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Objective: To evaluate the clinical effect and safety of human umbilical cord mesenchymal stem cells (HUCMSCs) in the treatment of lung injury caused by acute paraquat poisoning., Methods: Thirteen patients with lung injury caused by acute paraquat poisoning, who were admitted to Guangzhou No. 12 People's Hospital from December 2008 to December 2012, were divided into HUCMSC group (n = 5) and control group (n = 8). All patients received conventional treatment, while the HUCMSC group was treated with HUCMSCs as an addition. Sequential Organ Failure Assessment (SOFA) system, which was created by the Infection Section of European Society of Intensive Care Medicine, and Acute Physiology and Chronic Health Evaluation II were used to acquire the SOFA scores of patients. The lung injury was evaluated with lung injury score (LIS). The two groups were compared with respect to maximum SOFA scores at 1, 3, 5, 7, 14, and 15 days after paraquat poisoning., Results: The HUCMSC group showed significantly lower maximum SOFA scores than the control group at 15d after poisoning (1.80 ± 2.05 vs 13.50 ± 7.59, P < 0.05). The LISs of the HUCMSC group after treatment (0.45 ± 0.27) were significantly lower than those of the HUCMSC group before treatment (1.15 ± 0.34) and those of the control group after treatment (2.94 ± 1.20) (P < 0.01). In the HUCMSC group, all patients survived, and they complained no discomfort and showed normal liver, kidney, and lung functions in reexamination; one patient showed incompletely absorbed shadow in the posterior segment of the left lower lobe of the lung during lung CT scan, and no abnormal findings were seen in other patients. In the control group, one patient survived, and others died. No adverse reactions, such as chill and fever, were presented in the HUCMSC group., Conclusion: HUCMSCs show promise for clinical application in the treatment of lung injury caused by acute paraquat poisoning.

Objective: To compare the difference of effects on SiO(2)-induced alveolitis and early fibrosis between bone marrow-derived mesenchymal-like stem cells (BM-MSCs) and BM-MSCs transfected by pcDNA3.1-HGF and to explore the mechanism of this effects., Methods: The Primary BM-MSCs from Wistar male young rats were cultured and labeled by 4, 6-diamidino-2-phenylindole (DAPI). Fifty Wistar rats were randomly divided into 3 groups:model group (10 rats),which was administered with SiO(2) by the trache, the next day,injected PBS via the tail vein; BM-MSCs group (20 rats),which was administered with SiO(2) by the trache, the next day,injected with 1 ml suspension of BM-MSCs via the tail vein; pcDNA3.1-HGF plus BM-MSC group (20 rats),which was administered with SiO(2) by the trache, the next day,injected with 1 ml suspension of BM-MSCs transfected by pcDNA3.1-HGF via the tail vein. On the 14th and 28th days after treatment, half of the animals were sacrificed, respectively, and the lungs were harvested for frozen section to observe the cell marked by DAPI. HE staining under a fluorescent microscope, and to observe the pulmonary alveolitis and fibrosis by HE and Masson staining under a light microscope. Western blot assay was used to detect the expression of HGF in rat lungs. The expression levels of tumor necrosis factor-α (TNF-α) in pulmonary tissues were analyzed quantitatively by ELISA. The contents of HYP in pulmonary tissues were analyzed quantitatively by sample hydrolysis method., Results: On the 14th and 28th days after treatment, the scores of pulmonary alveolitis and early fibrosis in pcDNA3.1-HGF plus BM-MSCs group were 2.36 ± 0.17, 2.8 ± 0.14 and 0.1 ± 0.11, 1.16 ± 0.13, which were significantly lower than those (1.68 ± 0.17, 1.58 ± 0.31 and 0.54 ± 0.15, 1.36 ± 0.13) in BM-MSCs group, also which were significantly lower those (2.36 ± 0.17, 2.80 ± 0.14 and 0.64 ± 0.09, 1.84 ± 0.17) in model group (P < 0.05); On the 14th and 28th days after treatment, the TNF-α contents of pulmonary tissues in pcDNA3.1-HGF plus BM-MSCs group were 280.4 ± 23.11 and 249.78 ± 22.33 pg/mg, which were significantly lower than those (341.58 ± 35.34, 442.29 ± 36.76 pg/mg and 319.51 ± 17.84, 348.53 ± 33.95 pg/mg) in BM-MSCs and model groups (P < 0.05); On the 14th and 28th days after treatment, the HYP contents of pulmonary tissues in pcDNA3.1-HGF plus BM-MSCs group were 0.46 ± 0.04 and 0.65 ± 0.05 µg/mg, which were significantly lower than those (0.63 ± 0.04, 1.04 ± 0.07 µg/mg and 0.72 ± 0.60, 1.39 ± 0.60 µg/mg) in BM-MSCs and model groups (P < 0.05)., Conclusion: The effects of BM-MSCs transfected by pcDNA3.1-HGF on suppressing pulmonary alveolitis and early fibrosis induced by SiO2 were better than those of BM-MSCs. The mechanism may be associated with the reduced pulmonary inflammation.

