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A long-held belief is that pituitary hormones bind to their cognate receptors in classical target glands to actuate their manifold functions. However, a number of studies have shown that multiple types of pituitary hormone receptors are widely expressed in non-classical target organs. Each pituitary gland-derived hormone exhibits a wide range of nonconventional biological effects in these non-classical target organs. Herein, the extra biological functions of pituitary hormones, thyroid-stimulating hormone, follicle-stimulating hormone, luteinizing hormone, adrenocorticotrophic hormone, and prolactin when they act on non-classical organs were summarized, defined by the novel concept of an "atypical pituitary hormone-target tissue axis." This novel proposal explains the pathomechanisms of abnormal glucose and lipid metabolism, obesity, hypertension, fatty liver, and atherosclerosis while offering a more comprehensive and systematic insights into the coordinated regulation of environmental factors, genetic factors, and neuroendocrine hormones on human biological functions. The continued exploration of the physiology of the "atypical pituitary hormone-target tissue axis" could enable the identification of novel therapeutic targets for metabolic diseases.
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Humans , Pituitary Hormones/metabolism , Luteinizing Hormone , Follicle Stimulating Hormone , Prolactin , Pituitary Gland/metabolismABSTRACT
The rare endocrine and metabolic diseases, due to their varieties, face many challenges in the study of clinical diagnosis, pathogenesis, and treatment. In the past a couple of years, the research on rare endocrine and metabolic diseases has been gradually improved. The diagnosis has made great progress. The research into molecular mechanism of rare endocrine and metabolic diseases has significantly advanced. The effort in exploring the breakthroughs and progress in therapeutic methods based on the pathogenesis of the diseases has also made. This article provides a brief overview of the current status of research into diagnosis, mechanism, and treatment of rare endocrine and metabolic diseases. In addition, the article points out the problems and challenges and proposes future possibilities.
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Hypophosphatemic rickets/osteomalacia is a group of rare chronic metabolic bone diseases, which has detrimental impact on the growth, mobility, and life quality of the patients. Because of its rarity, the public awareness remains low, so does in general physician, which leads to delayed diagnosis and treatment. With a vision to standardize the diagnosis and treatment of hypophosphatemic rickets/osteomalacia, Chinese Society of Endocrinology and Chinese Society of Osteoporosis and Bone Mineral Research convened a national group of experts to compose this guideline based on current evidence, which covered the pathogenesis, diagnosis, treatment and management of hypophosphatemic rickets/osteomalacia. In summary, this work outlines recommendations for clinicians, aiming to improve the management of hypophosphatemic rickets/osteomalacia in China.
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Objective:To report a case of combined oxidative phosphorylation deficiency 28 (COXPD28) in China, identified the pathogenic mutation and explored the pathogenic mechanism preliminarily.Methods:The clinical characteristics of a patient with COXPD28 were retrospectively analyzed and the pathogenic mutations were identified by mitochondrial gene sequencing and whole exome sequencing. The wild-type and mutant plasmids of pathogenic genes were constructed, and effect of mutation on protein expression by quantitative real-time PCR (qPCR) and Western blot were evaluated. Statistical methods mainly used one-way ANOVA and LSD test.Results:A 21 year old female patient presented with lactic acid poisoning due to repeated chest distress and wheezing since childhood. The sequencing of the whole exon group gene found that solute carrier family 25 member 26 (SLC25A26) gene had a compound heterozygous mutation (c.34G>C, p.A12P; c.197C>A, p.A66E), which was the first report in China. In vitro function test showed that the expression levels of SLC25A26 mRNA and S-adenosylmethionine carrier (SAMC) protein in cells transfected with SLC25A26 mutant plasmid were significantly lower than those transfected with wild type plasmid. The p.A66E mutant plasmid reduced the expression level of SLC25A26 mRNA and SAMC protein to 6% and 26% of wild type plasmids respectively (both P<0.001), while p.A12P mutant plasmid decreased to 62% and 82% of wild type plasmids respectively ( P<0.001, P=0.044). When the double mutant (p.A66E+p.A12P) plasmids were co-transfected, the expression levels of SLC25A26 mRNA and SAMC protein decreased to 47% and 57% of the wild type plasmids, respectively ( P<0.001, P=0.001). Conclusion:The pathogenic mutation gene of this patient with COXPD28 is SLC25A26 gene mutation (p.A66E, p.A12P), which causes the decrease of SLC25A26 expression level, mitochondrial oxidative phosphorylation dysfunction, and induces COXPD28.
