OBJECTIVETo investigate the effects of Shadu Cao Mixture (SDCM, traditional Chinese medicine) on immune functions of immunosuppression mice.

METHODSFifty BALB/C mice were randomly divided into blank control group, model group, SDCM low-dose, middle-dose and high-dose group. Except the blank control group, other groups were intraperitoneal injected with cyclophosphamide (40 mg/kg) to establish immunosuppression mice model. The blank control group and model group received gavage administration with nonnal saline, while the other groups received gavage administration with different doses of SDCM (10, 20, 40 m/kg for 15 days) respectively. The number of leukocytes and serum levels of interleukin-2 (IL-2), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in peripheral blood, spleen index, and the function of NK cells were measured.

RESULTSCompared with the model group , SDCM increased the number of leukocytes and serum concentrations of IL-2, TNF-α and IFN-γ in peripheral blood and improved the spleen index and the function of NK cells significantly (P < 0.05-0.01).

CONCLUSIONSDCM could remarkably enhance the immune functions of immunosuppression mice induced by cyclophosphamide.

Animals ; Cyclophosphamide ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Immunosuppression ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Killer Cells, Natural ; immunology ; Mice ; Mice, Inbred BALB C ; Spleen ; immunology ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo look for the active fraction of ethanol extract of Genkwa Flos (EGF) induced hepatotoxicity and develop an UPLC fingerprint of the active fraction.

METHODTarget fraction of EGF induced hepatotoxicity was guided by the serum biochemical and histopathology methods. The UPLC method was applied to establish the chromatographic fingerprint. The separation was achieved on a BEH C18 column (2.1 mm x 50 mm, 1.7 microm) with a mobile phase consisting of acetonitrile and water containing 0.05% phosphate acid running gradient elution. The detection was carried out at 210 nm and the analysis was finished within 10 min.

RESULTThe chloroform phase of EGF could be responsible for the hepatotoxicity of this herb. The common mode of the UPLC fingerprint was set up under the established condition. There were 17 common peaks in fourteen batches of herbs, eight of which were identified, and the similar degrees of the fourteen batches to the common mode were between 0.890-0.999.

CONCLUSIONIt is easy to locate the chloroform extraction of EGF with hepatotoxicity. And the UPLC fingerprint was developed for the above fraction, which could provide valuable references for safe and effective clinical use of EGF.

Animals ; Asteraceae ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; toxicity ; Flowers ; chemistry ; Humans ; Liver ; drug effects ; Male ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effects of gestational isoflurane exposure on postnatal memory and learning and growth-associated protein-43 (GAP-43), neuropeptide Y(NPY) expression in the hippocampus of pups.

METHODSTwelve maternal Sprague-Dawley rats at gestation d 18(E18) were randomly divided into isoflurane group (n=6) and control group (n=6). Rats in isoflurane group were exposed to 1.3 % isoflurane for 6 h. For control group, animals breathed in 30 % oxygen and air mixed gas at the same condition. Spatial learning and memory of the offspring were determined with the Morris Water Maze(MWM) after postnatal 4 weeks. The changes of GAP-43 and NPY expression in the hippocampal CA1 region of the pups were determined by immunohistochemistry.

RESULTSIn MWM training, the escape latency to platform of the pups in isoflurane group was significantly longer, and the time spent in the third quadrant and times of original platform crossing were less than those of control animals (P<0.05). The number and optical density of GAP-43 and NPY positive neurons in the hippocampus of pups decreased significantly in the isoflurane group compared with the controls (P <0.01).

CONCLUSIONIsoflurane exposure in pregnant rats significantly impairs the spatial memory and learning of their pups at a juvenile age, which may be associated with the down-regulation of GAP-43 and NPY in the hippocampus.

