Animals ; Anti-HIV Agents ; pharmacology ; HIV ; drug effects ; genetics ; physiology ; HIV Infections ; drug therapy ; immunology ; virology ; Humans ; Mucous Membrane ; immunology ; virology ; Virus Replication ; drug effects

BACKGROUND:Large-scale expansion of undifferentiated and multipotential adipose-derived stem cells using serum-free culture system is a difficult issue to be resolved. OBJECTIVE:To establish an in vitro culture system combined with the extracellular matrix in order to investigate the efficiency, effectiveness and security of extracellular matrix on expanding adipose-derived stem cells. METHODS:In vitro isolated adipose-derived stem cells were seeded in traditional two-dimensional plastic plates and extracellular matrix-coated plates supplemented with serum-free medium respectively. After in vitro expansion, total cellnumber, expression of cellsurface markers, cellsenescence degree and multipotent differentiation ability (adipogenic, osteoblastic and chondrogenic differentiation) of adipose-derived stem cells cultured under both conditions were detected and compared. Moreover, the clinical safety of adipose-derived stem cells expanded in extracellular matrix-coated plates was investigated. RESULTS AND CONCLUSION:Total cellnumber of passage 5 adipose-derived stem cells cultured in extracellular matrix-coated plates was 10 times more than that in traditional two-dimensional plastic plates. Flow-cytometric analysis showed that adipose-derived stem cells cultured with extracellular matrix expressed stem cellsurface markers. cellular senescence examination showed that almost al of passage 15 adipose-derived stem cells cultured with extracellular matrix showed no aging, while most passage 5 adipose-derived stem cells cultured by the two-dimensional system aged and lost their proliferation ability. Multidirectional induction of adipose-derived stem cells showed that passage 15 adipose-derived stem cells cultured with extracellular matrix could stil differentiate into adipocytes, osteoblasts and chondrocytes as passage 5 adipose-derived stem cells did, which performed much better than the induced differentiations of passage 5 adipose-derived stem cells cultured by the two-dimensional system. Karyotype analysis and in vivo invasion experiment insured the clinical safety of adipose-derived stem cells expanded with extracellular matrix. Al above results suggest a safe and more efficient expansion system of extracellular matrix for clinical application using the serum-free culture system combined with extracellular matrix.

Object To establish the method for quality control of Fushengkang Capsule. Methods Semen Livistonae Chinensis, Fructus Camptothecae Acuminatae and Rhizoma Curcumae in this capsule were identified by TLC. The content of camptothecine was determined by HPLC. Condition of HPLC was: Dikma Diamonsil C 18 column (250 mm? 4 6 mm, 5 ?m) the mobile phase was a mixture of methanol water (55∶45), the flow rate was 0 8 mL/min, and detection wavelength at 254 nm. Results The spots of TLC were clear and concentrative. There was a good linear relationship for camptothecine within the range of 0 050 6 - 0 253 0 ?g. The average recovery was 96 90%, RSD=1 74% (n=6). Conclusion This method is sensitive, accurate, reproducible and exclusive. It can be used for quality control of Fushengkang Capsule

Objective To explore the interference effect for the compliance of attribution training to the patients suffered from the kidney complex in hormone treatment.Methods 256 cases of patients suffered from kidney complex are divided into 130 as a training group and 126 cases as a comparison group randomly.The training group was given by the normal nursing,hormone medicine treatment and also the interference of attribution training nursing measurements;the comparison group was only given the normal nursing,hormone medicine treatment.Two groups of the patients were assessed by the psychology quality of the emotion application measure meter to the patients after the treatment of 6 months separately;The Morisky-Green measure meter(MG)was used for the assessment of the hormone treatment compliance of two groups of the patients.Results Various items of assessments of the psychology quality of training groups of patients are better than the comparison group evidently after the treatment of 6 months separately,there were significant differences between the two groups(t=3.05,3.37,3.72,P<0.01);The compliance percentage of the hormone treatment of training group is 80.77% but the compliance percentage of the hormone treatment of comparison group is 62.70%,there were significant differences between the two groups(χ2=10.34,P<0.01).Conclusions The attribution training could impreve the compliance of the patients suffered from kidney complex in the psychology quality and hormone treatment 80 as to effectively reduce recrudescing and improve the living quality of the patients.

BACKGROUNDRecently, the number of patients with prostate cancer who needed to be treated with radical prostatectomy increased rapidly in China. There is still a difference between clinical staging and the post-operative final pathologic staging; hence, an excellent tool for accurately predicting the pathologic stages of prostate cancer is needed urgently in clinical practice. The Partin tables are the most popular and widely used tool for predicting the pathologic stages of prostate cancer because of its high accuracy and ease of implementation. The aim of this study was to externally validate the accuracy of the three versions of the Partin tables in predicting the post-operative pathologic stages in Chinese patients with prostate cancer.