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To assess the correlation between thyroid function and glucolipid metabolism in type 1 diabetic adults. A retrospective analysis was conducted in 230 type 1 diabetic adults who were hospitalized in the Department of Endocrinology of Shandong Provincial Hospital Affiliated to Shandong University from January 2008 to January 2020. It showed that thyroid stimulating hormone(TSH) was significantly positively correlated with total cholesterol (TC) ( r=0.239), triglycerides (TG) ( r=0.166) and low-density lipoprotein cholesterol (LDL-C) ( r=0.249), respectively (all P<0.05). Free triiodothyronine (FT 3) was significantly negatively correlated with fasting plasma glucose (FPG) ( r=-0.272), glycated hemoglobin (HbA1c) ( r=-0.240), TC ( r=-0.197) and LDL-C ( r=-0.220), respectively (all P<0.05). Free thyroxine (FT 4) was negatively correlated with TC ( r=-0.171) and LDL-C ( r=-0.170), respectively (all P<0.05). TC was an independent predictor of TSH, FT 3 and FT 4, FT 3 and FT 4 were independent predictors of HbA1c. TSH was an independent predictor of TC, TG and LDL-C. Thyroid function is closely related to glucolipid metabolism in type 1 diabetic adults.
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In 2013,the Chinese Society of Endocrinology published the'Management ofhyperuricemia and gout:Chinese experts consensus'.In the following five years,a large number of basic researches,clinical studies,and epidemiological results had emerged and need to be popularized in order to improve the understanding and standardizing management of these conditions in general physicians.On the other hand,the methodology of developing clinical guidelines has become more and more scientific,standardized and international.Therefore,based on the 2013 version of the consensus,Chinese Society of Endocrinology decided to develop a guideline for the diagnosis and management of hyperuricemia and gout under standardized methods and procedures of evidence-based clinical practice guidelines.This article will give a review of the status of clinical practice guideline development in China and explain the methods and procedures followed when developing this guideline.We hope this work may provide some useful recommendations for other developers to draft evidence-based clinical practice guidelines.
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Objective@#To establish a mouse model of osteoarthritis (OA) and study the bone microarchitecture and bone metabolism of tibial subchondral bone in early stage of OA.@*Methods@#The mouse model of post-traumatic osteoarthritis (PTOA) with anterior cruciate ligament (ACLT) was established by using c57 mice. The Sham operation group served as the control group. All mice were fed with conventional diet. All mice were sacrificed after 4 weeks. The degeneration of knee joint was observed by HE staining and Safranin O-Fast Green staining. The number of osteoclasts was counted by TRAP staining. Micro CT was used to analyze the quantitative parameters of the microstructure of tibia subchondral bone in mice. Serum levels of bone resorption biomarker CTX I and cartilage degeneration marker CTX II were determined.@*Results@#After ACLT 4 weeks, the average score of OARSI in ACLT group was 3.2, which was higher than that in Sham group, and the joint degeneration occurred in mice, presenting the pathological characteristics of early OA. Compared with the sham operation phase, the total subchondral bone volume (TV) of ACLT group was 4.72 mm3, increased by 13.6%; the bone trabecular resolution (Tb.Sp) was 0.130 and 0.154 mm, respectively, and the ACLT group also increased by 18.8%; the bone volume/tissue volume (BV/TV) was 0.470 and 0.294, respectively, and the ACLT group decreased by 48.9%; the bone trabecular thickness (Tb.Th) was 0.162 and 0.083 mm groups, ACLT decreased by 37.5%. Trap staining showed that the number of osteoclasts per unit volume in ACLT group was 72, which was significantly higher than that in sham operation group. The CTX I of mice in the sham operated ACLT group and sham operated group were 20.9 ng/ml and 18.29 ng/ml, with an increase of 48.9% in the ACLT group; the CTX II of mice in the ACLT group and sham operated group were 35.5 ng/ml and 28.6 ng/ml, with an increase of 24.1% in the ACLT group.@*Conclusion@#ACLT Mouse model can successfully construct early OA, which confirms the early loss of osteochondral bone and the pathological changes of osteoclast activation in OA, and provides a new specific target for the treatment of OA.
