Objective To explore the effect of glutamine(Gln) on expression of nuclear factor kappa B(NF-?B) and heat shock protein 70(HSP70) in brain of young rats induced by lipopolysaccharide(LPS).Methods Ten days old Wistar rats were randomly divided into 3 groups by injection intraperitoneally different agonts,LPS group,normal saline control group(NS group) and Gln group(Gln 1.346 g/kg,1 hour before LPS).NF-?B and HSP70 distribution and expression in brain were deteted by immunohistochemistry.The levels of HSP70 in rats brain induced by LPS were detected by Western blot.SPSS 12.0 software was used.Results The nuclei of neuron in cerebral cortex in LPS group obviously cleared at 6 hours.The positive stain of nuclei in Gln group at 2 hours could not be seen.The stain of nuclei in cerebral cortex was weakened in LPS group at 6 hours by immunohistochemistry.HSP70 protein expression decreased with the measurement of Western blot,especially at 24 hours.HSP70 expression in LPS group was similar as that in NS group.The stain of nuclei in neuron in Gln group at 2 hours increased.It also showed the amount of protein expression increased in Western blot in group Gln at 2,6,12,24 hours(Pa

To study the identification of Gentianaceae Mongolian medicine Digeda with spectroscopy techniques, near infrared spectroscopy and differential scanning calorimetry techniques were applied to study on the identification of 4 kinds of Gentianaceae Mongolian medicine Digeda, and characteristic spectrums obtained were systematically analyzed. In NIR study, the four species of Digeda exist some differences in 4 250-4 400 cm(-1) and 5 650-5 800 cm(-1) of one-dimensional spectra, and show significant differences in 4 100- 4 400 cm(-1), 4 401-4 900 cm(-1) and 5 400-5 800 cm(-1) of the second derivative spectra. DSC curves of them present distinct topological pattern, characteristic peak and peak temperature. Using near infrared spectroscopy and differential scanning calorimetry analysis can realize efficient and accurate identification of four kinds of Mongolian medicine Digeda, and provide scientific basis for the efficient and accurate identification of other Gentianaceae Mongolian medicine Digeda.
Calorimetry, Differential Scanning ; methods ; China ; Gentianaceae ; chemistry ; classification ; Medicine, Mongolian Traditional ; Spectroscopy, Near-Infrared ; methods

ObjectiveTo explore the effect of blocking the JAK/STAT signal pathway on the activity of Caspase-3 in the synovial tissue of rheumatoid arthritis rats.MethodsFifty rat models of collageninduced arthritis,which had arthritis index more than 2 were divided into the model group,the low dosage of AG490 group,the medium dosage of AG490 group and high dosage of AG490 group.Inaddition,6 rats were treated as normal control group.Normal saline,AG490 1,5,10 mg·kg-1 ·d-1 were given by intraperitoneal injection.Then the volume claws and pathologic scores of the rat models were recorded and the activity of Caspase-3 in the synovial tissue were compared.The results were analyzed by one-way ANOVA and LSD-t or Tamhane's T2 test.ResultsThe arthritis of the CIA models progressed fast,the volume of the claws and the pathological score of them were significantly higher than those of the control group.At the same time,no Caspase-3 positive express could be detected in the control group,whilethe model group had slightly increased expression.After different dosages of AG490 were applied,the swollen of joints was significantly improved compared with the model group.The histopathological score of the medium AG490 dosage of group and high dosage group(2.7±0.8,1.8+0.9) were remarkably decreased than those of the model group(4.3+1.2),the differences were statistically significant (P＜0.01).In addition,the Caspase-3 expression in the low,medium and high AG490 dosage group ( 1.90±0.15,3.13±0.33,3.56+0.34) was significantly higher than that of the model group(1.48±0.18)(P＜0.05 or P＜0.01 ).ConclusionBlocking JAK/STAT signal pathway can increase the activity of Caspase-3,reduce the excessive proliferation of synovial tissue,and improve arthritis symptoms.

Sleep disorders, which have high incidence and complicated mechanisms, are a prevalent public health problem that cannot be ignored. It is confirmed that the drug therapy and cognitive behavioral therapy are the main ways to treat sleep disorders effectively. However, these therapies do have limitations. For example, the tolerance and dependency of drug therapy means that it cannot be taken for a long period of time, as well as the cognitive behavioral therapy is greatly influenced by location and economy factors. Thus, computerized cognitive behavioral therapy (CCBT) comes into being, with its convenience and economic advantages. Currently, the CCBT is systematic in some countries, which is still in the beginning stage in China. The application and efficacy of CCBT and the existing problems were reviewed in this paper.