METHODSWe retrospectively analyzed the data from 203 patients with prostate cancer who underwent radical prostatectomies between June 2000 and May 2012. The accuracies of the three versions of the Partin tables in predicting the post-operative pathologic stages in Chinese patients with prostate cancer were evaluated using the area under the receiver operating characteristic curve (AUC).

RESULTSUsing the 1997, 2001, and 2007 Partin tables for predicting the current cases, the AUC of organ confinement (OC) was 0.877, 0.788, and 0.726; the AUC of extracapsular extension (ECE) was 0.525, 0.615, and 0.608; the AUC of seminal vesicle invasion (SVI) was 0.875, 0.649, and 0.820; and the AUC of pelvic lymph node invasion (LNI) was 0.808, 0.758, and 0.735 respectively.

CONCLUSIONSThe accuracies of the three versions of Partin tables in predicting OC, SVI, and LNI were good, especially the 2001 Partin table for SVI. In contrast, the accuracy of the three versions of the Partin tables in predicting ECE was fair. The 1997 Partin table was much better than the 2007 table in predicting OC, and the 2001 table in predicting SVI. The 2007 Partin table did not show any advantages.

Aged ; Humans ; Male ; Middle Aged ; Neoplasm Grading ; Neoplasm Staging ; Nomograms ; Prostate-Specific Antigen ; blood ; Prostatectomy ; Prostatic Neoplasms ; pathology ; surgery ; Retrospective Studies

AIMTo manage male infertility with obstructive azoospermia by means of percutaneous epididymal sperm aspiration (PESA) and intrauterine insemination (IUI).

METHODSNinety azoospermic patients with congenital bilateral absence of the vas deferens (BAVD, n=58) or bilateral caudal epididymal obstruction (BCEO, n=32) requesting for fine needle aspiration (FNA), PESA and IUI were recruited. The obstruction was diagnosed by vasography and determination of the fructose, carnitine and alpha-glucosidase levels in the seminal fluid.

RESULTSThe mean sperm motility, density, abnormal sperm and total sperm count of the caput epdidymis were 16 %+/-22 %, (12+/-31) x 10(6)/mL, 55 %+/-36 % and (16+/-14) x 10(6), respectively. In the 90 couples, a total of 74 PESA procedures and 66 cycles of IUI were performed. Three pregnancies resulted, including one twin pregnancy giving birth to two healthy boys, one single pregnancy with a healthy girl and another single pregnancy aborted at week 6 of conception. The pregnancy rate per IUI cycle was 4.5 %.

CONCLUSIONThe birth of normal, healthy infants by IUI using PESA indicates that the caput epididymal sperm possess fertilization capacity. The PESA-IUI programme is a practical and economical procedure for the management of patients with obstructive azoospermia.

Adult ; Biopsy, Needle ; Epididymis ; cytology ; Female ; Humans ; Insemination, Artificial ; methods ; Male ; Oligospermia ; therapy ; Pregnancy ; Pregnancy Outcome ; Spermatozoa ; cytology

Objective To investigate the mRNA and protein expressions of high mobility group box-1 ( HMGB1 ) in the tumor tissues and sera of patients with laryngeal squamous cell carcinoma (LSCC) and their clinical significance. Methods Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot were used to detected the expressions of HMGB1 mRNA and protein in the tumors and adjacent normal epithelial tissues in 30 patients with LSCC.Serum HMGB1 protein levels in the patients with LSCC and in 10 healthy volunteers were detected with enzyme-linked immunosorbent adsorption experiment( ELISA ).Results RT-PCR demonstrated that the mean relative mRNA expression levels of HMGB1 (HMGB1/GAPDH) in LSCC tissues and in adjacent normal epithelial tissues were 1.25 ±0.12 and 0.32 ± 0.04,respectively ( t =40.27,P ＜ 0.05 ).Western blot revealed that the mean relative protein expression levels of H MGB 1 ( HMGB1/β-actin) were 1.29 ± 0.10 and 0.34 ± 0.03 (t =49.84,P ＜ 0.05 ),respectively.Both mRNA and protein expression levels of HMGB1 were associated with T stage,clinical stage,lymph node metastasis status and smoking ( all P ＜ 0.05 ),but no significant correlation with age,alcohol consumption and primary tumor grade and location (all P ＞0.05 ).Mean serum HMGB1 protein levels in patients with LSCC and healthy volunteers were (24.80 ± 14.08) ng/ml and (23.58 ± 14.69) ng/ml ( t =0.37,P ＞ 0.05 ).Conclusions Both mRNA and protein expressions of HMGB1 were obviously elevated in LSCC,which were associated closely with T stage,clinical stage and lymph node metastasis.

