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OBJECTIVE@#To analyze variants of TSC1 and TSC2 genes in a Chinese patient with tuberous sclerosis complex (TSC).@*METHODS@#Peripheral blood samples were collected from the patient and her parents with informed consent. Following extraction of genomic DNA, potential variants of the TSC1 and TSC2 genes was detected by using targeted capture next-generation sequencing (NGS) and Sanger sequencing.@*RESULTS@#The patient was found to harbor a de novo mosaicism variant c.3295_3298delG (Val1100CysfsTer3) of the TSC2 gene, with the proportion of the mutant allele determined as 13.4%, which was confirmed by Sanger sequencing. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.3295_3298delG (Val1100CysfsTer3) variant was predicted to be pathogenic (PVS1+PS2+PM2).@*CONCLUSION@#The mosaicism heterozygous variant of c.3295_3298delG of the TSC2 gene, as detected by both NGS and Sanger sequencing, probably underlay the TSC2 in this patient.
Subject(s)
Female , Humans , Mosaicism , Mutation , Tuberous Sclerosis/genetics , Tuberous Sclerosis Complex 1 Protein/genetics , Tuberous Sclerosis Complex 2 Protein/geneticsABSTRACT
In recent years, the incidence of hematological malignancies has increased year by year, although with the progress of chemotherapy, targeted therapy and hematopoietic stem cell transplantation, the efficacy of some types of lymphoma and leukemia has been greatly improved.However, the prognosis is still poor and the fatality rate is high.Finding new strategies to improve the curative effect has become an urgent problem to be solved.Oncolytic virus can promote anti-tumor response by selectively destroying tumor cells and inducing specific anti-tumor immune response.At present, oncolytic virus has become an effective strategy for the treatment of a variety of malignant tumors.A lot of breakthrough progress has also been made in the treatment of hematological malignant tumors.In this paper, the latest research progress of oncolytic virus in the treatment of hematological malignant tumors in recent years is reviewed.
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Objective@#To explore the prognostic relationship between initial absolute lymphocyte count(ALC) of peripheral blood and primary immune thrombocytopenia (ITP) in children, in order to provide basis for judging the prognosis and treatment of ITP in children.@*Methods@#Clinical data of 166 children with primary ITP in children admi-tted at the Affiliated Hospital of Guizhou Medical University from January 2014 to March 2018 were analyzed retrospectively, and they were followed up by clinic and telephone, the prognostic factors (gender, age, ethnicity, inducement, bleeding, initial ALC, platelet count and treatment) were statistically analyzed, and the relationship between ALC of peripheral blood and the prognosis of children with ITP was observed.@*Results@#Of 166 children with ITP, 89 cases (53.6%) had remission within 3 months, 18 cases (10.8%) within 3-12 months, 20 cases (12.0%) within 1-4 years, a total of 39 cases (23.5%) were refractory (no remission in 1-10 years), the remission rate within 1 year was 64.5%, the total remission rate was 76.4%.ALC of remission cases was (4.58±2.87)×109/L within 3 months, (4.47±2.04)×109/L within 3-12 months, and (2.86±1.61)×109/L within 1- 4 years.Thirty-nine cases (23.5%) were refractory(no remission in 1-10 years), ALC of them was (2.07±0.98)×109/L, and there were significant differences among different groups (F= 12.06, P<0.01). Univariate analysis showed that age (F=27.28), pre-morbidity history of infection and vaccination history(χ2= 9.31), initial ALC (F=12.06) at initial diagnosis were related with the prognosis of children with ITP (all P<0.05). Multivariate analysis showed that the history of infection before onset, the history of vaccination(95%CI: 0.19 to 1.51, P<0.05)and initial ALC(95%CI: -0.64 to -0.23, P<0.001) at the time of initial diagnosis were independent factors affecting the prognosis of children with ITP.The receivers operating characteristic (ROC) was drawn with the development of chronic disease (course >12 months) as state variable and ALC as test variable.The area under curve(AUC) was 0.765(P<0.05), cut-off point was 3.925×109/L, sensitivity was 0.542, and specificity was 0.966.@*Conclusions@#Initial ALC can be used as one of the prognosis index of ITP in children, and low ALC is the risk signal for chronic development of ITP in children.
