OBJECTIVETo study the effects of Kangai injection on the enzyme activities of macrophages and morphology of spleen and thymus from rats.

METHODSTwenty four male SD rats were randomly divided into two groups (n = 12), normal control group and experimental group. The rats in experimental group were injected with Kangai injection at the dosage of 5 ml/kg x d for 30 days peritoneally and those in control group were injected with nomal saline at the same volume. The content of supermicro protein was assayed by BCA method, the activities of lactate dehydrogenase (LDH), glutathione peroxidase(GSH-Px), and inducible nitric oxide synthase (iNOS) from alveolar macrophages(AM) and peritoneal macrophages (PM) were detected biochemically. The activities of acid phosphatase (ACP), superoxide dismutase(SOD) and succinate dehydrogenase (SDH) from AM and PM were detected by ELISA. The morphology of spleen and thymus were observed by light microscopy.

RESULTSThe activities of LDH, GSH-Px and iNOS within AM and PM from experimental group were increased significantly compared with those of control group (P < 0.05). The activities of ACP, SOD and SDH in AM and PM from experimental group were also higher than those from control group (P < 0.05). Microscopically, there was thickening of peripheral arterial lymphatic sheath, enlargement of splenic lymphoid nodules with expended germinal center in the spleen of experimental group. There was no significant difference in the mophology of thymus between the two groups.

CONCLUSIONKangai injection may improve immune function by activating macrophages.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Glutathione Peroxidase ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Macrophages ; drug effects ; enzymology ; Male ; Nitric Oxide Synthase Type II ; metabolism ; Rats ; Rats, Sprague-Dawley ; Succinate Dehydrogenase ; metabolism ; Superoxide Dismutase ; metabolism

AIM: To observe thickness and morphological changes of bulbar conjunctiva pre- and post epidemic hemorrhagic conjunctivitis ( EHC ) therapy by optical coherence tomography ( OCT) .
METHODS: Observed morphological changes and measured the bulbar conjunctiva thicknesses of 29 cases (36 eyes) of incipient (1-2d) EHC patients, who were received and treated by department of ophthalmology, the Putuo Affiliated Hospital of Shanghai University of Traditional Chinese Medicine from May 2013 to December 2013, by OCT. Then measured the thickness again on 7, 14d after the therapy.
RESULTS: Among 29 patients (36 eyes), 7d after the EHC therapy, in 27 cured eyes, the full-thickness ( before 344. 00±59. 91μm, after 230. 19±22. 16μm, t=11. 75, P<0-01); epithelial thickness ( before 56. 52±6. 19μm, after 51. 37±5. 53μm, t=4. 61, P<0. 01); and stromal thickness (before 287. 11±60. 56μm, after 178. 81±20. 20μm, t=10. 69, P<0. 01) of patients' bulbar conjunctiva were thicker than values measured after therapy with significant difference. Significant difference was also found for full-thickness ( before 361. 39±65. 56μm, after 233. 44±22. 57μm, the difference was statistically significant, t=14. 45, P<0. 01);epithelial thickness ( before 55. 50±6. 72μm, after 46. 67±5-24μm, t=10. 06, P<0. 01) and stromal thickness ( before 305. 61±66. 02μm, after 186. 78±21. 82μm, t=13. 11, P<0-01 ) of patients' bulbar conjunctiva between values measured before and 14d after therapy.
CONCLUSION: The OCT is able to measure the thickness of bulbar conjunctiva in EHC patients. An significant increase was found in full, epithelial and stromal thickness of EHC patients' bulbar conjunctiva. With recovery from the disease, subepithelial fluid, interlaminar fluid and edema of the bulbar conjunctival stroma faded away firstly, which provide references for clinical therapies of the EHC.