Objective To study the effect of gene optimization on the expression and purification of HDV small antigen produced by genetic engineering.Methods Based on the colon preference of E.coli,the HDV small antigen original gene from GenBank was optimized.Both the original gene and the optimized gene expressed in prokaryotic cells,SDS-PAGE was made to analyze the protein expression yield and to decide which protein expression style was more proportion than the other.Furthermore,two antigens were purified by chromatography in order to compare the purity by SDS-PAGE and Image Lab software.Results SDSPAGE indicated that the molecular weight of target proteins from two groups were the same as we expected.Gene optimization resulted in the higher yield and it could make the product more soluble.After chromatography,the purity of target protein from optimized gene was up to 96.3％,obviously purer than that from original gene.Conclusion Gene optimization could increase the protein expression yield and solubility of genetic engineering HDV small antigen.In addition,the product from the optimized gene group was easier to be purified for diagnosis usage.

To investigate the effect of Yunkang Oral Liquid on preventing lipopolysaccharide(LPS)-induced abortion and regulating immune tolerance in mice,sixty normal ICR mice were randomly divided into normal group,model group,Yunkang Oral Liquid high,middle and low dose groups and progesterone group.Abortion model was established by tail vain injection of LPS(0.1μg/mouse)on the 7th day of pregnancy.Since the first day of pregnancy,the same volume of distilled water,Yunkang Oral Liquid at the dose of 36,18 and 9 m L·kg~(-1)·d~(-1),and progesterone at the dose of 0.038 g·kg~(-1)·d~(-1)were given in corresponding groups.The mice were sacrificed at the 9th day of pregnancy,and the embryo loss of each group was calculated.The levels of Th1 type cytokines(TNF-α,IFN-γ)and Th2 type cytokines(IL-4,IL-10)in uterus homogenate were detected by ELISA.HE staining was performed to examine the histopathological changes in the decidua.The expression levels of CD14,F4/80 in macrophages of uterus were detected by immunohistochemistry.Western blot was used to investigate the protein expression of TLR4,MyD88 and NF-κB in uterine decidua.In our study,all Yunkang Oral Liquid groups could significantly reduce the embryo absorption rate of mice,while high dose group can significantly increase the levels of IL-10 and IL-4;both medium and high dose groups can significantly decrease TNF-α,and IFN-γlevelsin the uterus of model mice,reduce the protein expression of NF-κB,MyD88 and TLR4 in uterine decidua tissue.Various treatment groups could reduce the counts of F4/80,CD14 macrophages and decrease expression area in uterine tissue.Our results showed that Yunkang Oral Liquid could prevent LPS-induced abortion,and the mechanism may be associated with inhibiting the TLR4/MyD88/NF-κB signaling pathway and regulating the balance of Th1/Th2 immune factors,which could improve the endometrial receptivity of mice,and promote the development of decidua and implantation of embryo.
Abortion, Induced ; Animals ; Female ; Immune Tolerance ; Lipopolysaccharides ; Mice ; Mice, Inbred ICR ; NF-kappa B ; Pregnancy

OBJECTIVETo study the clinical and epidemiologic characteristics and the serum types of enterovirus of hand, foot and mouth disease (HFMD) in children.

METHODSThe RT-nPCR method was established with universal primers within 5' untranslated region of enterovirus and VP1 region of Coxsackievirus A16 (CAV16) and enterovirus 71 (EV 71). Enteroviruses were detected with RT-nPCR in 237 children with HFMD. Clinical and epidemiologic characteristics and serum types of enterovirus of the patients with HFMD were studied.

RESULTSThe patients'age ranged from 7 months to 11 years (mean 4.2 +/- 0.5 years). The majority (94.5%) were less than 6 years old. HFMD was mostly seen in spring and winter (67.9%). Oral mucosal pox or ulcer as well as hand and foot rashes were observed in all 237 patients. Fever occurred in 141 patients (59.5%). Of the 237 patients, 133 (56.1%) were RT-nPCR positive. Of the 133 cases, 38 were positive for EV71, 64 were positive for CAV16, and 31 were negative for both EV71 and CAV16. The patients infected by different types of enteroviruses had similar clinical characteristics. Gene colon and sequence analysis for 12 strains of enteroviruses PCR positive products presented as EV71 (n=5), CAV16 (n=5), ECHO13 (n=1), and CAV5 (n=1).

CONCLUSIONSHFMD tends to occur in younger children less than 6 years old. The majority are affected in spring and winter. EV71 and CAV16 are common pathogens of HFMD. There is no relationship between clinical characteristics and serum types of enteroviruses in HFMD patients.

Child ; Child, Preschool ; Enterovirus ; classification ; Female ; Hand, Foot and Mouth Disease ; virology ; Humans ; Infant ; Male ; Reverse Transcriptase Polymerase Chain Reaction ; Seasons ; Serotyping