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Objective To explore the relationship between different smoking status and hypothyroidism in six iodine-suitable areas of China. Methods A total of 8187 residents were investigated by cluster sampling in six cities, and 7448 residents were included in the survey. The height, weight, waist circumference, and blood pressure were measured by filling out epidemiological questionnaire. Blood samples were collected to detect thyroid stimulating hormone ( TSH) , thyroid peroxidase antibody ( TPOAb) , and thyroglobulin antibody ( TgAb) . Results The mean TSH, TPOAb, and TgAb positive rates in passive smoking and active smoking groups were all lower than those in non-smoking group ( all P<0.01) . In the active smoking group, the TSH value decreased by 0.023 units for every unit increase in smoking index. The positive rates of TgAb and TPOAb in both passive smoking and active smoking groups were lower than those in non-smoking group (all P<0.01). Active and passive smoking reduced the prevalence of hypothyroidism (both P<0.01). Among women, the risks of clinical hypothyroidism and subclinical hypothyroidism were reduced in both active and passive smoking groups. Besides, the risk of subclinical hypothyroidism decreased significantly when the smoking index was more than 70. In male population, the risk of subclinical hypothyroidism in active and passive smoking group decreased. Besides, the risk of clinical hypothyroidism and subclinical hypothyroidism decreased significantly when the smoking index was more than 70. Conclusion Smoking in iodine-suitable areas may reduce TSH level and the positive rates of TPOAb and TgAb.
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Objective To obtain a non-invasive subclinical hypothyroidism ( SCH) mouse model, and to explore microRNAs profile related to lipid metabolism in the model mice. Methods C57BL/6 male mice (8 weeks) were treated with methimazole ( MMI, 0. 08 mg/kg BW/d) to construct SCH mouse model. MicroRNAs profiling analysis was performed by real-time quantitative polymerase chain reaction ( qRT-PCR) . Results Compared with the control group, the serum thyroid stimulating hormone ( TSH ) in subclinical hypothyroidism group increased significantly (P>0.01), while the serum free thyroxine(FT4) level did not show significant difference between the two groups (P>0. 05), which was in line with the diagnostic criteria of SCH. SCH mice was accompanied by dyslipidemia and liver lipid metabolism disorders. Four lipid metabolism related miRNAs, miR-33, miR-122, miR-199a-5p, and miR-375 in the liver of SCH mice were significantly decreased compared with those of control ( P>0. 05). Conclusion The noninvasive SCH model generated by MMI and miRNAs profile provide an animal model and a molecular basis for the study of SCH related lipid metabolism disorders.
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OBJECTIVE: To analyze the data of occupational pneumoconiosis caused by bentonite dust. METHODS: The data of 2 cases of occupational pneumoconiosis caused by bentonite dust was retrospectively analyzed. RESULTS: Both 2 cases had a clear history of occupational exposure to bentonite dust. Case 1 has a 14 years and case 2 has 7 years of bentonite exposure history. The radiographic examination of chest X-ray in both cases showed diffuse miliary nodules with high density in both lungs. The small opacity total profusion of chest X-ray images in both cases belong to category 2. The small opacity of lung in case 1 distributed in 5 areas,while case 2 distributed in 4 areas. The pulmonary function test for case 1 showed that the forced vital capacity( FVC) was 81. 4%,the first second forced expiratory volume( FEV_1) was 76. 8%and FEV_1/FVC was 74. 4%,mild pulmonary ventilation damage; lung function test results for case 2 showed: FVC was84. 0%,FEV_1 was 90. 0%,FEV_1/FVC was 93. 0%,pulmonary ventilation function was in the normal range. Case 1 was diagnosed as occupational pneumoconiosis stage Ⅱ( bentonite); Case 2 was diagnosed as occupational pneumoconiosis stageⅠ( bentonite). CONCLUSION: The clinical symptoms and signs of occupational pneumoconiosis caused by bentonite dust are not obvious. Its diagnosis is mainly based on X-ray chest radiograph.
