OBJECTIVETo observe the effect of Shen warming Pi strengthening method on expressions of serum T cell subsets (C045+%, C03+%, and C04 +/COB+) in diarrhea-predominant irritable bowel syndrome (IBS-0) rats. Methods An IBS-0 rat model was established referring to AL-Chaer's modeling method combined with tail clamp and intragastric administration of sanna leaf. After modeling 30 SO rats were randomly divided into 6 groups according to random digit table, i.e., the model group, the high, middle, low dose Wenshen Jianpi Recipe (WJR) groups, and the Sishen Pill control group, 6 in each group. A normal control group consisting of 6 SO rats were also set up. Rats in high, middle, low dose WJ R groups were administered by gastrogavage with boil-free WJ R at the daily dose of 3. 100, 1. 550, 0. 775 g/kg, respectively. Rats in the Sis hen Pill control group were administered by gastrogavage with boil-free Sis hen Pill at the daily dose of 0. 736 g/kg. Equal volume of normal saline was given by gastrogavage to rats in the model group and the normal control group. All medication lasted for 2 successive weeks. Rats' general state, expressions of T cell subsets (CD45+%, CD3+%, and CD4+ /CDB+) changes were observed.

RESULTSCompared with the normal control group, expressions of CD45+% and CD3+% increased, but CD4+ /CDB+ decreased with statistical difference (P < 0. 05). Compared with the model group, expressions of CD45+% and CD3+% decreased, but CD4+ ICDB+ increased with statistical difference in high, middle, low dose WJR groups, and the Sis hen Pill control group (P <0. 05). Compared with the Sis hen Pill control group, there was statistical difference in all indices except CD45+ value in the low dose SWPSM group (P <0. 05). Compared with the low dose WJ R group, the expression of CD3+% decreased in high and middle dose WJR groups, and the Sis hen Pill control group; CD4+ /CD8+ increased in the Sishen Pill control group and the high dose SWPSM group (all P < 0. 05).

CONCLUSIONSWJR showed better treatment effect. The mechanism of Shen warming Pi strengthening method might be achieved by regulating expressions of CD45+% and CD3+%, and CD4+ /CD8+ ratios.

Animals ; Drugs, Chinese Herbal ; Female ; Irritable Bowel Syndrome ; therapy ; Leukocyte Common Antigens ; metabolism ; Medicine, Chinese Traditional ; Rats ; T-Lymphocyte Subsets ; metabolism

Objective To observe the effects of low dose irradiation (LDR) on proliferation,adipogenesis and osteogenic potential of human umbilical cord mesenchymal stem cells (hucMSCs).Methods hucMSCs were isolated from Wharton's jelly tissue of human umbilical cord by modified tissuepiece inoculation,and flow cytometry was used to detect the expression of specific marker in the hucMSCs.The hucMSCs were randomly divided into two groups:irradiation group undergoing irradiation with the doses 50,100,or 200 mGy respectively,and control group without irradiation.MTT method was applied to evaluate the proliferation of the hucMSCs at different time points with various doses irradiation.The third passage hucMSCs were randomly divided into two groups:irradiation group undergoing low dose irradiation of 200 mGy,and control group without irradiation,and then underwent induction by adipocytic and oesteocytic differentiation induction fluids respectively so as to differentiate into adipocytes and osteoblasts.Oil red O staining was used to detect the activity of alkaline phophatase (ALP),and RT-PCR was used to detect the mRNA expression of core binding factor alpha 1 in human osteoblast.Results After 9-12days,fibroblasts began to swim out of the tissue piece with a confluence rate of 80% 2 weeks later.Within 7 days the absorption values of the hucMSCs undergoing different irradiation doses 2,3,4,5,and 6 days later were all significantly higher than those of the control group(F = 159.17,448.81,265.15,183.93,and 181.83 ,all P <0.01),with the proliferation rates of the 100 mGy subgroup being the highest.After being induced liquid,vacuoles were observed in the irradiated group 2 days later.21 days later,the adipogenic rates of irradiated group was significantly higher than that of the control group (t = 28.25,P <0.01).The ALP activity increased in the irradiated group compared with control group (t=16.87,P <0.01) .The expression level of Cbf-α1 mRNA was up-regulated obviously (t = 14.16,P＜0.01).Conclusions LDR promotes the proliferation of hucMSCs,and accelerates the hucMSCs' differentiation into adipocytes and osteoblasts.