Objective: To compare the pulmonary alveolitis and the early fibrosis of pulmonary fibrosis induced by quartz dust and bleomycin in rats, and investigate their mechanism., Methods: The female rats were divided into three groups: control group exposed to normal saline by the trachea; SiO2 group exposed to SiO2 by the trachea; BLM group exposed to BLM A5 by the trachea. Each half of the animals were sacrificed on the 7th and 14th day after exposure. The lungs of rats were collected to observe pulmonary alveolitis by HE staining and to observe fibrosis by saturated picric acid sirius red staining. The expression of tumor necrosis factor-alpha (TNF-alpha) and CD68 in pulmonary tissues were analyzed quantitatively by immunohistochemistry and image analysis system., Results: (1) The alveolitis and pulmonary fibrosis of rats in both SiO2 group and BLM group were became more serious gradually over time, HE staining under light microscope showed that BLM group on the 7th day had the most obvious alveolitis (2.814 +/- 0.832), the saturated picric acid sirius red staining under polarized light showed that BLM group on the 14th day had the worst pulmonary fibrosis (1284.57 +/- 554.72), which were significantly higher than those (103.69 +/- 18.29 and 111.78 +/- 37.45) in control group and SiO2 group on the 7th day (P < 0.05). (2) The results of immunohistochemistry examination indicated that the expression (17.100 +/- 1.831) of TNF-alpha in the BLM group on the 7th day was significantly higher than those (0.451 +/- 0.441, 7.909 +/- 1.275 and 13.506 +/- 1.454) in control group, SiO2 group on 7th day and BLM group on 14th day (P < 0.05). The expression (22.778 +/- 2.512) of TNF-alpha in the SiO2 group on the 14th day was significantly higher than those in control group, SiO2 group on 7th day and BLM group on 14th day (P < 0.05). The expression (134.941 +/- 35.951) of CD68 in the SiO2 group on the 14th day was significantly higher than those in control group, SiO2 group on 7th day and BLM group on 14th day (P < 0.05)., Conclusion: The early alveolitis of BLM-induced lung injury model was more serious than that of SiO2-induced lung injury model, and the fibrosis process of BLM-induced lung injury model was earlier than that of SiO2-induced lung injury model. TNF-alpha plays an important role in the course of both models, but macrophages is involved in SiO2-induced pulmonary in a more continuous way than in BLM-induced pulmonary fibrosis.

