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The biogenic monoamine 5-hydroxytryptamine (5-HT) is an ancient intracellular signaling molecule widely distributed in all animals with nervous systems, and has been implicated in principal behaviors. Tryptophan hydroxylase (TRH) induces a highly specific catalytic reaction that converts L-tryptophan (tryptophan) to 5-hydroxy-L-tryptophan (5-HTP) that is subsequently used as a substrate by aromatic L-amino acid decarboxylase (DDC) to form 5-HT. Five-HT is an ancient intracellular signaling molecule that is widely distributed in the animal kingdom and has been implicated in regulating the behaviors of animals with nervous systems. However, the role of TRH in Lepidoptera is not well understood. In this study, we cloned 1 667 bp cDNAs of Bombyx mori TRH (BmTRH), which contains a 1 632 bp open reading frame (ORF). Homology analysis revealed that BmTRH shared high amino acid identity with Homo sapiens TPH and Drosophila TRH (DmTRH). The high homology (70%) of BmTRH with DmTRH suggested that BmTRH could have a function similar to DmTRH. Gene expression analysis revealed that BmTRH was mainly expressed in head and central nervous (CNS). Moreover, immunohistochemistry and Western blotting analyses showed that BmTRH was detected only in larval nervous tissues. Taken together, our results indicate that BmTRH could likely function in the regulation of neural activities in B. mori. The transcripts of B. mori decarboxylase (BmDDC) and B. mori phenylalanine hydroxylase (BmPAH) whose proteins had TRH activity, were also expressed in the CNS tissues, indicating that unlike in Drosophila, two distinct mechanisms likely regulate 5-HT synthesis in silkworm.
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Animals , Amino Acid Sequence , Bombyx , Cloning, Molecular , DNA, Complementary , Insect Proteins , Phenylalanine Hydroxylase , Tryptophan HydroxylaseABSTRACT
Objective To evaluate the early clinical outcomes and complications of oblique lateral interbody fusion (OLIF) in the treatment of degenerative lumbar diseases.Methods All of 83 patients,29 males and 54 females with ages from 32 to 83 (average 60.8± 13.7 y),underwent OLIF with or without posterior pedicle screw-rod instrumentations from October 2014 to February 2017.The index diagnosis was discogenic back pain in 17 cases,spondylolisthesis in 23,lumbar spinal canal stenosis in 25,and degenerative lumbar spinal kyphoscoliosis in 18 cases.The distribution of operative level was 5 at L1,2,13 at L2,3,38 at L3,4,and 69 at L4,5.The mean number of fusion level for each case was 1.5 segments.The operative duration,blood loss during operation,intra-operative and post-operative complications,the length of post-operative hospital stay were recorded.Clinical outcomes were evaluated using visual analogue scale (VAS) and Oswestry disability index (ODI).All patients were followed up for at least 3 months.Lumbar X-ray and CT scans were taken and the clinical outcomes were re-assessed during follow-up.Results Fifty-one in the 83 patients underwent supplementary posterior pedicle screw-rod instrumentation with OLIF procedures.The operation lasted for 43-295 min,with a mean duration of (153 ± 72) min.Mean operation time for each OLIF segment was 43± 12 min.Blood loss during the operation was 30-800 ml,with a mean of 125±74 ml.Mean blood loss for each OLIF segment was 27±13 min.Average length of stay was 5.6 ± 3.2 d,ranging from 3-15 d.The VAS for back pain and leg pain and ODI scores were decreased apparently for each patient.The total incidence of complications was 22.9％ (19/83),including 6.0％ (5/83) of intra-operative complications (4 cases of cage subsidence,1 case of segmental artery injury) and 16.9％ (14/83) of post-operative ones.The latter consisted of ipsilateral hip flexor weakness in 6,ipsilateral anterolateral thigh pain in 2,ipsilateral lateral thigh numbness in 1,contralateral pain in flexion of hip in 1,ipsilateral sympathetic chain injury in 2,and pain in area of iliac bone donor site in 2.All symptoms were released or disappeared during follow-up.Conclusion OLIF as a novel minimally invasive technique can act as a safe and effective treatment for degenerative lumbar diseases,which can also reduce approach-related complications.