Animals ; Female ; GAP-43 Protein ; metabolism ; Hippocampus ; drug effects ; metabolism ; Isoflurane ; pharmacology ; Maze Learning ; drug effects ; Neuropeptide Y ; metabolism ; Pregnancy ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Sprague-Dawley

BACKGROUNDPemetrexed is a novel folic acid antagonist with multiple targets, which has been widely used in the treatment of non-small cell lung cancer (NSCLC). The objective of this study was to compare the effects and toxicities in NSCLC patients treated with pemetrexed monotherapy versus pemetrexed plus a platinum combination agent, so as to provide a basis for standard second-line chemotherapy.

METHODSThe clinical data of 52 patients with NSCLC who were admitted to Shanghai Chest Hospital from August 2006 to October 2008 were retrospectively analyzed. Ten of the 52 patients received pemetrexed monotherapy, and the other 42 patients received the pemetrexed plus platinum regimen. The primary end point was overall survival (OS). The progression-free survival time (PFS) was analyzed and the effects and toxicities were assessed. Survival analysis was evaluated by Kaplan-Meier method. Single factor analysis and the COX regression model were done to analyze the relationship between the influential factors and the prognosis of disease. The elderly patients (> or = 60 years old) were analyzed separately as a subgroup.

RESULTSNo statistically significant increase in OS (chi(2) = 0.09, P = 0.76), PFS (chi(2) = 0.15, P = 0.70), disease control rate (DCR) (chi(2) = 0.06, P = 0.81) or 1-year survival rate (chi(2) = 0.33, P = 0.57) was found between the two regimens. Single factor analysis showed that the factors including surgery history, PS score before treatment, clinical stage, and response to second-line treatment influenced the prognosis of NSCLC (all P < 0.05). COX regression analysis demonstrated that surgery history (P = 0.041) and performance status (PS) score before treatment (P = 0.043) may be associated with survival. The toxicity of the two regimens was similar. In the subgroup of elderly patients, no significant difference in OS (chi(2) = 0.01, P = 0.94), PFS (chi(2) = 0.14, P = 0.70), DCR (chi(2) = 0.004, P = 0.95), or 1-year survival rate (chi(2) = 0.03, P = 0.87) was found between the two regimens. The toxicity of combination therapy was significantly higher in terms of hematologic (chi(2) = 9.95, P = 0.01) and gastrointestinal adverse events (chi(2) = 7.66, P = 0.03).

CONCLUSIONSThere is no significant difference in survival or side effects between these two regimens. For elderly patients (> or = 60), pemetrexed monotherapy shows similar efficacy and a better safety profile when compared with pemetrexed combination therapy.

Aged ; Aged, 80 and over ; Antineoplastic Agents ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Female ; Glutamates ; therapeutic use ; Guanine ; analogs & derivatives ; therapeutic use ; Humans ; Lung Neoplasms ; drug therapy ; Male ; Middle Aged ; Pemetrexed ; Platinum ; therapeutic use ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate the effects of the epidermal growth factor on the mRNA expression of endothelin-1 and its receptors (ET(A)R, ET(B)R) in hormone refractory prostate cancer (HRPC) PC-3 cell lines.

METHODSPC-3 cells were cultured in vitro. After the treatment with EGF, the mRNA expressions of endothelin-1, ET(A)R and ET(B)R were detected by RT-PCR in PC-3 cell lines. The levels of the mRNA expression of endothelin-1 and its receptors were examined at different time points by RT-PCR.

RESULTSThe expressions of endothelin-1 and ET(A)R mRNA but not the mRNA expression of ET(B)R was observed in PC-3 cell lines. After 24 hours of treatment with EGF, the expressions of endothelin-1 and ET(A)R in PC-3 cell lines were both up-regulated and there was significant difference (P < 0.05) between the experimental and control groups. Different expression levels of endothelin-1 and ET(A)R mRNA were noted at different time points of EGF intervention, up-regulated with the increase of treatment time, and with significant difference (P < 0.05).

CONCLUSIONEGF can up-regulate the mRNA expressions of endothelin-1 and ET(A)R in PC-3 cell lines and play a great role in prostate cancer progression, which may offer a substructure of molecular biology for the treatment of HRPC.

