate the composition of gut microbiota. The results showed that Astragalus had a tendency to reduce the "frailty index" of aging mice, but did not show a significant difference. In some indicators of aging phenotype, Astragalus has the most significant effect on hair loss and physical fitness. In terms of intestinal function, Astragalus could increase intestinal glucose absorption capacity, shorten intestinal transportation time and promote lipase secretion in aging mice. The levels of inflammatory cytokines such as tumor necrosis factor-α (TNF-‍α) in the aging intestinal tissue were reduced after Astragalus administration. Astragalus also ameliorated the pathological degeneration of the intestinal tissue of aging mice by increasing the length of small intestinal villi, the thickness of colonic mucosa and goblet cell number. In addition, Astragalus elevated the expression of genes associated with the proliferation and differentiation in jejunum and modulated gut microbiota, especially restoring the abundance of Lachnospiraceae. Taken together, the above research results demonstrate the total extract of Astragalus as a key factor improving the intestinal function and gut microbiota homeostasis of aging mice.