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Objective To investigate the protective effect of lentivirus-mediated silencing information regulation protein 6(SIRT6)overexpression on radioactive lung injury in rats.Methods 72 male 150-200 g Wistar rats were randomly divided into three groups(n=24).The 6 MV-X-ray linear accelerator was used to irradiate the lungs.The rats in each group were injected with normal saline(TNF-α)were measured at 1, 2, 4, and 8 weeks after radiotherapy(Lent-SIRT6), and the levels of tumor necrosis factor-α(TNF-IL-6 and IL-1β were measured by HE staining.The levels of TNF-α, IL-6 and IL-1β in liver tissue were detected by HE staining.The levels of TNF-α, IL-6 and IL-1β in liver tissue were detected by HE staining.Results The alveolar wall and alveolar stroma were normal in the control group.The alveolar wall of the irradiated group was thickened and fibrosis, and a large number of hypertrophic fibrous tissues were found in the alveolar stroma.The alveolar wall of Lent-SIRT6 group was thickened And alveolar stromal fibrosis symptoms were lighter than the irradiation group.Compared with the control group, the levels of serum TNF-α and IL-6, neutrophil count and the levels of TNF-α, IL-6 and IL-1β in the liver tissue were significantly higher than those in the control group (P<0.05).The lentiviruses overexpressing SIRT6 could alleviate the abnormalities caused by radiation (P<0.05).The difference was statistically significant (P<0.05).Conclusion SIRT6 can effectively inhibit the inflammatory reaction, reduce the lung injury symptoms of radiation pneumonia, and have some protective and prevention effect on lung injury.
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Objective@#To investigate the incidence, molecular features and clinical significance of RNA splicing machinery genes mutation in myelodysplastic syndromes (MDS) and related diseases.@*Methods@#Mutational analysis of splicing factor 3B subunit 1 (SF3B1) (K700E) , U2 small nuclear RNA auxiliary factor 1 (U2AF1) (S34, Q157P) and serine/arginine-rich splicing factor 2 (SRSF2) (P95) in 118, de novo MDS and related diseases were separately performed by using polymerase chain reaction (PCR) followed by sequence analysis.@*Results@#Of 118 MDS patients, 76 males and 42 females, the median age was 53.5 (13-84) years old. 19.49% (23/118) had SF3B1 (K700E) mutation. As compared with those with wild type SF3B1, patients with SF3B1 K700E were of older[58 (32-78) years vs 51 (13-84) years, z=-1.981, P=0.048], lower HGB level[63 (40-95) g/L vs 77 (34-144) g/L, z=-3.192, P=0.001], higher platelet counts[121 (22-888) ×109/L vs 59 (6-1 561) ×109/L, z=-3.305, P=0.001], lower bone marrow blast cell counts[0.007 (0-0.122) vs 0.017 (0-0.268) , z=-2.885, P=0.004], higher ring sideroblasts percent [0 (0-64%) vs 0 (0-58%) , z=-4.664, P<0.001]. Of 105 MDS patients, 21.9% had U2AF1 (S34, Q157P) mutations. Of 107 MDS patients, 8 patients (7.48%) had SRSF2 (P95) mutations. Patients with SRSF2 mutations were older at diagnosis, the median age was 63 (50-84) years old, including 4 cases RAEB-1. The ratio of mutation was 14.29% (4/28) , and three patients transformed to AML. SF3B1 K700E and SRSF2 P95H mutations coexisted in 1 patient, and SF3B1 K700E and U2AF1 S34Y mutations were found concomitantly in 2 patients.@*Conclusion@#Only SF3B1 gene mutation was closely related to ring sideroblasts, it was the key to pathogenesis of MDS.
