OBJECTIVETo study the effect of treatment of renal anemia by combination of Bushen Jianpi Recipe (BJR, a Chinese experience recipe for supplementing Shen and supporting Pi) and low dosage erythropoietin (EPO), and the influence of treatment on change of serum tumor necrosis factor alpha (TNF-alpha) so as to explore the possible mechanism of integrative Chinese and western medicine (ICWM) in treating renal anemia.

METHODSPatients with renal anemia were randomly divided into two groups, the ICWM group and the control group, symptomatic and supporting treatment such as dialysis, supplementing of ferrous, foliac acid and vitamin B12, was given to both groups. Additionally, to the ICWM group, 50 IU/kg of EPO subcutaneous injection for twice every week, and oral intake of BJR, one dose per day taken in two parts, were given, and to the control group, EPO alone, 50 IU/kg by subcutaneous injecting, 3 times per week, was given. The therapeutic course for two groups was 3 months. Blood levels of hemoglobin (Hb), hematocrit (Hct), TNF-alpha were measured before treatment and the therapeutic effect was observed.

RESULTSAfter treatment, the levels of Hb and Hct increased significantly in both groups (P < 0.01), comparison between the two groups in Hb and Hct after being treated for 3 months showed significant difference (P < 0.05). The serum level of TNF-alpha was markedly higher than normal range in both groups before treatment, it significantly lowered after treatment in the ICWM group (P < 0.05), but unchanged in the control group.

CONCLUSIONCombination of BJR and EPO could significantly inhibit the production of TNF-alpha, this may be an important factor for ICWM in effectively improving sensitivity to EPO.

Adult ; Anemia ; blood ; drug therapy ; etiology ; Chronic Disease ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Erythropoiesis ; drug effects ; Erythropoietin ; therapeutic use ; Female ; Glomerulonephritis ; blood ; complications ; drug therapy ; Humans ; Male ; Middle Aged ; Phytotherapy ; Recombinant Proteins ; Tumor Necrosis Factor-alpha ; metabolism

Objective To explore the prenatal genetic diagnosis for classic phenylketonuria (PKU) families.Methods Probands and their family members from three classic PKU families were analyzed by combining linkage analysis through short tandem repeats (STR) polymorphism and PCR-sequencing for the exons within mutation hot spot of phenylalanine hydroxylase gene.Results Linkage analysis found uninformative for Family 1,while 100 % confirmative information was obtained from Family 2 and 3.Sequencing showed compound heterozygous mutations of phenylalanine hydroxylase gene for all of the three probands.Five mutations were detected,namely Y166X,R243Q,R413P,EX6-96A ＞ G and IVS11-1G＞ C,and IVS11-1G ＞ C was a novel identified muntation.Information from linkage analysis and mutation screening showed clearly that the fetus of Family 1 and 2 were affected,while normal for Family 3.Conclusions For those PKU families,reliable service of prenatal genetic diagnosis could be provided by combining linkage analysis with mutation screening of phenylalanine hydroxylase gene.

Objective To illustrate if the racial coefficient (Rc) be biased by different reference GFR (rGFR) distribution among studies. Methods 1405 white and 321 African American participants in MDRD study and 684 chronic kidney disease(CKD)patients in Chinese eGFR Investigation Study were included.Firstly.the unweighted datasets of white and Chinese were stacked together.rGFR,age and plasma creatinine (Pcr) were log transformed.Linear regression model was constructed using log transformed rGFR as dependent,gender,race and log transformed Pcr and age as independent.Unweighted RC (uRC) for Chinese was calculated.Then.the Chinese CKD distribution of rGFR was weighted to be the same as that in White American.and weighted RC (wRC)for Chinese was calculated.The cases of White.and African-American were stacked together.The cases of African-American were weighted to make the rGFR distribution the same as that in White-American and Chinese population respectively,and RCs for African-American were calculated. Resuits The uRC for Chinese was 1.197(1.180-1.211)and the wRC was 1.130 (1.117-1.143).The two RCs did not overlap with each other.The RCs for African-American were 1.205(1.19-1.219)and 1.233(1.219-1.247)respectively. Conclusions The RCs were influenced by the difference of rGFR distribution.To find out the real RC.an intemational collaborative study iS needed,with the same rGFR measure method.strict control of Pcr measurement,and the same rGFR distribution.

