Objective To screen and determine the effective silencing targets of β2-microglobulin(B2m)gene at the cellular level in marmoset.Methods By homology comparison of the b2m gene in human and the B2m gene in marmoset, choose homology small hairpin RNA(shRNA)sequences targeting marmoset B2m gene were designed, We choose homology small hairpin RNA(shRNA)sequences targeting designed B2m gene to make homology analysis, and insert into lentivirus-based gene silencing constructs FUGW-TDT.The vectors were transfected into HEK293T cells induced by polyethylenimine(PEI).The suppression of B2m mRNA was detected by real-time PCR.Results Two gene-silencing sequences were screened that lied in 290~310 bp and 665~685 bp of the marmoset B2m mRNA, and have statistical significance in the silencing rate:(46.54±7.91)% (P < 0.05) and(83.22±4.37)%(P < 0.0001).Conclusions Two effective silencing target sequences are screened at cellular level, which can be further used in studies on gene silencing in marmoset.

Objective:To observe the changes in percentages and subsets of invariant nature kiler T (iNKT) cells in adipose and related tissues at different stages of obesity, and analyze the role of iNKT cells during chronic inflammation in adipose tissues in a mouse model of obesity established with high-fat diet.Methods:Changes in mouse body weight, mental state, glucose tolerance and insulin tolerance were recorded. Hematoxylin and eosin (HE) staining was used to observe pathological changes in adipose tissues. Flow cytometry was performed to detect the percentages and subsets of iNKT cells as well as the percentages and subtypes of macrophages. The levels of cytokines in serum samples and the culture supernatants of lymphocytes in adipose tissues were detected with CBA. The expression of related proteins in adipose tissues was detected by Western blot.Results:(1) The volume of adipose cells increased significantly after four weeks of high-fat feeding, but the infiltration of inflammatory cells was not obvious. Significantly increased infiltration of inflammatory cells was observed after 12 weeks of high-fat feeding. (2) High-fat feeding could reduce the percentage of iNKT cells, increase the proportion of iNKT1 subgroup and decrease the proportion of iNKT10 subgroup in adipose tissues. The proportion of iNKT1 subgroup in thymus increased, but that of iNKT2 subgroup decreased. The percentage of macrophages and the proportion of M1 subgroup in adipose tissues increased, while the proportion of M2 subgroup decreased, which were more obvious after 12 weeks of high-fat feeding. (3) High-fat feeding resulted in decreased expression of E4BP4 and arginase-1 (Arg-1) in adipose tissues and increased expression of inducible nitric oxide synthase (iNOS). (4) High-fat feeding significantly increased the pro-inflammatory cytokines and decreased the anti-inflammatory cytokines in mouse serum and culture supernatants of lymphocytes in adipose tissues with more significant changes observed after 12 weeks of high-fat feeding.Conclusions:Increased iNKT1 and decreased iNKT10 in obese adipose tissues might be closely related to the increased M1 polarization and the imbalance of iNKT subsets might be involved in the progression of chronic inflammation in obese adipose tissues.

Erythroderma with complicated etiology is one of the severe skin diseases and has high mortality,of which the incidence was 0.5％-1.5％ in skin diseases.Erythrodermic psoriasis (EP) is the commonest type of erythroderma.In addition,there are drug-induced erythroderma,erythroderma secondary to preexisting dermatoses,malignancy-related erythroderma,and idiopathic erythroderma of unknown etiology.Erythroderma of different etiologies has various clinical manifestations,resulting in relevant curative effects and outcomes.In this article,we retrospectively investigated 205 erythroderma patients about clinical symptoms,auxiliary examination and treatments,and evaluated the efficacy and prognosis.There were 84 cases of EP among 205 patients,10 cases of erythroderma caused by specific drugs,77 cases of erythroderma secondary to preexisting dermatoses (excluding psoriasis),7 cases of erythroderma patients suffering from malignancy and 27 cases with unknown causes.We concluded that the etiology of male patients in different age groups had significant difference.The incidence of EP was the highest among all types.The EP was commonly accompanied with hypoproteinemia,and changed into psoriasis vulgaris after treatment.Drug-induced erythroderma was commonly accompanied with fever,and mostly cured by systematic steroid therapy.For erythroderma secondary to preexisting der matoses,the original dermatoses must be actively treated to achieve a satisfying prognosis.Erythroderma with malignancy or unknown causes had long-term duration,poor response to the treatment,and high potential to relapse.Therefore,clarifying the etiology,providing an appropiate and individual regimen,and regular follow-up are crucial for the successful treatment of erythroderma with unknown causes.