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Objective To investigate the impact of the changes of posttreatment karnofsky performance status (KPSpost) on the overall survival (OS) for patients with stage Ⅳ non-small cell lung cancer (NSCLC) underwent concurrent chemoradiation.Methods A total of 279 patients (male 198 and female 81) with histological confirmed stage Ⅳ NSCLC were enrolled in this study with a median age of 58 years old (range 22 to 80 years old).There were 166 cases of squamous carcinoma,87 cases of adenocarcinoma,and 22 cases of unclassified carcinoma,respectively.All enrolled patients received more than 2 cycles of chemotherapy and more than 36 Gy of concurrent radiotherapy.Kaplan-Meier method and Log-rank test were applied to evaluate OS.Multivariate analyses were carried out by the Cox proportionalhazard model.Chi-square test and logistic regression analysis were used to explore the related factors of KPSpost.Results There were 198 patients with improved KPSpost and 81 patients with decreased KPSpost,respectively.Univariate and multivariate analyses indicated that the improvement of KPSpost was associated with longer OS.Logistic regression analysis showed that the improvement of KPSpost was positively related with treatment of more than 4-6 cycles chemotherapy concurrent with over 63 Gy radiation to primary tumor.The improvement of KPSpost also correlated positively with disease control rate (DCR),but negatively with PLT toxicity and radiation esophagitis.Conclusions KPSpost was an independent prognostic factor of OS for patients with stage Ⅳ NSCLC underwent concurrent chemoradiation.Chemotherapy of 4-6 cycles and concurrent over 63 Gy radiotherapy dose to primary tumor,as well as DCR were positive factors for KPSpost improvement.However,stage 3-4 PLT toxicities and radiation esophagitis decreased the KPSpost.
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In recent years,the incidence of hematological malignancies has increased year by year,although with the progress of chemotherapy,targeted therapy and hematopoietic stem cell transplantation,the efficacy of some types of lymphoma and leukemia has been greatly improved.However,the prognosis is still poor and the fatality rate is high.Finding new strategies to improve the curative effect has become an urgent problem to be solved.Oncolytic virus can promote anti-tumor response by selectively destroying tumor cells and inducing specific anti-tumor immune response.At present,oncolytic virus has become an effective strategy for the treatment of a variety of malignant tumors.A lot of breakthrough progress has also been made in the treatment of hematological malignant tumors.In this paper,the latest research progress of oncolytic virus in the treatment of hematological malignant tumors in recent years is reviewed.
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Objective To explore the prognostic relationship between initial absolute lymphocyte count(ALC) of peripheral blood and primary immune thrombocytopenia(ITP)in children,in order to provide basis for judging the prognosis and treatment of ITP in children. Methods Clinical data of 166 children with primary ITP in children admi_tted at the Affiliated Hospital of xuizhou medical University from January 2014 to march 2018 were analyzed retrospec_tively,and they were followed up by clinic and telephone,the prognostic factors( gender,age,ethnicity,inducement, bleeding,initial ALC,platelet count and treatment)were statistically analyzed,and the relationship between ALC of pe_ripheral blood and the prognosis of children with ITP was observed. Results Of 166 children with ITP,89 cases (53. 6%)had remission within 3 months,18 cases(10. 8%)within 3_12 months,20 cases(12. 0%)within 1_4 years,a total of 39 cases(23. 5%)were refractory(no remission in 1_10 years),the remission rate within 1 year was 64. 5%,the total remission rate was 76. 4%. ALC of remission cases was(4. 58 ± 2. 87)×109/L within 3 months, (4. 47 ± 2. 04)×109/L within 3 _12 months,and(2. 86 ± 1. 61)×109/L within 1 _ 4 years. Thirty_nine cases (23. 5%)were refractory(no remission in 1_10 years),ALC of them was(2. 07 ± 0. 98)×109/L,and there were significant differences among different groups( F = 12. 06,P <0. 01 ). Univariate analysis showed that age( F =27. 28),pre_morbidity history of infection and vaccination history(χ2 = 9. 31),initial ALC( F=12. 06)at initial diagnosis were related with the prognosis of children with ITP(all P<0. 05). multivariate analysis showed that the his_tory of infection before onset,the history of vaccination(95%CI:0. 19 to 1. 51,P <0. 05)and initial ALC(95%CI:_0. 64 to _0. 23,P<0. 001)at the time of initial diagnosis were independent factors affecting the prognosis of children with ITP. The receivers operating characteristic(ROC)was drawn with the development of chronic disease(course >12 months)as state variable and ALC as test variable. The area under curve(AUC)was 0. 765(P<0. 05),cut_off point was 3. 925×109/L,sensitivity was 0. 542,and specificity was 0. 966. Conclusions Initial ALC can be used as one of the prognosis index of ITP in children,and low ALC is the risk signal for chronic development of ITP in children.