Transgenic tobacco plants expressing a stress responsive gene BoRS1, isolated from Brassica oleracea var. acephala, under the control of the 35S promoter of the Cauliflower mosaic virus were produced. Some plants were further used to test the effect of high level BoRS1 expression on drought stress resistance. The presence of transgene in putative transgenic plants was confirmed by PCR analysis. Thirty-six among 130 transformants showed amplification of predicted fragment of BoRS1 while no amplification was observed in the control. Some transgenic lines confirmed by PCR analysis were analyzed through semi-quantitative one-step RT-PCR for the expression of BoRS1 gene. Amplification of 1.4 kb cDNA product revealed transcription of BoRS1 gene. Meanwhile, differential intensity of the cDNA band indicated variable expression levels of the transgene among different transformed lines. The water loss of detached leaves from the transgenic plants was slower than that of the control. Transgenic tobaccos and the non-transgenic controls were used for further drought stress experiments by using different concentration of mannitol. The transformants showed higher tolerance to drought stress than non-transgenic plants and different transgenic lines exhibited different tolerance during drought stress. These results showed that the BoRS1 gene probably play role in enhancing the ability to drought stress.
Adaptation, Physiological ; genetics ; Agrobacterium tumefaciens ; genetics ; Brassica ; genetics ; Dehydration ; Genes, Plant ; Plant Proteins ; genetics ; metabolism ; Plants, Genetically Modified ; genetics ; growth & development ; physiology ; Stress, Physiological ; Tobacco ; genetics ; growth & development ; physiology ; Transformation, Genetic ; Transgenes

Objective To investigate the protective effect of complement 5a receptor 1 (C5aR1) antagonist on ascending urinary tract infection in mice.Methods (1) Female C57BL/6 mice were randomly divided into experimental and control groups:38 mice in each group,and inoculated with E.coli by urethral catheterization to set up the ascending urinary tract infection model.C5aR1 antagonist (W54011 or PMX53) and corresponding control (PBS or control peptide) were initially given either at 2 h before or 3 h after infection by intraperitoneal injection.Mice were sacrificed to assess the infection in bladder and kidney at 24 or 48 h after infection.The bacterial load of bladder and kidney tissue was measured by agar plate assay.The mRNA expression of renal inflammatory factors was detected by real-time RCR.The renal tissue injury and inflammatory cell infiltration were assessed by HE staining and pathological scores.(2) Primary cultured renal tubular epithelial cells were randomly divided into antagonist and control groups to detect and compare the bacterial adhesion to renal tubular epithelial cells in vitro.Results Compared with control groups,the initial delivery of C5aR1 antagonist (W54011 or PMX53) before E.coli inoculation reduced the bacterial load in bladder and kidney tissue 48 h after infection (all P ＜ 0.01).In experimental group given W54011 before infection,the renal pathological scores were reduced (both P ＜ 0.05),as well as renal inflammatory factor expressions:CXCL-1 mRNA,IL-6 mRNA and TNF-o mRNA (all P ＜ 0.05).Compared with corresponding control groups,the initial delivery of PMX53 after E.coli inoculation could also reduce the bacterial load in bladder and kidney tissue 48 h after infection (both P ＜ 0.01).Furthermore,C5aR1 antagonists W54011 and PMX53 could decrease bacteria adhesion to renal tubular epithelial cells in vitro,compared with control groups (both P ＜ 0.05).Conclusions C5aR1 antagonists can significantly attenuate renal tissue injury,ameliorate renal inflammation and the adhesion of bacteria to renal epithelial cells.C5aR1 may be an effective target for the prevention and treatment of urinary tract infection.