ObjectiveTo investigate the cytotoxicity and gene transfection mediated by NMPCS/DNA nanoparticles.MethodsN-methylene phosphonic chitosan (NMPCS) was synthesized using one-step reaction under homogeneous conditions.The NMPCS/DNA nanoparticles were prepared using complex coacervation method.The cytotoxicity of NMPCS alone and its complexes with plasmid DNA were determined by MTT assay on HeLa cells.The gene transfection mediated by NMPCS/DNA nanoparticles were investigated using pGL3control vector as reporter gene.ResultsThe MTT results suggested that the NMPCS and NMPCS/DNA complexes showed significantly lower cytotoxicity than PEI and PEI/DNA complexes,respectively.The gene transfection mediated by NMPCS/DNA nanoparticles were greatly improved compared with unmodified chitosan.ConclusionNMPCS would demonstrate great potential as a novel,safe,efficient non-viral vector for gene delivery.

Objective To explore the gene expression of interleukin-13 receptor (IL-13R)?2 and its relationship to proliferation activity of human gliomas.Methods The gene expression of IL-13R?in 50 hu- man gliomas,2 malignant human glioma cell lines and 6 normal brain tissues were studied by RT-PCR.Ki-67 labeling index (Ki267 LI) of all sample were detecteded by immunohistochemical staining.Results Only one normal brain tissues expressed very low IL-13R?2 mRNA,whereas 35 (70%) of 50 human brain tumors expressed 1L-13R?2 mRNA.The positive rate and expression level of IL-13R?2 mRNA were increased with the ascending of WHO tumor grade.(former:rs=0.87;letter:rs=0.69,P＜0.01).The difference of posi- tive rate and expression level of IL-13R 2?mRNA between the low grade and high grade tumors was statistical- ly significant,the proliferation activity of gliomas evaluated by Ki-67LI (Ki-67 Labeling Index,Ki-67LI) was positively correlated with IL-13R?2 gene expression and the tumor grade.Conclusion In human cerebral gliomas,IL-13R?2 genes may play an role in the malignant progression.The expression level of malignancy in molecular level and selecting the target of gene therapy.

Carcinoma, Squamous Cell ; genetics ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; genetics ; pathology ; virology ; DNA, Viral ; metabolism ; Female ; Gene Amplification ; Humans ; In Situ Hybridization, Fluorescence ; Lymphatic Metastasis ; Neoplasm Grading ; Neoplasm Staging ; Papillomaviridae ; genetics ; Papillomavirus Infections ; metabolism ; RNA ; genetics ; Telomerase ; genetics ; Uterine Cervical Neoplasms ; genetics ; pathology ; virology

Objective To explore the activated brain region of acute epilepsy in cat model induced by pentylenetetrazol(FFZ)with manganese enhanced-functional MR imaging(ME-fMRI),and evaluate the application of ME-fMRI on localization of the activated brain.Methods Forty cats were divided into 4 groups by random number table method as epileptic A and B groups as well as control A and B groups. Cats of epileptic groups were injected with PTZ(55 mg/kg)intramuscularly,and those of control groups were injected with the saline at same dose.The behavior change in the epileptic and control group A was observed and electroencephalogram(EEG)was also undertaken.Cats of epileptic and control group B were performed ME-fMRI,and the percentage of the enhanced signal intensity was then calculated.Results After injection with PTZ(55 mg/kg)intramuscularly,epileptic seizure was all evoked,and then EEG recording showed spike-wave and polyspike-wave complexes.The neocortex of cats of epileptic group B was diffusely phanero-enhanced on ME-fMRI.The percent enhancement of signal intensity in cortex of frontal lobe,parietal lobe and occipital lobe was(34.6?5.7)% and that in cortex of temporal lobe with(22.9? 6.5)%,whereas those of control group B with(14.9?4.5)% and(11.6?3.2)% respectively.And there was significant difference between the above different localization of the brain in the two groups (t=-10.43,-5.46 respectively,P