Objective: To evaluate the potential role of hepatocyte growth factor (HGF) combined with bone marrow mesenchymal stem cells (BMSC) autograft for the treatment of silicosis., Methods: Bone marrow (100 ml) was aspirated from a severe silicosis patient. BMSCs isolated, purified and cultured in vitro. When BMSC came to 70% confluence at passage 3, the culture medium was added liposomes (lipo2000) and plasmid-HGF (p-HGF) and cultured for 2 d. HGF-MSCSs (5 × 10(7) cells) were resuspended in 50 ml 0.9% sodium chloride (NS) and infused Intravenous drip at 3 consecutive times (once a week). Clinical follow-up were performed before and after treatment: (1) pulmonary high-kV X-ray, chest CT examination; (2) pulmonary function test; (3) determination of serum ceruloplasmin., Results: The symptoms such as coughing, chest tightness disappeared at 12 months after treatment. Pulmonary function tests showed significant changes after treatment: forced vital capacity (FVC) increased from 64.6% to 81.0%, forced expiratory volume in one second (FEV(1.0)) increased from 68.7% to 90.1%, 1 second rate (FEV(1.0)/FVC%) reduced from 111.6% to 107.1%, the maximum mid-expiratory flow (FEF(25%∼75%) decreased from 100.2% to 94.6%, forced expiratory vital capacity 75% of the moment bit of gas flow (MEF(75%)) increased from 99.2% to 113.5%, forced expiratory vital capacity 50% of the moment bit of gas flow (MEF(50%)) increased from 125.3% to 130.2%, forced expiratory vital capacity 25% of the moment bit of gas flow (MEF(25%)) reduced from 86.9% to 71.7%; serum ceruloplasmin levels decreased from 690 mg/L to 180.6 mg/L; lung high-kV X-ray at 1st review showed that diffuse lung nodules had been absorbed and getting smaller than before treatment; chest CT showed that the distribution and number of small nodules at double lung fields decreased than before treatment., Conclusion: HGF combined with BMSC transplantation may have some potential role for the treatment of silicosis patients.

Objective: To investigate the effect of Shenqi Fuzheng Injection (SFI) for hemopoietic and immune function reconstruction in patients with hematologic malignancies (HM) after chemotherapy., Methods: Eighty HM patients at remission inducing stage of initial treatment were randomly assigned to two groups, the treatment group treated with SFI (250 mL daily) plus chemotherapy and the control group only treated with chemotherapy for 14 days, with same supportive treatments administered to both. Levels of blood routine test, T lymphocyte subsets (CD3+, CD4+, CD4+ /CD8+) and B lymphocyte subsets CD3- CD19+ were determined before and after treatment, and the remission rate was assessed after treatment., Results: The remission rates in the two groups showed no significant difference [90% (36/40) vs 82.5% (33/40), P > 0.05] statistically. Levels of peripheral leucocyte count and hemoglobin as well as levels of CD3+, CD4+, CD4+ /CD8+; were significantly higher in the treatment group than in the control group (P < 0.05), but no significant difference was shown between groups in CD3- CD19+ level., Conclusion: SFI can lighten the inhibition of chemotherapy on hemopoietic function of bone marrow, and promote its recovery, enhance the immune function, and improve the quality of life in patients with HM undergoing chemotherapy.

Objective: To investigate the expression and significance of peroxisome proliferators-activated receptor gamma (PPAR gamma) in human glioma., Methods: Immunohistochemical staining for PPAR gamma was performed using biopsy specimens of human glioma of various histological types. Expression of PPAR gamma and GFAP in glioma cell lines SWO-38, U251 and SHG-44 were analyzed using Western blotting and reverse transcription-polymerase chain reaction (RT-PCR)., Results: Immunohistochemical study showed that PPAR gamma was expressed in glioma tissues with positive rate of 37.5%. Western blotting and RT-PCR showed that PPAR gamma was expressed in both glioma cell lines SWO-38 and U251, but not in SHG-44 cells. However, high expression of GFAP was detected in SHG-44 cells., Conclusion: PPAR gamma is associated with carcinogens of glioma. Actived PPAR gamma by agonist may be a novel approach to the treatment of glioma.