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Objective To analyze the survival and toxicity after concurrent chemoradiotherapy in patients of different ages with stage Ⅳ non-small cell lung cancer (NSCLC).Methods Clinical data of 282 NSCLC patients in two prospective studies were retrospectively analyzed,who completed the protocol (at least 2 cycles of chemotherapy and thoracic radiation doses of ≥36 Gy).Among them,44 patients were assigned into in the young group (≤ 45 years old),161 patients in the middle-age group (46-64 years old) and 77 patients in the elderly group (≥ 65 years old).The clinical characteristics of patients among different groups were analyzed by x2 test.The overall survival (OS) was calculated by Kaplan-Meier method.Stratified analysis was performed by Log-rank test.Multi-factor prognosis analysis was conducted by Cox's proportional hazards regression model.Results The incidence of NSCLC in the male patients in the elderly group was higher than that in the middle-age and young groups.The 1-,2-,3-and 5-year OS did not significantly differ among different groups (P=0.810).The OS did not significantly differ among patients of the same gender,pathological type,T stage,N stage,metastasis status,same chemotherapy cycle,primary tumor dose and comprehensive treatment and short-term response (all P＞0.05).The incidence of adverse events did not considerably differ among different groups.Multivariate analysis demonstrated that age was not an independent factor for survival (P＞ O.05).Conclusion Patients of different ages with stage Ⅳ NSCLC obtain similar survival benefits and adverse events after concurrent chemoradiotherapy.
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The efficacy of palliative care was definitive with two-dimensional radiotherapy for stage IV non-small cell lung cancer(NSCLC).However,theimpact of radiotherapy on survival was not well indicated,and some study resultsindicating prolonged survival were not accepted. Along with the advancesin three-dimensional radiotherapy (3DRT),wide application of comprehensive treatment,and the understanding of the association between different metastatic status and survival,prospective and retrospective studies have demonstrated that chemotherapy combined with 3DRT for primary tumor is more effective than chemoradiotherapy alone in improving symptoms and prolonging survival,especially for oligometastases of stage IV NSCLC.The dose to primary tumor is closely related to survival,and high-dose radiotherapy may be more likely to prolong survival. Further studies,however,areneeded to take into accountproblems such as thedose,timing,and technical selection of radiotherapy.
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OBJECTIVE@#To investigate the rapid and accurate method for the detection of dengue virus (DENV) by using nicking enzyme assisted strand-displacement amplification (SDA) combined with gold nanoparticles-based lateral flow strip.@*METHODS@#Total RNA of the virus was extracted by using magnetic beads method and transcribed to cDNA for SDA detection system. Nicking enzyme-assisted method was used for detecting DENV, and agarose gel electrophoresis was used for analyzing the sensitivity of SDA amplification products. A gold nanoparticles-based lateral flow strip was developed based on the principle of nucleic acid base complementary pairing to design the test line and control line. The gold particles were prepared by using sodium citrate reduction method for gold nanoparticles-based lateral flow strip construction.@*RESULTS@#The sensitivity of the SDA method was 10 fmol/L, and the sensitivity of gold nanoparticles-based lateral flow strip based on SDA method was also 10 fmol/L. In a linear range from 10 fmol/L to 10 fmol/L, the corresponding linear correlation coefficient () of DENV was 0.98. The specificity of nanoparticles-based lateral flow strip based on SDA for DENV detection was high, which was no crossing with other control groups.@*CONCLUSIONS@#A gold nanoparticles-based lateral flow strip based on SDA method for DENV detection has been established, which is convenient, fast, and the result is visible to naked eyes.