OBJECTIVEIBS-D rat model was established to assess the effect of Shen warming Pi strengthening method (SWPSM) for intervening diarrhea-predominant irritable bowel syndrome (IBS-D) by observing rats' general state, stool properties, AWR ranking, and histopathological changes.

METHODSTotally 72 rats were randomly divided into 6 groups, i.e. the normal group, the model group, the high, middle, low dose SWPSM groups, and the control group, 12 in each group. The IBS-D rat model was successfully established referring to AL-Chaer ED's modeling method. After modeling high, middle, and low dose SWPS Recipe boil-free granules were given by gastrogavage to rats in corresponding treatment groups. Sishen Pill boil-free granule was given by gastrogavage to those in the control group. Equal volume of normal saline was given by gastrogavage to rats in the model group. The medication lasted for 2 weeks. Rats' general state, stool properties, abdominal withdrawal reflex (AWR) ranking, and histopathological changes were observed.

RESULTSAfter treatment, the general state of all rats got im- provement to various degrees. The improvement in the high and middle dose SWPS Recipe groups were superior to that in the low dose SWPS Recipe group and the control group (P < 0.05). There was no statistical difference in the growth rate between after and before treatment in each group (P > 0.05). Compared with the model group and the low dose SWPS Recipe group, the defecation amount within 4 h was less in the high and middle dose SWPS Recipe groups and the control group (P < 0.05). The Bristol ranking score, average ranking of loose stool, ratio of dry stool and wet stool were lower in the high and middle dose SWPS Recipe groups than in the control group and the low dose SWPS Recipe group (P < 0.05). The AWR ranking score was lower in the high and middle dose SWPS Recipe groups than in the control group when the volume of balloon dilation was 1.5 mL. There was no organic change of histological or morphological observation.

CONCLUSIONSHigh sensitive IBS-D model was proved to be reliable. SWPSM could reduce the quantity of stools, lower Bristol ranking score, average ranking of loose stools as well as ratios of dry stool and wet stool, contributing to reducing the high sensitivity of rats' visceral organs to some extent.

Animals ; Diarrhea ; drug therapy ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Irritable Bowel Syndrome ; drug therapy ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

Objective The immunotoxins generated by conjugating monoclonal antibody (mAb) and a certain toxin play an important and promising role in treating hematopoietic malignancies. However, most of the toxins used for the conjugation are toxic proteins, which are immunogenic in the patients. Norcantharidin (NCTD) is a small molecule toxin without immunogenicity, and thus has become a potential new drug for hematopoietic cancers. In this study, we prepared immunotoxin 2E8-NCTD by using the ZCH-4-2E8 cells produced in the laboratory of our hospital, and then detected its targeting effect against CD+19 lymphoid malignant Nalm-6 cells in vitro.Methods 2E8 mAb was obtained from mouse ascites and purified by gel chromatography. After its purity was checked by SDS-PAGE, immunotoxin 2E8-NCTD was generated by conjugating CD19 mAb with NCTD using activated ester method. The binding activity of the immunoconjugate to CD19 antigens on cell surface, and the expression levels of the CD19 antigens on Nalm-6 and K562 cells were determined by flow cytometry. The inhibitory effects of PBS, purified 2E8 mAb, NCTD, and immunotoxin 2E8-NCTD on the cell growth of either Nalm-6 or K562 cells were then compared.Results The purity of the 2E8 mAb was higher than 99% demonstrated by SDS-PAGE assay. 2E8 mAb was detected on the surface of 99.34% of the Nalm-6 cells, while on only 0.98% of the K562. The newly generated immunotoxin had a positive rate of 99.90% on the Nalm-6 with slightly reduced binding activity. Both 2E8-NCTD and NCTD significantly inhibited the growth of CD+19 Nalm-6 cells (P < 0. 001 ), while the purified 2E8 mAb did not show any significant influences on the growth of the same cell line ( P > 0.05 ). Meanwhile, no significant inhibitory effects on the CD-19 K562 cells were identified in the 2E8-NCTD, 2E8 mAb, or control groups, indicating a significant targeting effect of 2E8-NCTD against Nalm-6 cells.Conclusions The immunotoxin 2E8-NCTD can be synthesized by activated ester method. It has target killing effects on CD+19 Nalm-6 leukemia cells in vitro.

