[The effect of equilibrium psychological intervention in wounded patients in Lushan earthquake].

OBJECTIVE: To investigate the early psychological effect of the equilibrium psychological intervention on the person who were injured in the disaster incident. METHODS: The equilibrium psychological intervention was used to the injured person in the Lushan earthquake during the early period. The GHQ-12, HAMA and HAMD were used before and after the evaluation. RESULTS: The score of the GHQ-12 decreased from (3.488 +/- 2.900) to (1.610 +/- 0.840), which showed the significant differences (P < 0.001). The total score of the HAMA and the score of the somatic anxiety factor and mental anxiety factor decreased significantly, compared with the base line (P < 0.001 respectively). The total score of the HAMD and the score of the sretardation factor, somatization factor and sleep disorder factor also decreased significantly, compared with that of the base line (P < 0.005 respectively). CONCLUSION: The equilibrium psychological intervention has the positive effect on the persons who were injured in the disaster incident during the early period.

Does the reporting of randomized clinical trials published in Chinese pediatrics journals improve after the CONSORT Statement is adopted?

BACKGROUND: There is no systematic assessment whether the quality of reporting has been improved since the CONSORT Statement was introduced into China in 1997. The aim of this study is to determine whether the use of the CONSORT Statement is associated with improved quality of reporting of RCTs published in Chinese pediatrics journals. METHODS: Six core Chinese pediatrics journals that included Journal of Clinical Pediatrics, Chinese Journal of Contemporary Pediatrics, Chinese Journal of Practical Pediatrics, Chinese Journal of Evidence-based Pediatrics, Chinese Journal of Pediatrics, and Chinese Journal of Pediatric Surgery were searched from inception through Dec. 2010. The CONSORT checklists were used to assess the quality of reporting. Data was collected using a standardized form. Analyses were performed using SPSS 15.0 software. RESULTS: A total of 619 RCTs were included. The quality of reporting has improved significantly in aspects such as introduction, recruitment, baseline data, and ancillary analyses (p<0.05), but not in several important methodological components, including sample size calculation (0.63% vs.1.08%), randomization sequence generation (3.18% vs. 7.58%), allocation concealment (0% vs. 1.08%), and blinding (0% vs. 0.87%). CONCLUSIONS: The quality of reporting of RCTs has not significantly improved since the CONSORT Statement was introduced into China. The reporting remains poor, and often inadequate for assessment of the rigor of studies. Chinese pediatrics journals should reinforce the use of the CONSORT Statement in the reporting of trials.

[The killing effect and bystander effect of linamarase/linamarin suicide gene system on human hepatocellular carcinoma cell line HepG2 in vitro].

AIM: To investigate the killing effect of linamarase/linamarin (lis/lin) system on hepatocellular carcinoma cell line HepG2 in vitro. METHODS: A cDNA library was built from RNA of cassava by RT-PCR, then the linamarase gene was amplified from it by PCR and cloned into the eukaryotic expression vector plasmid pEGFP-N1, which made up the recombinant plasmid pEGFP-N1-lis. The human HCC cells HepG2 were transfected with the recombinant plasmid mediated by electroporation and screened by G418 to yield the positive clone which was termed HepG2/lis. The expression of lis was confirmed by fluorescent staining, RT-PCR and Western blot. The killing effect and bystander effect of linamarin with different concentrations on HepG2 was detected by MTT. RESULTS: RT-PCR confirmed the expression of lis gene in HepG2 and Western blot analysis confirmed existence of lis-EGFP fusion protein in HepG2. Linamarin in low concentration had shown notable cytotoxic effect on HepG2/lis. When HepG2/lis cells were mixed with parental HepG2 cells at a ratio of 10:90 and cultivated in 500 mg/L lin medium, significant bystander effect was observed in vitro. CONCLUSION: The linamarase/linamarin suicide gene system has strong killing effect and bystander effect on HCCs with the concentration of 500 mg/L lin.

Decreased estradiol release from astrocytes contributes to the neurodegeneration in a mouse model of Niemann-Pick disease type C.

Niemann-Pick disease type C (NPC) is a deadly neurodegenerative disease often caused by mutation in a gene called NPC1, which results in the accumulation of unesterified cholesterol and glycosphingolipids in the endosomal-lysosomal system. Most studies on the mechanisms of neurodegeneration in NPC have focused on neurons. However, the possibility also exists that NPC1 affects neuronal functions indirectly by acting on other cells that are intimately interacting with neurons. In this study, using a heterotypic neuron-glia coculture system, we found that wild-type neurons cultured on a layer of NPC1-/- astrocytes showed decreased neurite growth compared with those cultured on wild-type astrocytes. RT-PCR and immunohistochemical assessments showed significantly lower expression of neurosteroid enzymes and StAR (steroidogenic acute regulatory protein) in NPC1-/- astrocyte cultures than in wild-type cultures. Furthermore, a reduced level of estradiol was measured from both astrocyte culture medium and whole brains from NPC1-/- mice. Administration of 17beta-estradiol to neonatal NPC1-/- mice significantly delayed the onset of neurological symptoms, increased Purkinje cell survival, and extended the animals' life span. Our findings suggest that astrocyte dysfunction contributes to the neurodegeneration of NPC and estradiol treatment may be useful in ameliorating progression of the disease.