PURPOSE: Although the interferon α (IFNα) signaling and the paired-like homeodomain transcription factor 2 (PITX2) have both been implicated in the progression of breast cancer (BCa), it remains obscure whether these two pathways act in a coordinated manner. We therefore aimed to elucidate the expression and function of PITX2 during the pathogenesis of endocrine resistance in BCa. MATERIALS AND METHODS: PITX2 expression was assessed in BCa tissues using quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunohistochemistry and in experimentally induced letrozole-resistant BCa cells using RT-qPCR and immunoblotting. Effects of PITX2 deregulation on BCa progression was determined by assessing MTT, apoptosis and xenograft model. Finally, using multiple assays, the transcriptional regulation of interferon-inducible transmembrane protein 1 (IFITM1) by PITX2 was studied at both molecular and functional levels. RESULTS: PITX2 expression was induced in letrozole-resistant BCa tissues and cells, and PITX2 induction by IFNα signaling powerfully protected BCa cells against letrozole insult and potentiated letrozole-resistance. Mechanistically, PITX2 enhanced IFNα-induced AKT activation by transactivating the transcription of IFITM1, thus rendering BCa cells unresponsive to letrozoleelicited cell death. Additionally, ablation of IFITM1 expression using siRNA substantially abolished IFNα-elicited AKT phosphorylation, even in the presence of PITX2 overexpression, thus sensitizing BCa cells to letrozole treatment. CONCLUSION: These results demonstrate that constitutive upregulation of PITX2/IFITM1 cascade is an intrinsic adaptive mechanism during the pathogenesis of letrozole-resistance, and modulation of PITX2/IFITM1 level using different genetic and pharmacological means would thus have a novel therapeutic potential against letrozole resistance in BCa.
Apoptosis ; Breast Neoplasms ; Breast ; Cell Death ; Heterografts ; Immunoblotting ; Immunohistochemistry ; Interferons ; Phosphorylation ; Polymerase Chain Reaction ; Reverse Transcription ; RNA, Small Interfering ; Transcription Factors ; Transcriptional Activation ; Up-Regulation

OBJECTIVE: To study the expression and effect of Pim1 in primary cortical neurons after hypoxic-ischemic injury. METHODS: Cortical neurons were isolated from 1-day-old C57BL/6 mice and cultured in neurobasal medium. On the 8th day of neuron culture, cells were subjected to oxygen-glucose deprivation/reoxygen (OGD/R) treatment to mimic in vivo hypoxic injury of neurons. Briefly, medium were changed to DMEM medium, and cells were cultured in 1% O for 3 hours and then changed back to normal medium and conditions. Cells were collected at 0 hour, 6 hours, 12 hours and 24 hours after OGD/R. Primary neurons were transfected with Pim1 overexpression plasmid or mock plasmid, and then were exposed to normal conditions or OGD/R treatment. They were named as Pim1 group, control group, OGD/R group and OGD/R+Pim1 group respectively. Real-time PCR was used to detect Pim1 mRNA expression. Western blot was used to detect the protein expression of Pim1 and apoptotic related protein cleaved caspase 3 (CC3). TUNEL staining was used to detect cell apoptosis. RESULTS: Real-time PCR and Western blot results showed that Pim1 mRNA and protein were significantly decreased in neurons after OGD/R. They began to decrease at 0 hour after OGD/R, reached to the lowest at 12 hours after OGD/R, and remained at a lower level at 24 hours after OGD/R (P<0.01). Overexpression of Pim1 significantly upregulated the protein level of Pim1. Under OGD/R conditions, the CC3 expression and the apoptosis rate in cells of the Pim1 group were significantly lower than in un-transfected cells (P<0.01). CONCLUSIONS Hypoxic-ischemic injury may decrease Pim1 expression in neurons. Overexpressed Pim1 may inhibit apoptosis induced by OGD/R.
Animals ; Glucose ; Mice ; Mice, Inbred C57BL ; Neurons ; Oxygen ; Proto-Oncogene Proteins c-pim-1 ; Rats, Sprague-Dawley

OBJECTIVETo scan for mutations of polycystic kidney disease 1 gene (PKD1) in Chinese population in order to find some features about Chinese patients and a better approach to detect mutations.