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Objective@#To explore window settings technology in the digital radiography of pneumoconiosis, and to improve the quality of the digital chest radiographs.@*Methods@#25 female workers in a asbestos product processing enterprise were examined with high-kV and DR chest radiographies on same day. Consistency of pneumoconiosis diagnosis@*results@#were assessed. Results In the 75 groups of radiograph quality results, there are 62 groups that need to be processed window in order to make the correct diagnosis. The crude agreement among two chest radiographs was 28% for the profusion of small opacities and 76% for the classification of pneumoconiosis, the weighted kappa value (κ) was 0.33 (95%CI: 0.12-0.54) and 0.67 (95%CI: 0.42-0.92) .@*Conclusions@#window settings technology can display different parts of chest and make DR chest radiograph to meeting the requirements of chest radiograph quality for pneumoconiosis.
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Objective@#To observe the therapeutic response of radiosynovectomy with p-32 colloid on hemophilic arthropathy, and to assess the effects of radiosynovectomy with Denver Score on hemophilic arthritis staging.@*Methods@#Radiosynovectomy with p-32 colloid was performed on 326 hemophilic arthritis patients (405 joints) , and recorded bleeding before and after treatment. The MRI performance of 102 joints was evaluated by using Denver scoring system, then was divided into 0-6 and 7-10 groups. Finally, the differences between 2 groups were analyzed.@*Results@#Average pain score of all hemophilic arthritis patients at 6 months, 1, 2 and 3 years post treatment decreased from 3.2±2.4 (n=326) to 1.2±0.6 (n=285, P=0.021) , 1.7±0.5 (n=242, P=0.032) , 2.1±1.1 (n=212, P=0.030) and 2.2±1.6 (n=176, P=0.037) , respectively. The frequency of bleeding in 405 joints at 1, 2 and 3 years post treatment decreased from 15.1±3.6 to 2.1±0.7, 4.3±0.6, and 4.8±0.8 times per year (P<0.01) , respectively; Meanwhile, the proportions of significantly ameliorated joints’ activities were observed as of 68.50% (248/362) , 58.39% (181/310) , 55.67% (162/239) and 42.61% (75/176) , respectively. The frequencies of haemarthrosis at 1 and 2 years post treatment in patients with 0-6 Denver Score (45 target joints) reduced from 13.0±1.9 to 1.3±0.6 (P=0.002) and 3.1±0.9 (P=0.009) times per year, respectively, which also decreased in 7-10 group (57 target joints) from 16.6±2.1 to 3.1±0.9 (P=0.008) and 5.7±1.1 (P=0.004) times per year, respectively. There was no statistical difference between 0-6 and 7-10 groups before treatment in the terms of haemarthrosis frequency (P=0.773) . However, 7-10 group had higher haemarthrosis frequency at 1 and 2 years post treatment compare with 0-6 group (P=0.028 and 0.042, respectively) . Synovial volumes in 29 joints reduced after 6 month when compared with baseline [ (2 362.15±32.41) mm3 vs (3 012.40±39.78) mm3, t=7.621, P<0.001].@*Conclusion@#Radiosynovectomy with p-32 colloid on haemophilic synovitis was a safe and effective procedure. The patients with Denver Score of 0-7 had lower frequency of haemarthrosis.
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Objective To compare the suture bridge (SB) and conventional double-row (DR) suture in the repair of full-thickness rotator cuff tear.Methods From May 2013 through July 2016,48 patients with full-thickness rotator cuff tear were randomly divided into 2 even groups (n =24).SB group was repaired using the SB technique while DR group using conventional DR technique.The 2 groups were compared in terms of operation time,postoperative shoulder function scoring and incidence of re-tear.Results All the patients were followed up for 8 to 24 months (average,15.3 months).In DR group,the visual analogue scale (VAS) scores decreased from preoperative 6.3 ± 2.3 to 1.0 ± 0.4 at the last follow-up,the America Shoulder and Elbow Surgeons (ASES) scores increased from preoperative 49.3 ± 8.5 to 90.0 ± 2.5 at the last follow-up,and the Constant scores increased from preoperative 58.7 ± 12.5 to 88.1 ± 4.0 at the last follow-up.In SB group,the VAS scores decreased from preoperative 6.0 ± 1.9 to 0.9 ± 0.8 at the last follow-up,the ASES scores increased from preoperative 50.2 ± 6.2 to 89.5 ± 3.4 at the last follow-up,and the Constant scores increased from preoperative 57.3 ± 7.5 to 90.0 ± 3.2 at the last follow-up.All the comparisons showed a significant difference between preoperation and the last follow-up (P ＜ 0.05),but an insignificant difference between the 2 groups (P ＞ 0.05).SB group used significantly less operation time (74.5 ± 19.0 min) than DR group (86.5 ± 21.0 min),and reported significantly lower incidence of re-tear (4.2％) than DR group (25.0％) (P ＜ 0.05).Conclusions In arthroscopic repair of full-thickness rotator cuff tear,SB technique shows few therapeutic advantages over conventional DR technique,but the former needs less operation time and leads to lower incidence of re-tear.