OBJECTIVESTo investigate the cure effect of tumor antigen specific CTL on a model of human hepatocellular carcinoma in nude mice LCI-D20.

METHODSDendritic cells (DCs) were induced from peripheral blood mononuclear cells of healthy people in vitro by using recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) and interleukin-4 (rhIL-4) and were pulsed with tumor antigen from hepatocellular carcinoma cell line MHCC97H. Then tumor antigen specific cytotoxic T lymphocytes (CTLs) were induced. By intraperitoneal injection of tumour antigen specific CTLs into the LCI-D20, the preventive and therapeutic effects of these CTLs to HCC in the LCI-D20 model were assessed. Cytokine-induced killer (CIK) cells and phosphate buffer solution were used as controls at the same time.

RESULTSThe weights of tumors in the tumor antigen specific CTL group, in the CIK cell group and in the blank group were (1.11+/-0.63), (1.12+/-0.36) and (2.68+/-0.53) grams respectively (t = 5.18, t = 6.06, P < 0.01). The amount of blood alpha fetal protein in the tumor antigen specific CTL and CIK groups were (52.1+/-9.7) microg/L and (48.6+/-5.2) microg/L, and was (82.2+/-7.2) microg/L in the blank group (t = 17.26, t = 22.07, P < 0.01 respectively). The metastasis rates in livers were 16.7%, 16.7% and 58.3% in the tumor antigen specific CTL, CIK cell and blank control groups respectively (chi2= 4.44, P < 0.01). The survival time of the mice in the tumor antigen specific CTL group was (79.0+/-5.02) days, (73.3+/-7.0) days in the CIK group, and (52.3+/-5.2) days in the blank group (t = 14.56, t = 17.54, P < 0.01).

CONCLUSIONTumor antigen specific CTLs may prevent metastasis in the LCI-D20 model and prolong the survival time.

Animals ; Antigens, Neoplasm ; immunology ; Carcinoma, Hepatocellular ; immunology ; pathology ; Cell Line, Tumor ; Dendritic Cells ; cytology ; immunology ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Interleukin-4 ; pharmacology ; Liver Neoplasms ; immunology ; pathology ; Male ; Mice ; Mice, Nude ; Neoplasm Metastasis ; Neoplasm Transplantation ; Recombinant Proteins ; T-Lymphocytes, Cytotoxic ; immunology

A rapid and accurate method of DNA typing for HLA was established to compensate the unsatisfactory serological typing for HLA before transplantation. DNA typing for HLA using by reverse polymerase chain reaction with sequence-spcific oligo probe (reverse PCR-SSOP) could detect HLA-A(*0101 - 8001) and B(*07021 - 8201). The results showed that HLA-AB alleles were successfully analysed in 60 matching subjects and 16 control DNAs from UCLA by reverse PCR-SSOP without false negtive and false positive results. The results were concordance with those of UCLA. The error rate of serological typing was 6.4% for HLA-A and 7.4% for HLA-B. The serological typing missed HLA-A24 and HLA-B46 for two patients with leukemia respectively. Our results suggest that DNA typing for HLA by reverse PCR-SSOP has proved to be a high-resolution, high-specificity, rapid and accurate technique, suitable for clinical application with a greater precision than serological typing.

OBJECTIVETo investigate the role of mitogen-activated protein kinases (MAPKs)-extracellular signal regulated kinase1/2 (ERK1/2) signal pathway in the regulation of Compound Danshen Injection (CDI) induced AQP3 expression in the human amniotic epithelial cells (hAECs).

METHODShAECs of term pregnancy with normal amniotic fluid volume (AFV) or isolated oligohydramnios were primarily cultured. And the cells were equally divided into four groups, i.e., the vehicle control group, the U0126 group, the CDI group, the CDI + U0126 group. The expressions of phosphorylated ERK1/2 (p-ERK1/2) and AQP3 in hAECs were detected using Western blot analysis.