Objective: To observe the effects of exogenous human chorionic gonadotropin (hCG) on nude mice bearing transplanted endometrial carcinoma., Methods: Human endometrial carcinoma xenograft was transplanted in nude mice, and the effects of hCG injection on the tumor growth was evaluated according to tumorigenesis and xenograft weights. The expression of Ki-67 in the tumor was determined by immunohistochemistry, and HE staining was performed for morphological observation and measurement of the necrosis area in the tumor. The effect of hCG on fibrosis in the tumor was evaluated with Masson staining., Results: Compared to normal saline-treated tumor-bearing mice, the mice with hCG treatment showed increased tumor weight. HE staining for tumors in HCG-treatment group visualized tumor cell arrangement in glandular structure with smaller necrosis area, and Masson staining identified thick and compact collagen fibers as compared with the thin and loosely arranged fibers in saline-treated group. No significant difference was found in the Ki-67 expression in the tumors between the two groups., Conclusion: Exogenous hCG can promote the differentiation of the endometrial carcinoma cells in vivo.

Objective: To investigate the biological effects of PTEN gene on the malignant glioma cell line SHG-44. Firstly, A recombinant eukaryotic expression plasmid containing PTEN gene fused with EGFP (enhanced green fluorescence protein) gene was constructed. Secondly, the expression of the recombinant vector in human glioma cells was detected., Methods: (1) The human PTEN gene was amplified by RT-PCR and inserted into pEGFP-N1 that was selected by T-A subclone, and the recombinant expression vector was obtained. After the recombinant plasmids were transfected into glioma SHG-44 cells by cation polymex, expression of fusion protein was tested. (2) The stable transfected cells were selected by G418 and amplified. Light microscopy and growth curve were used to measure the effects of PTEN expression on cell morphology and proliferation. Expression of GFAP (glial fibillary acidic protein) was detected immunohistochemically., Results: (1) The positive recombinant was sequenced and demonstrated to have the same sequence as that of PTEN gene in GenBank. It was proved that the eukaryotic expression vector pEGFP-PTEN have been constructed successfully and expressed in SHG-44 cells. (2) The SHG-44 cell growth was changed obviously. The expression of GFAP was increased., Conclusion: The construction of PTEN eukaryotic expression vector containing green fluorescence protein gene will lay the basis for carrying out further studies on the function of PTEN gene.

Background & Objective: The prognosis of T-cell non-Hodgkin's lymphoma (T-NHL) is poor. Overexpression of myeloid cell leukemia-1 (Mcl-1) gene could inhibit irradiation-and drug-induced apoptosis in several lymphoma cell lines. This study was to detect the expression of Mcl-1 in T-NHL of various subtypes, and explore its correlation to clinicopathologic features and prognosis of T-NHL., Methods: The expression of Mcl-1 protein in 72 specimens of T-NHL was detected by immunohistochemistry. The clinical features, treatments, and outcomes of the T-NHL patients were analyzed retrospectively., Results: The weak positive rates of Mcl-1 were 44.4% in precursor T lymphoblastic lymphoma (T-LBL), 0% in anaplastic large T-cell lymphoma (ALCL), and 18.9% in other peripheral T-cell lymphoma (PTL); the positive rates were 0%, 100%, and 49.1%, respectively (P<0.001). Weak diffuse cytoplasmic staining of Mcl-1 was detected in T-LBL, and strong cytoplasmic staining with perinuclear accentuation was detected in ALCL. The overall survival time was significantly longer in the PTL patients with high Mcl-1 expression than in the PTL patients with weak/negative Mcl-1 expression (>32 months vs. 15 months, P=0.007), and longer in the T-LBL patients without Mcl-1 expression than in the T-LBL patients with weak Mcl-1 expression (21 months vs. 7 months, P=0.58)., Conclusions: The intensities of Mcl-1 expression in T-NHL of various histological subtypes are different. It is specifically highly expressed in ALCL. High expression of Mcl-1 is correlated to better prognosis of PTL.