Objective: To observe the clinical efficacy of DZWJY-1 type electronic moxibustion apparatus and traditional moxibustion in treating knee osteoarthritis (KOA). Methods: A total of 76 eligible patients were randomized into an electronic moxibustion apparatus group and a traditional moxibustion group, with 38 cases in each group. The electronic moxibustion apparatus group was intervened by DZWJY-1 type electronic moxibustion apparatus, and the traditional moxibustion group received moxa stick moxibustion for treatment. Neixiyan (EX-LE 4), Dubi (ST 35), Xuehai (SP 10) and Liangqiu (ST 34) were selected for both groups and the treatment was conducted 3 times a week for a total of 12 times. The visual analog scale (VAS) and the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) scores were observed before treatment and after 6 and 12 sessions of treatment, respectively. Results: There were 4 dropout cases in the traditional moxibustion group. Therefore, this trial had 72 valid cases, including 38 cases in the electronic moxibustion apparatus group and 34 cases in the traditional moxibustion group, the differences in the baseline data between the two groups were statistically insignificant (P>0.05). After 6 and 12 sessions of treatment, the VAS scores decreased significantly with the increase of treatment sessions in both groups (all P<0.01), and the betweengroup differences were statistically insignificant at the same time points (both P>0.05). The pain intensity was evaluated using the weighted value of VAS score. The markedly effective rate was 47.4％ and the total effective rate was 89.5％ in the electronic moxibustion apparatus group, versus 50.0％ and 94.1％ in the traditional moxibustion group, and the betweengroup differences were statistically insignificant (both P>0.05). After 6 and 12 sessions of treatment, the total score and the component scores including pain, stiffness and difficulty moving in the WOMAC decreased significantly with the increase of treatment sessions in both groups (all P<0.01), and the between-group differences were statistically insignificant (all P>0.05). Conclusion: Electronic moxibustion apparatus and traditional moxibustion both are effective in reducing joint pain and improving joint function in KOA patients, and they are equivalent comparing the clinical efficacy.

Objective To knockout Rag2 and IL2rg genes and construct severe combined immunodeficiency mice based on CRISPR/Cas9 technology. Method Design and synthesis of 25 bp sgRNA were made according to the Rag2 and IL2rg sequences in Genbank. After annealing, sgRNA was cloned into pX330 vector. Recombination plasmid Rag2?sgRNA, IL2rg?sgRN and Cas9 were then transcribed into RNA, these RNA were microinjected into zygotes and the zygotes were transplanted into recipient ICR mice. F0 founders were born and mutated F0 founders mated with wild type mice to obtain F1 generation heterozygous mice. Mutated F1 mice were crossed and got F2 generation homozygous mice. Genotype and phenotype of the knockout mice were identified by sequencing, flow cytometry and xenograft model. Results Rag2?sgRNA and IL2rg?sgRNA recombination plasmids were constructed and transcribed into RNA. After microinjection and mat? ing, F0 founders were born and F2 homozygous mice were obtained. The results of sequencing showed that there were two types of genotype in IL2rg gene, 10 bp or 11 bp deletion;however, there was only one genotype in Rag2 gene, which was 8 bp deletion. Compared with wild?type BALB/c mice, the number of CD3 +, B220 + and NKp46 + cells in peripheral blood of the knockout mice was reduced significantly. After inoculation of human breast cancer cell line SKBR?2HL cells, tumor size in the xenograft mouse model was increased gradually along with time extension. Conclusion CRISPR/Cas9 is an efficient way to mutate Rag2 and IL2rg gene in mice in vivo, leading to aberrant T cells, B cells and NK cells.

The purpose of this study is to prepare galactosyl modified lipid bilayer-coated mesoporous silica nanoparticles (GPEM) to enhance the antitumor efficacy against hepatocellular carcinoma cells. The irinotecan (CPT-11) loaded mesoporous silica nanoparticles (MSNs) was coated with the Gal-P123 modified functional lipid bilayer by thin-film dispersion method. Nanoparticles were characterized with particle size, zeta potential, morphology and drug release in vitro. Afterwards, the cell uptake, intracellular concentration of CPT-11, cell apoptosis rate and cytotoxicity were evaluated on human hepatocellular carcinoma cell line Huh-7. The results showed that MSNs were coated with intact lipid bilayers and the nanoparticles had clear core-shell structure. GPEM is stable with the mean particle size of (78.01 +/- 2.04) nm. The low leakage rate in normal physiological conditions in vitro is contributed to the protection of stable lipid bilayer, and the fast drug release in acid environment due to the destruction of the lipid bilayer. On the cell level, the vector could improve the intracellular CPT-11 concentration by 4 times because of the functional lipid bilayer. The high CPT-11 concentration led to the increasement of apoptosis rate by 48.6%, and the reduction of half maximal inhibitory concentration (IC50) values of CPT-11 by 2 times, indicating stronger cell cytotoxicity.
Antineoplastic Agents ; chemistry ; pharmacokinetics ; Apoptosis ; Camptothecin ; analogs & derivatives ; chemistry ; pharmacokinetics ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Drug Carriers ; chemistry ; Drug Delivery Systems ; methods ; Humans ; Lipid Bilayers ; chemistry ; Liver Neoplasms ; drug therapy ; pathology ; Nanoparticles ; chemistry ; Particle Size ; Silicon Dioxide ; chemistry