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Objective To compare the suture bridge (SB) and conventional double-row (DR) suture in the repair of full-thickness rotator cuff tear.Methods From May 2013 through July 2016,48 patients with full-thickness rotator cuff tear were randomly divided into 2 even groups (n =24).SB group was repaired using the SB technique while DR group using conventional DR technique.The 2 groups were compared in terms of operation time,postoperative shoulder function scoring and incidence of re-tear.Results All the patients were followed up for 8 to 24 months (average,15.3 months).In DR group,the visual analogue scale (VAS) scores decreased from preoperative 6.3 ± 2.3 to 1.0 ± 0.4 at the last follow-up,the America Shoulder and Elbow Surgeons (ASES) scores increased from preoperative 49.3 ± 8.5 to 90.0 ± 2.5 at the last follow-up,and the Constant scores increased from preoperative 58.7 ± 12.5 to 88.1 ± 4.0 at the last follow-up.In SB group,the VAS scores decreased from preoperative 6.0 ± 1.9 to 0.9 ± 0.8 at the last follow-up,the ASES scores increased from preoperative 50.2 ± 6.2 to 89.5 ± 3.4 at the last follow-up,and the Constant scores increased from preoperative 57.3 ± 7.5 to 90.0 ± 3.2 at the last follow-up.All the comparisons showed a significant difference between preoperation and the last follow-up (P ＜ 0.05),but an insignificant difference between the 2 groups (P ＞ 0.05).SB group used significantly less operation time (74.5 ± 19.0 min) than DR group (86.5 ± 21.0 min),and reported significantly lower incidence of re-tear (4.2％) than DR group (25.0％) (P ＜ 0.05).Conclusions In arthroscopic repair of full-thickness rotator cuff tear,SB technique shows few therapeutic advantages over conventional DR technique,but the former needs less operation time and leads to lower incidence of re-tear.
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Objective To detect the levels of anti-measles antibody in a healthy population in Jin-shan District of Shanghai and to provide references for making a better immunization strategy against measles in this area. Methods Stratified sampling was used to collect 390 serum sample in 12 age groups with an equal number of males and females as well as resident and floating population. Indirect enzyme-linked immu-nosorbent assay (ELISA) was performed to detect the concentrations of IgG antibody against measles in dif-ferent groups. Results The overall positive rate and protective rate of anti-measles antibody were 78.21% and 41.28%, respectively. No significant difference in the positive rates of antibody was found between males and females, or resident and floating population (P>0.05). Antibody levels varied significantly among different age groups(χ2=191.214,P<0.01). More than 80.00% of the people aged≥1 year were positive for anti-measles antibody and the positive rates in children 3 to 6 years old and people aged ≥40 years reached 100% and over 90.00%,respectively. Infants under 8 months of age,having the lowest posi-tive rate of antibody(10.00%),were susceptible to measles. The highest protective rate was found in chil-dren aged 8 months to 9 years,followed by that in people≥40 years. Levels of anti-measles antibody in peo-ple aged 10 to 39 years mainly ranged from 200 mIU/ml to 800 mIU/ml. No significant difference in anti-body levels was found among people aged≥8 months who had different immunization histories(χ2=1.140,P>0.05). Conclusion The average level of anti-measles antibody was high in Jinshan District in 2015,in-dicating that the people in that area had a relatively high immunity to measles,but the positive rate of anti-body needed to be further improved. Infants under 8 months old and people 10 to 39 years old were the main susceptible groups. It is suggested that measles vaccination efforts should be focused on susceptible popula-tion and women of childbearing age to improve herd immunity.