METHODSTwenty-five PKD-affected individuals from twenty-one unrelated genealogies and sixteen controls participated in the study. Thirty-five blood samples and six tissues were obtained after receiving informed consent and were in accordance with institutional ethical guidelines. Genomic DNA was isolated from peripheral blood using standard procedures. PCR amplification of genomic DNA was performed to generate the aimed fragments. Amplified fragments were analyzed by denaturing gradient gel electrophoresis (DGGE). A GC clamp was attached to the 5' primer. After that, the abnormal fragments were sequenced on freshly amplified specific PCR products with the dideoxynucleotide chain termination method. Sequencing was performed for all samples to evaluate DGGE.

RESULTSAimed fragments of exons 44 and 45 were amplified. DGGE detected eleven abnormal PCR fragments. Two novel mutations were identified by sequencing, included one nonsense mutation (C12217T) and one frameshift (12431delCT). In addition, one polymorphism (A50747C) was identified. The mutation detection rate is 8% in our study.

CONCLUSIONTwo novel pathogenic mutations were detected, including one nonsense mutation (C12217T) and one frameshift (12431delCT).

Asian Continental Ancestry Group ; genetics ; Codon, Nonsense ; DNA Mutational Analysis ; Exons ; genetics ; Family Health ; Female ; Frameshift Mutation ; Genotype ; Humans ; Male ; Middle Aged ; Mutation ; Pedigree ; Phenotype ; Polycystic Kidney, Autosomal Dominant ; genetics ; Polymorphism, Single Nucleotide ; Proteins ; genetics ; TRPP Cation Channels

OBJECTIVETo discuss the proper surgical management of pancreatic benign and low-grade malignant potential neoplasm.

METHODSThe experience of 72 cases who accepted organ preserving pancreatectomy from January 1990 to May 2010 was analyzed retrospectively. There were 24 male and 48 female, aged from 15 to 68 years with mean age of 46 years. There were 9 cases underwent duodenum-preserving resection of the head of the pancreas, 29 cases underwent spleen-preserving distal pancreatectomy, 11 cases underwent middle segmental pancreatectomy, 23 cases underwent tumor extirpation of huge pancreatic cancer in pancreatic head and body.

RESULTSPancreatic fistula and biliary fistula in 1 case respectively were cured among who accepted duodenum-preserving resection of the head of the pancreas. Pancreatic fistula was found in 3 cases who accepted spleen-preserving distal pancreatectomy. Pancreaticobiliary anastomotic bleeding in 1 case was cured among who accepted middle segmental pancreatectomy. Pancreatic fistula was found in 5 cases among who accepted tumor extirpation of huge pancreatic cancer in pancreatic head and body, and liver metastasis was found in 3 cases at 6, 12, 16 months after surgery respectively.

CONCLUSIONSOrgan preserving pancreatectomy can obviously reduce operative injury to patients, its therapeutic effect is similar to that of classical operation, it is the first option of benign and low-grade malignant potential neoplasm.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Pancreatectomy ; methods ; Pancreatic Neoplasms ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult

Objective:To study the prognostic impact of prognostic nutritional index (PNI) before radiotherapy in clinical stage Ⅲ esophageal cancer patients.Methods:We retrospectively reviewed 125 esophageal cancer patients with clinical stage Ⅲ undergoing definitive radiotherapy in Fourth Hospital of Hebei Medical University from 2013 to 2017. The PNI and nutritional risk index (NRI) were calculated before radiotherapy. The optimal cutoff value of PNI was determined by time-dependent receiver operating characteristics (ROC) at 49.925.The patients were divided into low PNI group(PNI<49.925) and high PNI group (PNI≥49.925). Based on NRI, the patients were divided into normal NRI group (NRI≥100) and abnormal NRI group (NRI<100). Kaplan-Meier method was used to calculate the overall survival (OS) and progression-free survival (PFS) and to perform univariate analysis. The mutlivariate analysis was performed by Cox regression model.Results:PNI was positively correlated with hemoglobin ( r=0.505, P<0.001) and NRI ( r=0.594, P<0.001). The 1-, 3- and 5-year OS rates in the low PNI group were significantly lower than those of the high PNI group (67.5%, 27.3%, 11.4% vs. 85.4%, 45.8%, 27.4%, respectively, χ2=8.569, P<0.05). Moreover, the 1-, 3- and 5-year PFS rates in the low PNI group were obviously higher than those in the high PNI group (59.7%, 23.2%, 4.9% vs. 79.2%, 35.4%, 24.9%, respectively, χ2=6.715, P<0.05). Univariate analysis showed that GTV, radiotherapy dose, chemotherapy, albumin, NRI and PNI were significantly correlated with OS and PFS (OS: χ2=6.822, 4.326, 4.474, 13.123, 8.846, 8.569, P<0.05: PFS: χ2=7.869, 4.636, 5.874, 10.911, 8.544, 6.715, P<0.05). Multivariate analysis showed that GTV, radiotherapy dose and PNI were independent prognostic factors for OS ( P<0.05). And GTV, radiotherapy dose, chemotherapy and PNI were independent prognostic factors for PFS ( P<0.05). Conclusions:The PNI before radiotherapy is a significant and independent predictor for survival of clinical stage Ⅲ esophageal cancer patients. Based on simple and inexpensive standard laboratory measurements, PNI could be a promising prognostic biomarker for esophageal cancer patients.