A novel white-rot fungus AH28-2,which was isolated from 224 fungi samples,ability to produce effectively laccase by induction.Several factors influencing laccase production were investigated.The optimum conditions were as follows:the 300mL Erlenmeyer flask containing 150mL of liquid medium was inoculated with 7.5mL of mycelial fragments and the medium was supplied with lignin at a concentration of 0.1%(initial pH8.5).The cultures was incubated at 28℃ on rotary shaker(150r/min) for 4～5 days.The maximum enzyme activity was 20184IU/L.

Objective To explore the safety of the mobilization of peripheral blood stem cells(PBSC)by granulocyte colony-stimulating factor(G-CSF)in elderly donors.Methods 28 peripheral arteriosclerotic occlusive disease(PAOD)elderly patients(aged≥60 years),and 29 healthy sibling young/adult donors(aged＜60 years)for peripheral allogenic stem cell transplantation were included.Blood samples were collected immediately before starting G-CSF and prior to PBSC collection to analyze the following parameters:the WBC counts,fibrinogen(FIB),D-dimer (D-D),thrombin antithrombin complex(TAT),antithrombin(AT)and yon Willebrand factor antigen(vWF:Ag). Results It had a very significant increase in D-D and vWF:Ag and a very significant decrease of AT(P＜0.01),af- ter mobilization by G-CSF,and a increase in FIB and TAT were also observed(P＜0.05,P＜0.01)in elderly group.In the young/aduh group,the increase in FIB was significant(P＜0.05).The elevating extent of D-D and TAT after G-CSF administration was significantly higher in elderly group than that in young/adult group(P＜0.05).Compared to young/adult group,there was a significant increase in thrombotic events and cerebrovascular ac- cident(P＜0.05).Conclusion In PBCS donorsreceiving G-CSF it reveals activation of both coagulation and en- dothelial cells and inhibition of anticoagulant system that can favor the developing of thrombotic events,which is more remarkable in elderly donors.Therefore a careful monitoring for coagulation system should be considered in those elderly cases.

OBJECTIVETo explore the silencing effects of RNA interference on the expression of 3-phosphoinositide-dependent protein kinase 1 (PDK1) gene, and the effects on malignant phenotypes of esophageal carcinoma EC9706 cells.

METHODSPDK1 siRNAs was transfected into the EC9706 cells. The expression of PDK1 mRNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR). At the same time, expressions of PDK1, Akt and phosphorylated Akt proteins were detected by Western blot. Methyl thiazolyl tetrazolium assay (MTT) was used to examine the cell proliferation after transfection. Flow cytometry was used to determine the percentage of apoptosis cells, and Transwell chambers were used to detect the invasion ability of the cells. Tumor formation in nude mice was used to assess the tumorigenic characteristics in vivo.

RESULTSCompared with the non-transfected group, PDK1 siRNA effectively inhibited the expression of PDK1 mRNA in EC9706 cells, with an inhibition rate of (28.5 ± 4.2)% at 24 h, (51.1 ± 5.7)% at 48 h and (60.6 ± 4.1)% at 72 h after transfection. The expressions of PDK1 and phosphorylated Akt protein were also knocked down by PDK1 siRNA (P < 0.05). PDK1 siRNA significantly inhibited the cell proliferation and invasion, promoted the cell apoptosis, and inhibited the EC9706 cells proliferation in vivo and the expression of PDK1 protein in the transplanted tumors (P < 0.05).

CONCLUSIONPDK1 may play an important role in esophageal cancer cell proliferation, invasion and apoptosis, and may serve as an effective target for cancer gene therapy.

3-Phosphoinositide-Dependent Protein Kinases ; Animals ; Apoptosis ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Esophageal Neoplasms ; genetics ; metabolism ; pathology ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Phosphorylation ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Tumor Burden