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[Summary] Glucose-dependent insulinotropic polypeptide ( GIP) is known to promote the release of insulin fromβcells, but this function suffers from a defect in obese patients. More and more evidences suggest that GIP is the bridge between a high-fat diet, obesity, insulin resistance, and type 2 diabetes. With the deepening of research in GIP in type 2 diabetes and obesity, GIP may provide new ideas for the treatment of type 2 diabetes and obesity.
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Objective To understand the characteristics of common causes of acute pancreatitis (AP) in China and evaluate the association of the aetiology with gender and mortality.Methods The relevant literature was searched by the China journal full-text database (CNKI),Wanfang database,Weipu database and other databases and proceeding.Based on collecting data retrieval strategy,according to the inclusion criteria selection literature,Meta analysis was performed mainly for gallstone,hyperlipidemia,alcohol and other AP from the aspects of gender and case fatality rate.Results The Meta analysis included 11 articles which were accordance with the criteria,involving 13 601 patients,including 6 732 cases of biliary AP,1 372 cases of hyperlipidemia AP and 1 169 cases of alcohol AP.The Meta analysis showed that biliary AP male to female ratio was 0.79 ∶ 1,hyperlipidemic AP male to female ratio was 1.54 ∶ 1,alcoholic AP male to female ratio was 10.47 ∶ 1,overeating AP male to female ratio was 1.29 ∶ 1,and there was significant difference (P ＜ 0.05).Alcoholic AP mortality rate was the highest,which was 2.81 times than the biliary AP and 2.46 times than the hyperlipemic AP,and there was significant difference (P ＜0.05).Conclusions Biliary tract disease is the main etiologic cause of AP in China,and there are more females than males.The mortality rate of alcoholic pancreatitis is the highest,and there are more males.But we should investigate further high-quality,large-scale trails in patients with AP.
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Objective To investigate the ability of induction of specific cytotoxic T lymphocytes (CTL) stimulated by dendritic cells (DCs) co-transfected with MUC1 and survivin mRNA of human pancreatic cancer,and to provide the experimental basis for the treatment of human pancreatic cancer with multi-epitope DC vaccine.Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs) of 6 patients with pancreatic cancer.Human pancreatic cancer cell line MiaPaCa-2 was routinely cultured,after being transcripted and amplified by RT-PCR,MUC1 and survivin mRNA were co-transfected or individually transfected into DCs by electroporation,and they were named as DC-MUC1,DC-survivin,DC-MUC1 + survivin.The expression of MUC1 and survivin mRNA in DCs were detected by real-time PCR.The survival rate of transfected DCs were determined by MTT method.The lymphocyte proliferation ability was evaluated by mixed cell culture method.The Th1 cytokine releasing of antigen-specific CTLs were measured by ELISA assay.Results Mature DCs were obtained,the positive expression rates of surface markers CD40,HLA-DR,CD83 and CD86 were 34.31％,50.21％,89.17％ and 73.62％,respectively.The expression amount of MUC1 mRNA of DC-MUC1 was 36.24 ± 5.17,and the expression amount of survivin mRNA of DC-survivin was 34.53 ± 4.02,while the expression amounts of MUC1,survivin mRNA of DC-MUC1 + surviving were 31.79 ±4.26 and 14.67 ± 2.96,which were significantly lower than that in individual transfection group (P ＜ 0.05).The survival rate of DC-MUC1 + surviving was decreased in a time dependent manner,which was significantly lower than that in individual transfection group (about 50.21％ vs 80％ at 24 h,P ＜0.05).When DC/T cells ratio was 1∶ 10,1∶ 20,the autologous T cell proliferation index of MUC1 and survivin mRNA in co-transfection DC group was significantly higher than that in individual transfection group (P ＜ 0.05) ;when DC/T cells ratio was 1∶ 40,1∶ 80,the difference of proliferation index was not statistically significant.When DC/T cells ratio was 1∶ 10,after 14 d culture,the expressions of IL-2 in DC-MUC1,DC-survivin,DC-MUC1 + surviving were (892.73 ± 32.9),(713.62 ± 56.37),(1884.37 ± 95.21) pg/ml,and the expressions of granzyme B were (501.62 ± 12.30),(203.84 ± 12.55),(1193.15 ± 86.04) pg/ml ; and the expressions of IFN-γ were (981.50 ± 47.82),(696.05 ± 41.66),(2237.94 ± 189.55) pg/mL.The corresponding values in DC-MUC1 + surviving group were significantly higher than those in individual transfection group (P ＜ 0.05) ; while the difference of IL-10 was not statistically significant.Conclusions DCs co-transfected with MUC1 and survivin mRNA have a stronger ability to stimulate specific CTL in vitro than individual antigen loaded DCs.