RESULTS(1) When compared with the control group, the expression level of p-ERK1/2 in hAECs in those with normal AFV and oligohydramnios obviously decreased in the U0126 group (P < 0.05). The expression level of p-ERK1/2 could be elevated in the CDI group (P < 0.05). The expression level of p-ERK1/2 in hAECs was higher in the CDI +U0126 group than in the U0126 group, but lower in the CDI + U0126 group than in the CDI group (P < 0.05). (2) There was no obvious change in AQP3 expression in hAECs with normal AFV between the U0126 group and the vehicle control group (P > 0.05). There was no statistical difference in the expression level of AQP3 between the CDI group and the U0126 +CDI group (P > 0.05), but they were higher than those in the vehicle control group (P < 0.05). (3) Compared with the vehicle control group, the expression level of AQP3 in hAECs with oligohydramnios significantly decreased in the U0126 group and increased in the CDI group (P < 0.05). The expression level of AQP3 was lower in the U0126 + CDI group than in the CDI group, but higher in the U0126 +CDI group than in the U0126 group (P < 0.05).

CONCLUSIONCDI could regulate AQP3 expression level in hAECs with oligohydramnios via activating the MAPK-ERK1/2 signal transduction pathway.

Amnion ; cytology ; Aquaporin 3 ; metabolism ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; drug effects ; metabolism ; Female ; Humans ; MAP Kinase Signaling System ; Phenanthrolines ; pharmacology

Objective To investigate the effect of collaborative assessment and management on skin of high-risk pressure ulcer patients by nurses from operating room and wards. Methods 709 high-risk pressure ulcer patients who underwent surgical operations in our hospital were selected as the control group from January to June 2016.The 3S pressure ulcer risk assessment form was used to evaluate the skin of the operated patients.Another 680 high-risk pressure ulcer patients were selected as the observation group from July to December in 2016.The skin evaluation and pressure ulcer protection measures in the observation group were the same as those in the control group.The assessment was done by the nurses from the operating room and the wards separately.According to the evaluation form, the nurses in the operating room explained the patient's skin condition to the ward nurses in details, and then they cooperated in managing the postoperative skin of the patients. The two groups were compared in respect of the incidence of pressure ulcers at the time points of postoperative 24h,48h and 72h. Results There was no significant difference in the incidence of pressure ulcers after operation between the two groups(P>0.05).The incidence of pressure ulcers in the observation group was significantly lower than that of the control group at the time points of 24h, 48h and 72h after operation (P<0.05). Conclusion The collaborative assessment and management of skin for surgical patients with high-risk pressure ulcer by the nurses from both operating room and wards can effectively prevent the occurrence and development of pressure ulcers,worthy of clinical application.

Insulin-like growth factor binding-protein-7 (IGFBP7) was obtained from our previous colonic adenocarcinoma (CRC) and normal mucosa suppression subtraction hybridization (SSH) cDNA libraries. By RT-PCR and immunohistochemistry, we found that IGFBP7 was overexpressed in CRC tissue compared to normal tissue. However, our in vitro experiments performed in 10 CRC cell lines showed that IGFBP7 expressed only in SW480 and Caco2 cell lines, which implied an underlying reversible regulatory mechanism. Using methylation-specific PCR (MSP) and bisulfite sodium PCR (BSP), we found that its expression was associated with DNA hypomethylation of exon1. This was further supported by the in vitro study which showed restored IGFBP7 expression after demethylation agent 5-aza-2'-deoxycytidine treatment. Correlation analysis between IGFBP7 expression and prognosis indicated that overexpression of IGFBP7 in CRC tissue correlated with favourable survival. Investigation of the functional role of IGFBP7 through transfection studies showed that IGFBP7 protein could inhibit growth rate, decrease colony formation activity, and induce apoptosis in RKO and SW620 cells, suggesting it a potential tumor suppressor protein in colorectal carcinogenesis. In conclusion, our study clearly demonstrated that IGFBP7 plays a potential tumor suppressor role against colorectal carcinogenesis and its expression is associated with DNA hypomethylation of exon 1.
Adenocarcinoma ; genetics ; metabolism ; Apoptosis ; genetics ; Cell Line, Tumor ; Colorectal Neoplasms ; genetics ; metabolism ; DNA Methylation ; Exons ; Gene Expression Regulation, Neoplastic ; genetics ; Humans ; Insulin-Like Growth Factor Binding Proteins ; genetics ; metabolism ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

BACKGROUNDIn the process of hepatic fibrosis, the accumulation of collagen fibers is strongly related to the hepatic function. The aim of this study was to investigate the three-dimensional architecture of the collagen network in the liver of rats with hepatic fibrosis.