OBJECTIVETo identify clinical and laboratory abnormalities and genetic defect of Fechtner syndrome in a Chinese family.

METHODSThe characteristic morphological features of platelets and leukocytes were examined on blood smears with Wright's-Giemsa staining and ultrastructure of platelet and leukocyte were investigated under electron microscope. Genomic DNA was isolated from peripheral blood of the proband and 9 members of his family. All the exons and exon-intron boundaries of the MYH9 gene were amplified by PCR followed by direct sequencing.

RESULTSPatients presented the characteristic clinical features including macrothrombocytopenia, leukocyte inclusions and/or hereditary nephritis. A heterozygous C to T mutation was found in the proband and three members of his family at nucleotide 5981 in exon 40 of MYH9 gene, resulting in a nonsense mutation which encoded truncated protein due to premature termination at the Arg 1933 codon.

CONCLUSIONIt is the first report of a Chinese family with Fechtner syndrome. The Arg (CGA) 1933--> stop (TGA) nonsense mutation in MYH9 gene is a causative genetic defect.

Adult ; Codon, Nonsense ; DNA Mutational Analysis ; Exons ; genetics ; Humans ; Inclusion Bodies ; genetics ; Male ; Molecular Motor Proteins ; genetics ; Myosin Heavy Chains ; genetics ; Nephritis, Hereditary ; genetics ; Pedigree ; Syndrome ; Thrombocytopenia ; genetics

To observe in vitro the effect of rat drug serum on the proliferation of HSC-T6 hepatic stellate cells in the pharmacokinetic model for determining peoniflorin in Fufang Biejia Ruangan tablet, in order to discover the rational daily administration frequency of Fufang Biejia Ruangan tablet. Fufang Biejia Ruangan tablet was orally administered to rats with different daily administration frequency. Their blood was collected from veins behind eye sockets at different time points before the administration and after the first administration, in order to determine the concentration of peoniflorin in blood plasma and the effect of rat drug serums on the proliferation of HSC-T6. A comprehensive analysis was made on the relationship between pharmacodynamics and pharmacokinetics to determine the rational daily administration frequency of Fufang Biejia Ruangan tablet. The results showed a good correlation between the inhibitory effect of Fufang Biejia Ruangan tablet-contained serum on HSC-T6 and the concentration of peoniflorin in blood. The two-time administration group showed higher pharmacologic and pharmacokinetic AUCs than one-time administration and three-time administration groups. In conclusion, Fufang Biejia Ruangan table is recommended to be taken twice a day for treating liver fibrosis in chronic hepatitis.
Administration, Oral ; Animals ; Area Under Curve ; Benzoates ; administration & dosage ; blood ; pharmacokinetics ; Bridged-Ring Compounds ; administration & dosage ; blood ; pharmacokinetics ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Hepatic Stellate Cells ; drug effects ; metabolism ; Liver Cirrhosis ; drug therapy ; metabolism ; Male ; Monoterpenes ; Rats ; Rats, Sprague-Dawley ; Tablets ; administration & dosage ; pharmacokinetics