Subject(s)
Humans , Dengue , Diagnosis , Dengue Virus , Gold , Metal Nanoparticles , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
Objective To investigate the efficacy,adverse reactions and immune function of time-adjusted chemotherapy combined with intensity-modulated radiation therapy (IMRT) and conventional chemotherapy combined with IMRT for locally advanced nasopharyngeal carcinoma.Methods Random number grouping method was used to divide 66 cases of locally advanced nasopharyngeal carcinoma into 2 groups,of which 36 cases in the time-adjusted chemotherapy group and 30 cases in the conventional group.Both of them received docetaxel + cisplatin + fluorouracil regimen to induce chemotherapy for 2 cycles.The time-adjusted chemotherapy group was treated with intravenous injection of electronic automatic injection pump,the conventional group was treated with conventional intravenous infusion,and both groups were treated with synchronous cisplatin combined with IMRT.Calculated survival rate was generated by Kaplan-Meier method and long-term adverse reactions was evaluated according to CTC 3.0 criteria.Results The 3-year overall survival (OS) rate was 86.1％ and 93.3％ in the time-adjusted chemotherapy group and the regular group,the 3-year progress-free survival (PFS) was 83.3％ and 93.3％,the 3-year RFS was 88.5％ and 93.3％,and the 3-year recurrence-free survival was 94.1％ and 100％ respectively with no statistically significant difference (P ＞ 0.05).The dryness and hearing loss of the time-adjusted chemotherapy group had a decreasing trend compared with the conventional group.However,CD3 +,CD3 + CD4 +,CD3 + CD4 + CD8 +,and CD4 +/CD8 + of the time-adjusted chemotherapy group had an increasing trend compared with the conventional group.Conclusions Both time-adjusted chemotherapy and conventional chemotherapy combined with IMRT had comparable mid-term efficacy,but the former had lower adverse reactions,improved quality of life and immune function.Trial registration Chinese clinical trial registry,ChiCTR1800016809
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Objective To evaluate the differences of toxicities,therapeutic efficacy and immune function between induction chemotherapy followed by sinusoidal chrono-modulated infusion and flat intermittent infusion of cisplatin (DDP)with intensity-modulated radiotherapy (IMRT) in patients with locoregionally advanced nasopharyngeal carcinoma (NPC).Methods Seventy patients with biopsydiagnosed stages Ⅲ and Ⅳ B NPC (according to the 2010 UICC staging system) were treated with two-cycle induction chemotherapy before chemoradiotherapy in Guizhou Cancer Hospital.The TPF chemotherapy regimen was administered as follows:The TXT and DDP with the dose of 75 mg/m2 was carried out by bolus infusing for the first day,the 5-FU with 750 mg · m-2 · d-1 was carried out by continuous intravenous pumping for the first day to fifth day(120 h).The induction chemotherapy was 21 days per cycle,for two cycles.After that all patients were randomly treated with 2-3 cycles of sinusoidal chronomodulated infusion or flat intermittent constant rate infusion of DDP with IMRT.Using a multi-channel programmed pump,the patients were given 12 h continuous infusions of DDP (100 mg/m2) for day one,repeated every 3 weeks for 2-3 cycles.DDP was administered from 10:00 am to 10:00 pm.Concurrent radiotherapy regimen was administered as follows:GTVnx 69.96-73.92 Gy/33 f,PTVnx 69.96 Gy/33 f,PTVnd 69.96 Gy/33 f,PTV1 60.06 Gy/33 f,PTV2 50.96 Gy/28 f.Results The main toxicities of chemoradiotherapy in the group of sinusoidal chrono-modulated infusion were bone marrow suppression:leukocytes,and then nausea,oral mucositis and hemoglobin.The main toxicities of chemoradiotherapy in the group of flat intermittent constant rate infusion were bone marrow suppression:hemoglobin,leukocytes,and then nausea,oral mucositis.No significant differences were observed for toxicities(P ＞ 0.05).After concurrent chemoradiotherapy,the complete response rate (CR),partial response rate (PR),stable disease rate(SD),progressive disease rate (PD) and overall response rate (ORR) were 11.4％,85.7％,2.9％,0 and 97.1％ in the group of sinusoidal chrono-modulated infusion.The CR,PR,SD,PD,ORR in the group of flat intermittent constant rate infusion were 22.9％,74.2％,2.9％,0,97.1％,respectively.However,there was no significant differences of effect in the two Arms (P ＞ 0.05).For sinusoidal ehrono-modulated infusion and flat intermittent infusion group,the 2-year overall survival(OS) were 82.9％ and 94.3％ respectively,the 2-year progression-free survival(PFS) were 77.1％,91.4％ respectively,and the 2-year distant metastasis free survival (DMFS) were 82.9％,91.4％ respectively.The value of CD3 + in the group of sinusoidal chrono-modulated infusion was higher than the group of flat intermittent constant rate infusion after therapy (Z =3.254,P ＜ 0.05).The value of CD4 +,CD8 +,CD16 + CD56 +,CD19 +,and CD4 +/CD8 + had no differences in two Arms (P ＞ 0.05).Conclusions No significance differences on the toxicities,therapeutic efficacy and survival were observed between the two groups,but immune function might be improved in the sinusoidal chrono-modulated infusion group.