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<p><b>OBJECTIVE</b>To investigate the sensitivity and specificity of eosin-5'-maleimide (EMA)assay for the diagnosis of hereditary spherocytosis (HS), and to verify the stability of reagent and samples.</p><p><b>METHODS</b>EMA flow cytometry test, NaCl-osmotic fragility test and acidified glycerol lysis test were performed using peripheral blood samples from 80 patients with HS and 44 patients with other blood diseases, the sensitivity and specificity of the three methods were compared, and the feasibility of EMA binding test was estimated. The stability of EMA reagent and HS samples stored at different temperatures were tested.</p><p><b>RESULTS</b>Among the 124 tested samples, the sensitivity and specificity of EMA binding test was 0.925 and 0.954, that of NaCl-osmotic fragility test was 0.950 and 0.455, and that of acidified glycerol lysis test was 1.000 and 0.318, respectively. Although the sensitivity of NaCl-osmotic fragility test and acidified glycerol lysis test was a little higher than that of EMA binding test, the specificity of the former two methods was poor, they couldn't clearly distinguish whether spherocytosis is hereditary spherocytosis. The experiment results showed that EMA was sensitive to the temperature and should not be stored in a small aliquots at -80 ℃ over a period of 6 months. The stability of the HS sample was better, 6 days storage at 4 ℃ and 3 days storage at room temperature had no influence on the results.</p><p><b>CONCLUSION</b>EMA binding test by flow cytometry showed good sensitivity and specificity for HS diagnosis. EMA reagent should be stored at-80 ℃ and the HS samples should be tested within 6 days storage at 4 ℃ and 3 days at room temperature.</p>
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Humans , Ankyrins , Blood , Eosine Yellowish-(YS) , Flow Cytometry , Hematologic Tests , Sensitivity and Specificity , Spherocytosis, Hereditary , Blood , DiagnosisABSTRACT
Objective To detect the expressions and clinical significance of Nanog and CD44 protein in lung cancer. Methods The expressions of Nanog and CD44 were detected by immunohistochemistry in 50 cases of lung cancer, 32 cases of benign lesion lung tissue and 18 cases of paraneoplastic normal lung tissue. Then their relationships with clinicopathological factors were analyzed. Results The expression of Nanog in lung cancer was significantly higher than those in benign lesion lung tissue and paraneoplastic normal lung tissue (P 0.05). The expressions of Nanog and CD44 in squamous cell carcinomas were higher than those in adenocarcinomas and small cell lung carcinomas (P 0.05). The positive correlation was also noted between the expressions of Nanog and CD44 in lung cancer (r = 0.564, P < 0.05). Conclusion Nanog and CD44 proteins may participate in the genesis and progression of lung cancer. Nanog protein is a potential diagnostic marker and therapeutic target for lung cancer.
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Objective To study the diagnostic value of Sox2 mRNA transcription level in bronchoscopic biopsy specimens from lung cancer patients. Methods The expression of Sox2 mRNA was detected using RT-PCR from 100 hu-man lung cancer biopsy and 18 non-cancer lung biopsy through bronchoscopy. The expression of Sox2 protein was examined by immunohistochemistry from 50 cases of lung cancer biopsy, 32 cases of benign lung lesions and 18 cases of pericarcino-matous normal lung tissues. Then the relationships between Sox2 mRNA transcription level and lung cancer clinical patho-logical parameters were analyzed to test the diagnostic value of Sox2 transcription level. Results The transcription of Sox2 mRNA and its protein expression level were significantly higher in lung cancer than that in benign pulmonary disease tissues (P＜0.05). The transcription of Sox2 mRNA was not correlated with age, gender, histology, lymph node metastasis, TNM stage and differentiation of lung cancer patients (P>0.05). The Sox2 mRNA yielded an area of 0.748 under the ROC curve with the sensitivity of 85.0%and the specificity of 61.1%, taking the cut-off value of 0.513. Conclusion The Sox2 mRNA might be a useful diagnostic marker for lung cancer.
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Objective To detect the proteins levels of RhoA and CDC42 in bone marrow mononucleated cells (BMMC) of patients with primary acute leukemia,and further determine the role of abnormal interactions between hematopoietic progenitor and bone marrow microenvironment on abnormal behaviors of leukemia cells. Methods BMMC samples were separated from 54 primary acute leukemia patients and 22 normal donors and the cell lysis samples were prepared. RhoA and CDC42 proteins were determined by Western blotting. Independent pair T test was conducted to evaluate whether the differences in RhoA and CDC42 expression were statistically significant between leukemia patients and normal donors. Spearman was applied in analyzing the correlation between expression of RhoA and CDC42 proteins and clinical characters of patients. Results RhoA and CDC42 proteins level of primary acute leukemia patients was significantly higher than that of normal samples. Especially, patients with M2,M3 and M5 subtypes exhibited significant higher RhoA proteins levels and M3 subtype exhibited significant higher CDC42 protein levels. Conclusion RhoA and CDC42 protein levels of primary acute leukemia patients are significantly higher than that of normal donors. This result suggests that RhoA and CDC42 associated efficient migration of leukemia cells could be implicated in abnormal interaction of leukemic cell with bone marrow microenvironment.