Background & Objective: Mutation or deletion of PTEN gene is related to a variety of tumors. PTEN gene abnormality is closely related to the tumorigenesis of glioma. This study aimed to investigate the expression of PTEN gene in human glioma cell lines U251 and SHG-44, and explore its effect on cell proliferation., Methods: The expression of PTEN gene in U251 and SHG-44 cells was detected by reverse transcription-polymerase chain reaction (RT-PCR). The recombinant eukaryotic expression vector containing wild-type PTEN gene was transfected into U251 and SHG-44 cells by cation polymex. The stably transfected cells were selected by G418 and amplified. Cell morphology was observed under microscope. The effect of PTEN gene on cell proliferation was assessed. The expression of PTEN protein and glial fibrillary acidic protein (GFAP) were detected by Western blot and immunohistochemistry., Results: Point mutation and deletion of PTEN mRNA were observed respectively in U251 and SHG-44 cells. The proliferation rates of U251 and SHG-44 cells were inhibited by 39.1% and 27.8% of control at 7 days after transfection. The expression of PTEN and GFAP were both increased. The stably transfected U251 cells differentiated toward astrocytes, but SHG-44 cells had no obvious morphologic changes., Conclusion: Restoring expression of wild-type PTEN could induce differentiation of glioma cells differently.

Objective: To study the incidence of PTEN, Mdm2 and p53 expression in different histopathological grades of astrocytoma and the signal transduction mechanism through which PTEN affects Mdm2 and p53 expression., Methods: Surgical specimens from 68 brain astrocytoma patients were analyzed to detect PTEN, Mdm2 and p53 expression by immunohistochemical method., Results: The incidence of PTEN, Mdm2 and p53 expression was 54.4% (37/68), 41.2% (28/68) and 45.6% (31/68). The positive incidence of Mdm2 was 24.3% (9/37) in 37 cases with PTEN expression and 61.3% (19/31) in 31 cases without, which had a negative correlation between Mdm2 expression and PTEN expression (P < 0.01). Eighteen cases showed Mdm2(+)/p53(+) and 27 cases showed Mdm2(-)/p53(-). A significant correlation was found between Mdm2 and p53 expression (P < 0.05)., Conclusion: The histopathological grade of astrocytoma is correlated with the loss of PTEN expression, Mdm2 amplification and p53 mutation. Mdm2 amplification, being in accordance with p53 mutation, can be inhibited by PTEN expression.

Background & Objective: Acute lymphocytic leukemia(ALL), which is sensitive to tumor necrosis factor (TNF) is one of hematological malignances derived from lymphoid tissue. Genetic polymorphisms in the tumor necrosis factor (TNF) locus can affect transcription and expression of TNF genes. This study was designed to investigate the relationship between -308 bp polymorphism in tumor necrosis factor-alpha(TNFalpha) gene and +252 bp in lymphotoxin-alpha(LTalpha) gene and the pathogenesis, clinical course, and outcome of ALL., Methods: The single base mutation polymorphism in TNFalpha gene and LTalpha gene were analyzed among 29 Chinese patients with ALL and 72 normal controls using polymerase chain reaction (PCR)-restrictive fragment length polymorphism (RFLP). The clinical data were collected and survival analysis was performed., Results: The difference of distribution of genotypes, alleles of TNFalpha(-308), LTalpha(+252), and TNF/LT polymorphic extended haplotypes between the ALL patients and control group were not statistically significant (P >0.05). In patients, no statistically significant association was found between the presence of a given TNF/LT haplotype status and clinical characters such as sex, white blood cell (WBC) counts, central nervous system involvement, and the response to therapy(P >0.05). The estimated 1-year overall survival rates in the groups of patients carrying high-risk and low-risk haplotypes were not statistically significant (P >0.05) using Kaplan-Meier method. In multivariate Cox regression models the TNF/LT haplotype status was not found to be a risk factor for outcome (P >0.05)., Conclusion: These data suggest that genetic polymorphisms in the TNFalpha(-308) and LTalpha(+252) are not crucial in the pathogenesis, clinical course, and outcome of ALL patients.