METHODSHealthy adult male Wistar rats (n = 32) were randomly divided into a control group (n = 16) and a hepatic fibrosis group (n = 16). In the control group, the rats were treated with peanut oil while the rats in hepatic fibrosis group were treated for 10 weeks with 60% CCl(4) diluted in peanut oil. The quantity of collagen fibers was detected by Western blotting; distribution of the collagen was detected by sirius red staining and polarized microscope; the three-dimensional architecture of collagen in the liver was observed under the scanning electron microscope after fixed tissues were treated with cell-maceration using NaOH. Statistical analysis was performed using the u test.

RESULTSThe quantity of collagen fibers increased significantly in the hepatic fibrosis group. With the aggravation of hepatic fibrosis, collagen fibers gradually accumulated. They interlaced the reticulation compartment and formed a round or ellipse liver tissue conglomeration like a grape framework that was disparate and wrapped up the normal liver lobule. The deposition of collagen fibers was obvious in adjacent hepatic parenchyma, especially around the portal tracts.

CONCLUSIONOur experiment showed the collagen proliferation and displays clearly the three-dimensional architecture of collagen fibers in rat liver with hepatic fibrosis by scanning electron microscope. It can provide a morphological foundation for the mechanisms of changed haemodynamics and portal hypertension in hepatic fibrosis.

Animals ; Blotting, Western ; Collagen ; ultrastructure ; Liver Cirrhosis, Experimental ; pathology ; Male ; Microscopy, Electron, Scanning ; Rats ; Rats, Wistar

Objective To observe the effect of electroacupuncture(EA)at Baihui(GV20)and Siguan(Hegu/LI4 and Taichong/LR3)of affected side on expression of Tax1-binding protein 1(TAX1BP1)in cerebral cortex in focal cerebral ischemia-reperfusion rats,so as to in-vestigate its protective mechanism in inhibiting nuclear factor kappa B(NF-κB)signaling pathway and promoting neurobehavioral recovery. Methods A total of 105 Sprague-Dawley rats were randomly assigned into sham group,model group and EA group.Each group was ran-domly assigned into reperfusion six hours,twelve hours,24 hours,48 hours,72 hours groups after two hours of ischemia.The model was es-tablished by right middle cerebral artery occlusion and reperfusion.EA group received electroacupuncture at Baihui and left Siguan(Hegu and Taichong)acupoints.Neurobehavioral evaluation,TAX1BP1 protein expression,TAX1BP1 positive cell count,zink finger protein A20 expression, and nuclear NF-κB p65 protein expression were tested in each group. Results There was no neurological deficit in the sham group. Compared with the model group, the neurological scores at 48 hours, 72 hours after reperfusion decreased in EA group (P<0.05). Compared with the control group,the TAX1BP1 expression at twelve hours,24 hours and 48 hours after reperfusion increased in the model group(P<0.05),and further increased at twelve hours,24 hours,48 hours and 72 hours after reperfusion in EA group(P<0.05),and peaked at 24 hours after reperfusion.Compared with the sham group,the expression of A20 and NF-κB p65,and the number of TAX1BP1 positive cells increased in the model group(P<0.05),and the expression of A20 and the number of TAX1BP1 positive cells further increased,(P<0.05)and the expression of NF-κB p65 decreased in EA group(P<0.05)at 24 hours after reperfusion.Immunofluorescence labeling indicat-ed that TAX1BP1 protein primarily expressed in the cytoplasm,TAX1BP1 protein and A20 protein co-expressed in the cytoplasm.Immuno-histochemistry showed indicated that NF-κB p65 mainly expressed in the nucleus in the model groupr,and mainly expressed in the cyto-plasm in the EA group.Conclusion Electroacupuncture could significantly inhibit neuronal NF-κB signaling pathway and promote neurobe-havioral recovery in focal cerebral ischemia-reperfusion,which may related with up-regulating TAX1BP1 protein expression.