Objective To investigate the effects of mercury on T lymphocytes and serum immune indexes of workers with Methods occupational mercury exposure. A total of 45 workers with occupational mercury exposure were selected as the ， mercury exposure group and 47 workers without occupational mercury exposure were selected as the control group using the judgment sampling method. Cold atomic absorption spectrometry was used to detect the urinary mercury level of the two groups. （ ） +， + +， + + - + Flow cytometry was used to detect the proportion of cluster of differentiation CD 3 CD3CD4 CD3CD8 and CD3CD19 ， - （ - ） - （ - ） cells in peripheral blood and the levels of tumor necrosis factor α TNF α and interleukin 8 IL 8 in serum. The levels of （ ） ， Results immunoglobulin Ig A IgG and IgM in serum were measured by immune nephelometry. The urinary mercury level of （ ： vs ，P ） individuals in the mercury exposed group was higher than that of the control group median 92.7 13.2 μg/g Cr <0.01 . The +， + +， - + proportion of CD3 CD3CD4 CD3CD19 cells in peripheral blood and serum IgG level in the mercury exposed group （ P ）， - - （ P ） decreased all <0.05 and the serum TNF α and IL 8 levels increased all <0.01 compared with the control group. Urinary - + mercury level was negatively correlated with the proportion of CD3CD19 cells in peripheral blood and serum IgG level in the ［ （r） ， ， P ］， study subjects Spearman correlation coefficient S were −0.21 and −0.31 respectively all <0.05 and positively - - （r ， ， P ） ， correlated with serum TNF α and IL 8 levels S were 0.36 and 0.39 respectively all <0.05 . However the urinary mercury （ P ）， +， + +， level was neither correlated with IgA and IgM levels in serum all >0.05 nor with the proportion of CD3 CD3CD4 + + （ P ） Conclusion CD3CD8 cells in peripheral blood all >0.05 . Occupational exposure to mercury can lead to abnormal ， changes in peripheral blood T lymphocyte subsets B lymphocytes and serum immune factors in workers. The mercury load of occupational mercury exposure workers may impact their immune function.

OBJECTIVETo investigate the spectrum of pathogens for community-acquired pneumonia (CAP) in children, and to provide a basis for the diagnosis and treatment of CAP.

METHODSRespiratory secretions and venous blood samples were collected from 1560 children with CAP aged from one month to 9 years within 2 hours after admission, for detection of multiple pathogens. Respiratory virus antigens in nasopharyngeal swab specimens were detected by immunofluorescence. Sputum was used for bacterial culture. Levels of Mycoplasma pneumoniae (MP)-IgM and Chlamydia pneumoniae (CP)-IgM in venous blood were measured by enzyme-linked immunosorbent assay.

RESULTSA total of 579 strains of bacteria were isolated from all respiratory secretions, including 213 (36.8%) Gram-positive strains and 366 (63.2%) Gram-negative strains. The five most common strains were Haemophilus influenzae (7.50%), Streptococcus pneumoniae (6.73%), Staphylococcus aureus (6.35%), Moraxella catarrhalis (5.19%), and Escherichia coli (3.46%), wherein the beta-lactamase-producing strains accounted for 3.3% of all strains. The non-bacterial pathogens mainly included respiratory syncytial virus (12.88%), MP (7.88%), and CP (8.91%). Mixed infection of pathogens was serious, and the mixed infection of respiratory syncytial virus with Haemophilus influenzae infections were the most common. For most pathogens, the infection rate was higher in children aged under one year than in those aged over one year.

CONCLUSIONSHaemophilus influenzae, respiratory syncytial virus, MP and CP are the main pathogens for children with CAP. For most pathogens, the infection rate is higher in children aged under one year than in those aged over one year. Mixed infection rate of pathogens is high.

Child ; Child, Preschool ; Coinfection ; etiology ; microbiology ; Community-Acquired Infections ; etiology ; microbiology ; Female ; Humans ; Infant ; Male ; Pneumonia ; etiology ; microbiology