Cell Line, Tumor ; Endothelin-1 ; genetics ; Epidermal Growth Factor ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Male ; Prostatic Neoplasms ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism ; Receptor, Endothelin A ; genetics ; Receptor, Endothelin B ; genetics ; Receptors, Endothelin ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVEThe prostate apoptosis response factor-4 (Par-4) gene was originally identified by differential screening for genes that are up-regulated when prostate cells are induced to undergo apoptosis. Par-4 was found to possess potent apoptotic activity in various cellular systems in response to numerous stimuli. The aim of this study was to explore the effects of small interfering RNA (siRNA) against Par-4 gene on the apoptosis of human bone marrow mesenchymal stem cells (hBMSCs) exposed to glutamate.

METHODSPrimary culture of hBMSCs was carried out and siRNAs targeted Par-4 gene (Par-4-SiRNA) were chemically synthesized. Eukaryocytic expression vector was built and were transfected into hBMSCs with liposome. After selecting with G418, the stable cell clones were treated with glutamate. The expression of Par-4 mRNA was determined by real-time PCR. The apoptosis of hBMSCs was quantified by flow cytometry. Western blotting was used to detect the protein levels of phosphorylated Akt1 (Thr308). Relative Caspase-3 activity was determined by colorimetric assay.

RESULTSThe Par-4-SiRNA-1 and Par-4-siRNA-2 could markedly down-regulate the mRNA levels of Par-4 gene in hBMSCs. With the transfections of Par-4-SiRNA-1 and Par-4-SiRNA-2, the levels of Par-4 mRNA were respectively decreased by 88% and 67%. Both Par-4-SiRNA-1 and Par-4-SiRNA-2 inhibited significantly the apoptosis of hBMSCs induced by glutamate, in which the percentages of apoptotic cells were respectively decreased to 38.80% +/- 3.97% (P < 0.01) and 45.49% +/- 4.32% (P < 0.01) from 60.30% +/- 6.82%. Western blot assays demonstrated that, glutamate down-regulated the expression of phosphorylated Akt1 proteins in hBMSCs (89.07 +/- 6.42 and 28.30 +/- 5.65, respectively, P < 0.01). However, Par-4-SiRNA-1 and Par-4-SiRNA-2 could markedly recover the down-regulation of Akt1 proteins induced by glutamate (63.56 +/- 6.75 and 45.59 +/- 4.88, respectively, P < 0.01). And the relative Caspase-3 activity which was enhanced by the treatment with glutamate (0.1428 +/- 0.0495 and 0.8616 +/- 0.1051, P < 0.01), was suppressed by Par-4-SiRNA-1 and Par-4-SiRNA-2 (0.8616 +/- 0.1051 and 0.6581 +/- 0.0555, respectively, P < 0.01).

CONCLUSIONSiRNA against Par-4 gene could inhibit the apoptosis of hBMSCs induced by glutamate, and its inhibitory effects may be mediated by the up-regulation of phosphorylated Akt1 and the suppression of the relative Caspase-3 activity.

Apoptosis ; genetics ; Apoptosis Regulatory Proteins ; genetics ; Bone Marrow Cells ; cytology ; metabolism ; Caspase 3 ; metabolism ; Cells, Cultured ; Gene Expression Regulation ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Small Interfering