Objective To investigate the effect and mechanism of transplantation of neuregulin1(NRG1)gene-modified bone marrow mesenchymal stem cells(BMSCs)on the repair of hemi-transected spinal cord injury(SCI)in rats.Methods Isolated and cultured rat BMSCs,followed by transfection with the NRG1 gene.The levels of NRG1 in BMSCs lysate and culture supernatant was deected by ELISA method,and the proliferation activity of the BMSCs was detected by cell counting method.Forty-three healthy 8-week-old SD rats were randomly divided into control group(n=10),SCI model group(n=10),BMSCs group(n=10),and NRG1-BMSCs group(n=13).After establishing the spinal cord hemisection model,animals received in-situ transplantation of BMSCs or NRG1-BMSCs.On the 1,7,14,21,and 28 days after transplantation,the hind limb motor function was evaluated using BBB score and inclined plate test;on the 7th day after transplantation,the migration and distribution of transplanted cells was monitored using a fluorescence microscope;on the 28th day after transplantation,the pathological changes of rat spinal cord tissues was examined using HE staining and Nissl staining;cell apoptosis using TUNEL staining,and levels of endoplasmic reticulum stress-related proteins[X-box binding protein 1(XBP1),C/EBP homologous protein(CHOP),activating transcription factor 4(ATF4),ATF6,glucose-regulated protein 78(GRP78)]and apoptosis-related proteins[B-cell lymphoma-2(Bcl-2)and Bcl-2-associated protein X(Bax)]in rat spinal cord tissues using Western blotting.Results BMSCs were successfully isolated,cultured,and transfected with the NRG1 gene.ELISA method results showed that the NRG1 contents in the NRG1-BMSCs lysate and culture supernatant were significantly higher than that of BMSCs in a time-dependent manner(P<0.05).The proliferation activity of NRG1-BMSCs was significantly higher than that of BMSCs(P<0.05).On the 21 and 28 days after transplantation,the BBB score and the slope angle of the inclined plate in NRG1-BMSCs group were higher than those in SCI model group or BMSCs group(P<0.05).However,it did not reverse to the level in control group(P<0.05).On the 28th day after transplantation,compared with the SCI model group and BMSCs group,neuronal pyknosis reduced,the Nissl body density increased,the expression levels of XBP1,CHOP,ATF4,ATF6,GRP78,and Bax,and the rate of TUNEL-positive cells significantly reduced in NRG1-BMSCs group(P<0.05),and the expression level of Bcl-2 significantly increased(P<0.05).Conclusion Transplantation of NRG1 gene-modified BMSCs can alleviate SCI and improve the recovery of motor function in rats.The mechanism may be related to promoting the proliferation activity of BMSCs,inhibiting cell apoptosis,and mitigating endoplasmic reticulum stress.