ObjectiveTo observe andanalyze the clinical efficacy of needle knife treatment for knee meniscus injury from improving knee biomechanical balance and to provide a therapeutic basis and method for clinical practice.MethodOne hundred patients (112 knees) with knee meniscus injury meeting the inclusion criteria were allocated, in order of visits, to observation and control groups, 50 cases each. They were treated with a needle knife and an injection of sodium hyaluronate injectio into knee joint cavity, respectively. Changes in the indices were observed and assessed in the two groups by recording the plantar pressure analysis score and the knee function score before and after treatment and at follow-up. The clinical therapeutic effects were evaluated in the two groups.ResultThemaximum load, time integral difference value and load ratio around the knee joint decreased significantly in both observation and control groups after treatment and at follow-up (P<0.05). The regulating effect on knee joint force balance was better in theneedle knife group than in the control group (P<0.05). The improving effect on knee joint function was significantly better in the observation group than in the control group (P<0.01). The excellence rate was significantly higher in the observation groupthan in the control group after treatment and at follow-up (P<0.05).ConclusionNeedle knife treatment restores the function of the knee joint by loosing focal soft tissues around the knee joint, improving knee joint load imbalance and correcting the mechanical equilibrium around the knee joint in patient with knee meniscus injury. It has a definite curative effect on the disease.

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

Objective:To investigate the mechanism of invariant natural killer T (iNKT)2 cell improving hepatic fat deposition in nonalcoholic fatty liver (NAFL).Methods:NAFL model was established by feeding C57BL/6J mice with high fat diet. The levels of serum total cholesterol, triglyceride, high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in the peripheral blood of mice were analyzed using automatic biochemical analyzer. The pathological changes of liver were observed with HE staining. The cell frequencies of iNKT, iNKT1, and iNKT2 in liver were detected by flow cytometry. Western blotting was used to detect the expression of sterol regulatory element binding protein 1c (SREBP-1c), peroxisome proliferator activated receptor (PPAR)-α, and nuclear factor-κB (NF-κB) in liver tissues.Results:Compared with control group, the body weight of NAFL mice increased, the levels of total cholesterol, HDL-C, LDL-C, ALT, and liver fat deposition increased, the protein expression of SREBP-1c and PPAR-α in liver increased as well as the the protein phosphorylation level of NF-κB. After intraperitoneal injection of α-galactosylceramide (α-GalCer), the levels of total cholesterol, HDL-C, and LDL-C, liver fat deposition decreased, liver SREBP-1c was down-regulated, PPAR-α expression was up-regulated, and the proportion of liver iNKT2 subgroup increased in NAFL mice.Conclusion:iNKT2 cells improve NAFL liver fat deposition, which is related to the down-regulation of SREBP-1c and up-regulation of PPAR-α.

Traditional Chinese Medicine massage is a kind of physiotherapy which affects on specific parts of the body surface by means of training to regulate the function of the body to achieve the therapeutic effect. In this work,under positive detection model, the chemical fingerprint of exhaled breath from volunteers before and after receiving Traditional Chinese Medicine massage within m/z 50-1000 were detected by extractive electrospray ionization-mass spectrometry (EESI-MS). And through high resolution mass spectrometry analysis, the metabolites such as epinephrine (m/z 184. 0889), 3-(3-hydroxyphenyl) propionic acid (m/z 167.0615) and L-tryptophan (m/z 205. 0933) were successfully identified. Besides, chemical fingerprints of volunteers before and after receiving Traditional Chinese Medicine massage under different health condition were clearly differentiated via partial least squares discrimination analysis (PLS-DA). The results showed that Traditional Chinese Medicine massage could significantly change the metabolic process of volunteers. Moreover, it further indicated that the established method could provide a real time fashion to follow metabolic changes caused by Traditional Chinese Medicine massage.

Objective: To evaluate the associative role of depression and apolipoprotein E epsilon 4 allele (APOEε4) in subjective cognitive decline (SCD) and its progression to objective cognitive decline. Methods: After literature search in electronic databases, studies were selected by following precise eligibility criteria. Meta-analyses were performed to examine the role of APOEε4 and depression in SCD or its progression to mild cognitive impairment (MCI) or dementia. Results: APOEε4 positivity was not different between SCD and normal individuals but was significantly higher in individuals with SCD plus than in normal individuals [odds ratio: 2.39 (95% CI: 1.87, 3.05); p<0.00001] and in SCD converters than in non-converters [odds ratio: 5.19 (95% CI: 2.36, 11.42); p<0.00001]. Depression was significantly higher in individuals with SCD [standardized mean difference: 0.63 (0.45, 0.82); p<0.00001] and SCD plus [standardized mean difference: 0.83 (0.43, 1.22); p<0.0001] than in normal individuals. However, depression was not different between SCD and MCI or between SCD converters and non-converters. Age of SCD converters was higher than non-converters [mean difference: 2.95 years (0.58, 5.31)]. Conclusion Whereas APOEε4 positivity was higher in SCD plus and SCD converters, depression was higher in SCD and SCD plus but was not different between SCD and MCI.