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<p><b>BACKGROUND AND OBJECTIVE</b>It was proven that Vitamin C could inhibit the growth of many types of tumors as an antioxidant. The aim of this study is to explore role of Vitamin C in proliferation and apoptosis of lung carcinoma cell line A549 and the underlying mechanism.</p><p><b>METHODS</b>A549 cells were cultured in vitro and incubated with Vitamin C. The cell viability was measured by growth curve and clonogentic assay. Flow cytometry was used to analyze cell cycle and detect apoptosis. The levels of expression of Caspase-3 mRNA and Survivin mRNA were detected by RT-PCR.</p><p><b>RESULTS</b>Vitamin C of 400 microg/mL, 4 mg/mL significantly inhibited the growth of A549 cell lines (P = 0.024, P = 0.015, respectively). Flow cytometry showed that the cells major stagnation stayed in the G0/G1 and S phase and the apoptotic rate increased with time prolonged. Vitamin C signifiantly up-regulated the expression of Caspase-3 mRNA, but had no effect on Survivin mRNA.</p><p><b>CONCLUSION</b>Vitamin C can inhibit the proliferation of A549, block A549 cells in G0/G1 and S phase, and induce apoptosis of A549 cells. Apotosis occurred by up-regulated the expression of Caspase-3.</p>
Subject(s)
Humans , Apoptosis , Genetics , Ascorbic Acid , Pharmacology , Caspase 3 , Genetics , Cell Line, Tumor , Cell Proliferation , Flow Cytometry , Gene Expression Regulation, Neoplastic , Genetics , Inhibitor of Apoptosis Proteins , Microtubule-Associated Proteins , Genetics , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Objective To analyze the epidemiology,clinical and imageological characteristics,diagnosis,misdiagnosis,treatment and prognosis of pulmonary alveolar proteinosis (PAP) reported in China in recent 20 years,in an attempt to provide important clues for prompt and accurate diagnosis of PAP.Methods Clinical data of PAP from 1988 to 2008 in China were retrospectively analyzed and the clinical data of 126 patients with PAP which were misdiagnozed were summarized.Results There were 19 cases misdiagnozed with pneumonia,24 cases with pulmonary cancer,18 cases with bronchitis,19 cases with idiopathic pulmonary fibrosis,15 cases with pulmonary tuberculosis,10 cases with eosinophilic pneumonia,9 cases with sarcoidosis,5 cases with fungus pneumonia.The clinical manifestations of PAP had no specificity and the imageology manifestation was various.Its final diagnosis mainly depended on the examination of brochoalveolar lavage fluid and/or lung biopsy,and pathologic examination.Conclusions The diversity of clinical manifestations of PAP has resulted in higher clinical misdiagnosis rate.WhoLe lung irrigation is the safest and the most effective way to treat PAP.
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Epidermal growth factor receptor (EGFR) is necessary in normal epithelial cell growth. Ab-errant EGFR expression is linked with increased cell proliferation and decreased apoptosis. Mutations and over-expression of EGFR are common features of many cancers. Therefore, Therapeutic agents that target the EGFR signal pathway, such as small-molecule tyrosine kinase inhibiiors and monoclonal antibodies are now advanced in clinical development. EGFR-targeted therapies of non-small-cell lung cancer (NSCLC) have been studied as a new way.
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AIM: To investigate the therapeutic effect of chloride channel blocker tamoxifen on hypoxia pulmonary hypertension in rats.METHODS: Sixty male Wistar rats were randomly divided into therapeutic group(H/Q group),hypoxia group(H group),normal control group(C group).Mean pulmonary artery pressure(mPAP),the ratio of the weight of right ventricle to that of left ventricle plus septum[RV(LV+S)],and the ratio of the pulmonary arteriole wall area to that of vascular total area(WA/TA),were measured at 4,7,14,21 days.RESULTS:The mPAP began to increase from 7 days,and reached peak at 21 days in hypoxia group,however it is obviously lower in therapeutic group at same time.The ratio of RV(LV+S) began to increase from 14 days,and reached peak at 21 days in hypoxia group,however it is obviously lower in therapeutic group at same time.The ratio of WA/TA at 21 days in hypoxia group was significantly higher than therapeutic group,respectively(P