BACKGROUND: Until now, there are no reliable methods for the treatment of urinary incontinence after radical prostatectomy. Some limitations exist in drug therapy, mid-urethral suspension, and filling agent treatment. Therefore, the use of autologous adipose-derived stem cells (ADSCs) is expected to become a first-line treatment strategy for urinary incontinence after radical prostatectomy. OBJECTIVE: To report our initial experience with transurethral injection of autologous ADSCs for the treatment of urinary incontinence after radical prostatectomy. METHODS: Patients and their families were informed of possible risks and benefits prior to the participation in the trial. After providing written informed consent, six patients with persistent urinary incontinence after radical prostatectomy were enrolled in the study. Under general anesthesia, about 50 mL of adipose tissue was obtained from each patient by liposuction. ADSCs were obtained by separation with centrifugation using the Celution cell-processing device. A mixture of ADSCs and adipose tissue was transurethrally injected into the submucosal space of the membranous urethra. Functional and anatomical improvement was assessed through a 24-hour pad test, validated patient questionnaire, urethral pressure profile, and magnetic resonance imaging (MRI) through 12-week follow-up. RESULTS AND CONCLUSION: Urine leakage volume was improved with time in all patients in the 24-hour pad test, with the exemption of temporal deterioration in two patients at the first 2 weeks post-injection. Subjective symptoms and quality of life assessed on the basis of questionnaire results showed similar improvement. The mean maximum urethral closing pressure increased from 4.312 kPa to 6.223 kPa at 12 weeks after cell injection. MRI results showed an increase in functional profile length (from 6.1 to 8.3 mm) between the lower rim of the pubic bone and the bladder neck. Adverse events, such as pelvic pain, inflammation, or de novo urgency, were undetected in any case during the follow-up. To conclude, the transurethral injection of autologous ADSCs can be a safe and effective treatment for urinary incontinence after radical prostatectomy.

Objective:Due to the limitation of traditional identification methods of Chinese medicinal materials， the study established a rapid method to identify Persicae Semen mixed with Armeniacae Semen Amarum by allele-specific polymerase chain reaction （PCR）. Method:By comparing the ribosomal DNA internal transcribed spacer （ITS） gene sequences of Persicae Semen and Armeniacae Semen Amarum， single nucleotide polymorphism （SNP） sites were searched and specific primers were designed. Different Persicae Semen and Armeniacae Semen Amarum samples were amplified by PCR， the effects of annealing temperature， primer concentration and cycle number on the PCR reaction system were optimized， and the specificity and detection limit of this method were investigated. In addition， the established PCR method was used to detect the samples of Persicae Semen mixed with different proportion of Armeniacae Semen Amarum from different sources and producing areas. Result:A specific PCR method for identifying Persicae Semen mixed with Armeniacae Semen Amarum was established. When the annealing temperature was 63 ℃ and the number of primer cycles was 30， only Armeniacae Semen Amarum could be amplified with 432 bp specific band， while Persicae Semen samples did not have this band. The minimum detection limit of this method for Armeniacae Semen Amarum was 0.2 ng， and the detection limit for Armeniacae Semen Amarum adulterated in Persicae Semen was 1%. Conclusion:The established allele-specific PCR method can accurately detect whether there is Armeniacae Semen Amarum in Persicae Semen， which can provide experimental basis for the quality control of Persicae Semen and guarantee the safety of its clinical use.

Purpose of this study was to analyse the characteristics of clinical, iconographical, pathological and treatment methods of Langerhans cell histiocytosis (LCH), so as to improve the diagnosis and treatment level of this disease. The clinical data of 35 LCH patients were studied retrospectively. These patients were divided into 2 groups according to age <14 years old and ≥ 14 years old. The clinical symptoms were analysed and the signs, imageology and pathology manifestation and treatment results were evaluated. The results showed that LCH clinical manifestations were diverse and complex. Surgical treatment for patients with single system involvement of LCH was better than that of multi-system involvement of LCH (MS-LCH). For the latter, combined chemotherapy effects was better. After 3-year follow-up, 1-year OS was 94% ± 4%, 2-years OS was 91% ± 5%, 3-year OS was 86% ± 7%. 3 years OS of group <14 years old and ≥ 14 years old was 94% ± 6% and 81% ± 10% respectively. The OS of former was better than that of the later, but because a small number of cases, this difference was not statistically significant. It is concluded that LCH is easy to be misdiagnosed, the pathological biopsy is the gold standard of LCH diagnosis. The PET-CT can be of great help in identifying stages and finding lesion areas of the disease. Pulmonary Langerhans cell histiocytosis (PLCH) is more common in adult. Combined chemotherapy can improve the prognosis of the patients. The treatment methods should be choosed according to the stage and classification of disease.
Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Histiocytosis, Langerhans-Cell ; diagnosis ; drug therapy ; pathology ; Humans ; Infant ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Young Adult