Autocrine expression of hepatocyte growth factor and its cytoprotective effect on hepatocyte poisoning.

AIM: To construct pEGFP-hepatocyte growth factor (HGF) expression vector, the to detect its expression in transfected human hepatocytes, and to investigate the influence of autocrine HGF expression on the proliferative potential and cytoprotective effects in human hepatocytes. METHODS: Human HGF cDNA was ligated to the pEGFP vector. Recombinant plasmid was transfected into human hepatocyte line QZG with liposome. Expression of HGF protein was observed by fluorescence microscopy and immunohistochemistry. Hepatic cells were collected 24, 48, and 72 h after transfection to detect the number of ((3)H)-TdR uptake in DNA. DNA synthesis was observed by using PCNA stain immunohistochemistry. Acute liver cell damage was induced by carbon tetrachloride. Cytoprotective effect was observed by examining the survival rate of hepatocytes and leakage of intracellular alanine transaminase (ALT) and potassium ions. RESULTS: HGF identification of pEGFP-HGF by enzyme digestion showed that HGF fragment was cloned into BamH I and Sal I sites of pEGFP-N3. Expression of GFP in transfected hepatocytes was observed with fluorescence microscopy. The ((3)H)-TdR uptake became 7 times as many as in the control group 96 h after transfection. After HGF transfection, the survival rate of hepatocytes poisoned by CCl(4) significantly increased (83% vs 61%, P<0.05), and the leakage of intracellular alanine transaminase and potassium ions decreased (586 nkat/L vs 1 089 nkat/L, P<0.01; and 5.59 mmol/L vs 6.02 mmol/L, P<0.01 respectively). Culture of transfected hepatic cells promoted the proliferation of other non-transfected cells. CONCLUSION: Transfected HGF is expressed in hepatic cells and has the activity of promoting cell division and protecting hepatic cells against poisoning.

[Migration and differentiation of exogenous rat mesenchymal stem cells engrafted into normal and injured hearts of rats].

OBJECTIVE: To investigate the effects of different microcircumstances on the migration and differentiation of grafted rat mesenchymal stem cells (rMSC) in host myocardium and the feasibility of treatment of myocardial infarction by exogenous adult stem cells. METHODS: rMSC were isolated from the femurs and tibiae of a male Wistar rat and then purified, made into cell suspension, and labeled with DAPI. 35 female Wistar rat were divided randomly into four groups: acute myocardial infarction control group (AMI group, n = 10, the descending anterior branch of left coronary artery was ligated), acute myocardial infarction + rMSC transplantation group (AMI + rMSC group, n = 10, 1 - 3 hours after the ligation DAPI-labeled rMSC were injected into the peri-infarct tissues), normal heart + rMSC transplantation group (normal heart + MSC group, n = 10, DAPI-labeled rMSC were injected into the corresponding myocardium), and mono-nuclear cells transplantation group (AMI + MNCS, n = 5 DAPI-labeled mononuclear cells were injected into he periinfarct tissues). Ten weeks after the implantation, the rats were killed and their hearts were harvested. Immunohistochemistry was used to examine the troponin, GATA-4 and connexin-43. RESULTS: No lymphocyte proliferation and immonologic rejection were seen in the cardiac tissues of the rats implanted with rMSC. DAPI-labeled rMSC with blue nuclei were distributed extensively in the myocardium of the AMI + rMSC group, ovoid in shape and arranged in parallel with the cardiac muscle fibers, and were distributed sporadically like islands in the myocardium of the normal heart + rMSC group, irregular in shape and not arranged in parallel with the cardiac muscle fibers. No blue nucleus was seen in the cardiac tissues of the hearts implanted with DAPI-labeled mononuclear cells. Troponin and GATA4 were positive immunohistochemically in the implanted rMSC with blue nuclei and the host cardiac muscle cells of the AMI group and AMI + rMSC group, however, were negative in the implanted rMSC with blue nuclei and normal cardiac muscle cells of the normal heart + rMSC group. CONCLUSION: Purified rMSC are immunologically tolerable and can be used as donor cells for exogenous cells therapy. Capable of surviving and homing in both in normal and injured hearts, exogenous rMSC migrate and differentiate into cardiac muscle cell-like cells in myocardium with infarction, however, not in normal heart.

Palliative surgery for hilar cholangiocarcinoma.