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Objective To evaluate the feasibility, safety, and efficacy of CT-guided permanent iodine-125 implantation for malignant tumors.Methods Thirteen lesions in 10 consecutive patients with malignant tumor were treated with CT-guided iodine-125 permanent implantation brachytherapy, of which four cases were primary unresectable carcinoma and six cases were metastases. There were 4 males and (6 females,) the mean age was 56.9 years (range 54 to 62 years). Based on the CT imaging within two weeks before the implantation of the seeds, a computer-based treatment planning system was used to determine the optimal seed distribution. Subsequent CT-guided needle placement and seed implantation were carried out. Post-implant CT scans were performed immediately and five to ten months after the implantation in all cases to assess seed distribution, complication, and curative effect. Results CT-guided iodine-125 permanent implantation was accomplished smoothly in all cases. This technique offered a better seed placement. The number of seeds implanted in one lesion was 1 to 44 (mean 18.6). No acute complications and late toxicity related to the implantation were observed. Pain relief was obtained in all four patients (100%) presenting with pain. Follow-up CT demonstrated that 3 of 13 lesions disappeared completely, eight lesions diminished, and the remaining 2 lesions had no significant change in size. Mean lesion size of pre-implant and post-implant were 3.15 cm and 2.06 cm, respectively (t=5.127, P
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Objective To discuss the role of combined external beam radiotherapy and intraluminal brachytherapy for tracheal cancer and bronchogenic carcinoma. Methods From February 1987 to August 1996, 4 patients with primary tracheal cancer, 22 with primary lung cancer and 14 (18 sites) with postoperative recurrent lung cancer were irradiated. External beam radiotherapy (EBR ) was delivered with linac X-ray to a total dose of 30-77?Gy (median 52?Gy). Intraluminal irradiation (IR ) was delivered with low dose rate 192 Ir (1.48?BGq) to a total dose of 10-53?Gy (median 28?Gy), 4-6?Gy per fraction on the bronchial mucosa. Results Complete response (CR) was obtained in 37 patients, partial response (PR) in 2 and minor response (MR) in 1. The 3-year and 5-year local control rates by Kaplan-Meier method were 75% and 65%. Twenty-three patients have survived for 3 years or longer, with a 3-year survival rate of 57.5%. Complications were rare. Conclusions Combined external beam radiotherapy and intraluminal irradiation is effective for primary lung cancer and localized tracheal cancer, possibly giving long-term survivals.
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Objective To analyze the clinical effects of internal radiation of iodine-125 seed implantation in the treatment of advanced pancreatic carcinoma.Methods Ten patients with unresectable and advanced pancreatic carcinoma admitted from Sept.2002 to June 2004 were treated by the internal and interstitial radiation using iodine-125 seed implantation. Sex:male 6, female 4. Age: median 60.8(45-81) years. Site by pre-operative CT: pancreatic head 4, body 2 and tail 4. Diameter: 6?cm 3. Pathology: all proved to be pancreatic adenocarcinoma. Staging(UICC 1997): stage Ⅲ 3, IV 7. Pain degree: Ⅲ 3,IV 7. Weight loss: median 5?kg(3-10?kg). Results All patients have been followed for 2-13 months, median 6 months. Survival of 12 months was observed in 1 patient,6 months in 3 patients, 3 months in 4 and 1 month in 2, with an average of 4.6 months. Among the 10 patients, complete response was obtained in 2, partial response in 3, no response in 4, and the other showing PD. The response rate (CR+PR) was 50%. The pain relieving rate was 60%. The seed number conformal rate was 100% by X-ray film, yet that of seed spatial distribution was only 30%. Conclusion Intra-operative internal radiation by iodine-125 seed implantation does show some therapeutic effects for advanced pancreatic carcinoma.Yet,the seed spatial distribution by seed computer treatment planning system does needs further consummate.
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T) of SNP on the APM1 gene were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay in 75 pedigrees (337 individuals) of type 2 diabetes mellitus. Genehunt software was used to analyze the transmission-disequilibrium (TDT) and to calculate SNP combining haplotypes. Meanwhile, the physiological and biochemical parameters were also determined in the pedigrees of type 2 diabetes mellitus. Results: We found no preferential transmission in the tested binding sites or any haplotypes of SNP in the APM1 gene in the filial generation of type 2 diabetes mellitus. In type 2 diabetes mellitus group, GG genotype had higher body mass index (BMI)(P=0.032) and waist circumference (WC)(P=0.030) compared to CC genotype in the patients with SNP-11377 binding site; patients with G allele had lower levels of high-density lipoprotein cholesterol (HDL-C)(P=0.006)and higher levels of fasting plasma insulin (FINS) (P=0.011) as compared to CC genotype. However, FINS levels (P=0.021) in subjects with CC genotype were significantly lower than those with G allele in healthy control group. For patients with SNP+45 binding site, those with GG genotypes had higher BMI (P=0.036) and lower levels of FINS (P=0.014) than those with TT genotypes in both groups. For patients with SNP+276 binding site, those with GG genotypes had higher BMI(P=0.043) than those with T allele in the control group. Conclusion: The SNP of the APM1 gene is not associated with the pedigrees of type 2 diabetes mellitus, but APM1 gene has influence on the function of insulin B cells and the development of obesity.