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Objective To investigate the anti-tumor immune response induced by human pancreatic cancer mucin 4 mRNA and human telomerase reverse transcriptase (hTERT) mRNA cotransfected dendritic cells (DC),and to provide the experimental evidences for the treatment of pancreatic cancer with multi-epitope loaded DC vaccine.Methods DC were isolated from peripheral blood mononuclear cells (PBMC) of six patients with HLA-A2+ pancreatic cancer and cultured.Mucin 4 mRNA and hTERT mRNA were transcripted and amplified in vitro,which were transfected into DC separately or in order by eleetroporation.DC were cultured for 48 hours.The expressions of mucin 4 and hTERT in DC were detected by quantitative real-time polymerase chain reaction (PCR) and Western blot.The survival rates of transfected DC were determined by methylthiazolyl tetrazolium (MTT) method.The cytotoxic T lymphocyte (CTL) activation induced by mucin 4 mRNA and hTERT mRNA transfected DC were evaluated by interferon (IFN)-γ release assays (enzyme linked immunosorbent assay) method.The cytotoxicity of CTL induced by mucin 4 mRNA and hTERT mRNA transfected DC in pancreatic cancer cell lines MiaPaCa-2,Capan-2,AsPC-1 and Pane-1 was measured by 51Cr standard cytotoxicity test.Student t test was performed for statistical analysis.Results After in order transfection of mucin 4 mRNA and hTERT mRNA for 48 h,the relative quantity of the expression of mucin 4 and hTERT in DC were 30.09±5.24 and 12.87±3.36,which were lower than the relative quantity of the expression in DC transfected separately (38.54±6.21 and 36.35±5.03,t=3.469,6.721,both P＜0.05).After transfected in order for 96 hours,the survival rate of DC decreased to 52.17％,which was lower than that of DC transfected separately (around 80％).The quantity of IFN-γ releasing of specific CTL induced by mucin 4 mRNA and hTERT mRNA cotransfected DC was (32.57±2.01) U/mL in 24 hours,which was higher than that of CTL induced by DC transfected with mucin 4 mRNA ((23.06±4.74) U/mL) or hTERT mRNA ((16.82±3.67) U/mL) separately (1=5.092,7.141,both P＜0.05).After co-transfected with mucin4 mRNA and hTERT mRNA,DC could effectivly induce HLA-A2+/mucin 4+/hTERT+ specific CTL immune responses,however there was no significant cytotoxicity in HLA-A2+ pancreatic cancer cells.Conclusion The induction of CTL anti-tumor immune response by DC co-transfected with mucin4 mRNA and hTERT mRNA is more significant compared with that by single antigen loaded DC.
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Objective To investigate the effects of PTEN and KAI1 double gene transfection on proliferation,metastasis in AsPC1 pancreatic cancer cells under hypoxic condition.MethodsRecombinant vectors that over expressing PTEN and KAI1 protein which was established previously were double transfected into hypoxic AsPC1 cells.Western blot was performed to measure the expression level of PTEN and KAI1.Then,cell proliferation was detected by MTT,and colony forming assay were used to test the ability of tumor cells forming colonies,and transwell assay was used to evaluate metastatic function.ResultsAfter double gene trnsfection,both PTEN and KAI1 protein expression was significantly up-regulated,which was 2.05 and 1.5 1 folds higher than that of empty vector group; and cell proliferation of hypoxic AsPC1 cells was suppressed significantly (0.5 vs 0.8,P =0.00031 ),number of colony formation was significantly decreased [ (41.67 ± 5.03) vs (86.00 ± 7.81 ),P =0.017) ] ; the capacity of metastasis was significantly decreased (0.68 ± 0.05 vs 1.23 ± 0.03,P =0.0025 ).ConclusionsDouble gene transfection of PTEN and KAI1 could inhibit proliferation and metastatic activity of hypoxic AsPC1 cells,which might indicate that combined gene therapy may play a role in the treatment of pancreatic cancer.