OBJECTIVE: To evaluate the results of palliative surgical treatment of hilar cholangiocarcinoma in terms of quality of life, survival period and cholangitis rate. METHODS: The clinical data on 232 patients with hilar cholangiocarcinoma in the last 22 years were analyzed retrospectively. Palliative operations included extrahepatic or intrahepatic choledochojejunostomy (123 patients), bridge internal drainage (15), endoscopic biliary drainage (49), percutaneous transhepatic biliary drainage or celiotomy biliary drainage (29), and exploratory celiotomy external drainage (16). RESULTS: In this series, the operative mortality rate was 9.1%, and no significant difference was observed between groups. The rate of cholangitis after operation was significantly lower in Roux-en-Y choledochojejunostomy group (16.2%) and bridge internal drainage group (15.4%) than in internal drainage group (35.5%, P<0.01), including percutaneous transhepatic biliary drainage (PTBD), endoscopic retrograde biliary drainage (ERBD), and celiotomy (or PTBD) external biliary drainage group (39.1%, P<0.01). No significant difference in survival was observed between the Roux-en-Y choledochojejunostomy group (9.3+/-1.8 months) and PTBD (or ERBD) internal drainage group (8.7+/- 2.2 months), but the survivals of the above groups were significantly longer than those of the bridge internal drainage group (6.5+/-1.7 months, P<0.05) and celiotomy (or PTBD) external biliary drainage group (4.4+/-2.1 months, P<0.01). CONCLUSIONS: In unresectable cholangiocarcinomas, either operative bilioenteric bypass or percutaneous transhepatic biliary drainage can achieve significant palliation. Roux-en-Y choledochojejunostomy is the best choice for palliative operation. The use of U-tube is recommended for internal radiation therapy.

[Development of the human/rat chimera model with neonatal rats].

The purpose of this study was to transplant neonatal rat with human cord blood Lin(-) cells to test the possibility of this xenograft model. The Lin(-) cells were purified from human cord blood (CB) using negative selection strategy based on different lineage-specific antigens. The Lin(-) cells were injected into the liver of neonatal rats using a microinjector at an average of 5 x 10(5) cells for each. Peripheral blood (PB) and spleen were collected at 2,4 and 8 weeks after injection. Flow cytometry was performed to detect human cells in the rat PB, PCR was used to detect human cells in PB as well as spleen. The results showed that a definite proportion of human cells existed in peripheral blood of chimeric rat and the human specific beta2 microglobulin gene fragments were detected in spleen genomic DNA of chimeric rat. It is concluded that human/rat chimera model can be developed with neonatal rats. Human/rat xenograft model may provide a useful and convenient method for human hematopoietic stem cell assay in vivo.

[Migration and differentiation of human bone marrow mesenchymal stem cells in the rat brain].

Bone marrow mesenchymal stem cells (MSCs) are multipotent tissue stem cells that can be induced in vitro to differentiate into a variety of cells such as osteoblasts, chondrocytes and adipocytes. MSCs are useful vehicles for both cell and gene therapy for a variety of diseases. Here, we injected human MSCs with enhanced green fluorescent protein (EGFP) into the striatum of Parkinson disease (PD) rat and examined their survival, migration, differentiation, and the behavior changes in PD rats, which will provide a theoretical foundation and technical method for clinic PD therapy by stem cells. The results showed that human bone marrow MSCs can survive in rat brain for a long time (exceeding 70 d). MSCs were found in multiple areas of the rat brain including the striatum, the corpus callosum, contralateral cortex and even the brain vascular wall. Immunocytochemical staining suggested that implanted cells expressed human neurofilament (NF), neuron-specific enolase (NSE) and glial fibrillary acid protein (GFAP). At the same time, remission in abnormal behavior of the PD rats appeared. Rotation scores decreased gradually from 8.86+/-2.09 r/min pre-transplantation to 4.87+/-2.06 r/min 90 d post-transplantation (statistic result showed P<0.05).

[Effect of granulocyte colony-stimulating factor on the ratio of peripheral blood dendritic cell subsets].

AIM: To clarify the effect of granulocyte colony-stimulating factor (G-CSF) on the radio of two peripheral blood dendritic cell(DC) subsets in-vivo. METHODS: Various doses (0, 5, 10 and 15 microg) of G-CSF were subcutaneosly injected respectively into BALB/c mice, once each day, for 6 days running. Six days later, DCs were separated from peripheral blood. Then the flow cytometry was used to detect the radio of DC1 and DC2 (CD11c(+) CD8a(-) and CD11c(+) CD8a(+)), and to calculate their absolute numbers. RESULTS: After stimulation with varying doses of G-CSF for 6 days, the absolute number of DC2s increased from 9.6x10(6)/L to 55.1x10(6)/L (P<0.01), while that of DC1s had no notably variation, so the ratio of DC1 and DC2 decreases from 4.2+/-1.1 to 0.7+/-0.3 (P<0.01). CONCLUSION: G-CSF can increase the absolute number of peripheral blood DC2s, but has no influence on peripheral blood DC1 number, thus inversing the ratio of DC1/DC2 in-vivo.