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Objective: To investigate the nutritional status of children acute lymphoblastic leukemia (ALL) during induced remission stage. The effects of the disease itself, treatment, and complications of malnutrition were all analyzed. Methods: From the medical re-cords of children with ALL in the pediatric hematological department of the Affiliated Hospital of Guizhou Medical University, we col-lected basic information of the children; monitored the height and weight of the children on the first, 15th, and 33rd days of induced remission treatment; and calculated their body mass index (BMI), as well as results of laboratory tests such as serum albumin and se-rum prealbumin. SPSS 23.0 software was used for data analysis. Results: In 40 children with ALL, there were 16 cases of malnutrition on the first day of induced remission treatment accounting for 40%, 14 cases on both the 15th day and the 33rd day of treatment ac-counting for 35%. Among children with malnutrition, 4 cases of moderate to severe malnutrition occurred on the first day of treat-ment accounting for 25.0%, 9 cases on the 15th day accounting for 64.3%, and 12 cases on the 33rd day accounting for 85.7%. Com-pared with day 1 and day 33, the difference was statistically significant (P<0.017). In the induced remission period, the BMI on the first day was (15.98±2.17) kg/m2; on the 15th day, it was (15.65±2.20) kg/m2; and on the 33rd day, it was (15.66±1.92) kg/m2. The differ-ence between the three was not statistically significant (P=0.730). In the single-factor analysis of factors related to the nutritional sta-tus of children, infection, digestive system involvement, and serum albumin levels were related to the occurrence of malnutrition, and we performed multifactor analysis of these three factors. The difference between the level of infection and serum albumin and the oc-currence of malnutrition was statistically significant (P<0.05). Conclusions: During the induced remission, the malnutrition degree of some children with ALL was aggravated. Infection was a high-risk factor for malnutrition in children with ALL. The decrease in serum al-bumin level may indicate the occurrence of malnutrition. Dynamic monitoring of nutritional status in children with ALL is necessary.
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Objective To investigate the effects of Danusertib on the changes in Aurora kinase B (Aurora B)/ribosomal protein p70S6 kinase (p70S6K)/ribosomal protein 15 (RPL15) signaling pathway and autophagy in human leukemia cells and its mechanism.Methods Myeloid leukemia cell lines THP-1 and K562 were selected as the research subjects.The experiment was divided into 2 phases.Phase 1:each cell line was treated with the concentration of Danusertib in 0.1 p mol/L,1.0 p mol/L and 5.0 μ mol/L.In control group,2 mL/L dimethyl sulfoxide (DMSO)was given.All the treated cells were cultured for 24 hours.The viability of each cell line was examined by methyhhiazoletrazolium assay and the autophagy was assessed by flow cytometry.In addition,the protein levels of p70S6K,AuroraB,phosphatidylinositol 3-kinase (PI3K),AKT(phosphorylated protein kinase B),mammalian target of rapamycin (mTOR),microtubule-associated protein (LC3),Beclin1 and RPL15 were determined by using Western blot.Part 2:Aurora B and RPL15 were down-regulated in THP-1 and K562 cells,respectively.DMSO was used to dissolve Danusertib(5.0 μ mol/L).The grouping was designed as following:DMSO group (blank control group),Danusertib treated group,empty plasmid group,small interfering RNA(siRNA) group,empty plasmid + Danusertib-treated group and siRNA + Danusertib treated group.The protein levels of Aurora B,p70S6K,RPL15,Beclinl and LC3 were detected by using Western blot.Results (1) Danusertib decreased the viability of THP-1 and K562 cells and the half maximal inhibitory concentration values were 26.9 pmol/L and 30.2 μmol/L for THP-1 and K562 cells,respectively.(2)The protein levels of p-Aurora B/Aurora B,p-p70S6K/p70S6K,RPL15,p-mTOR/mTOR and p-AKT/AKT decreased compared with control cells after being treated with 0.1 μmol/L,1.0 μ mol/L and 5.0 pmol/L of Danusertib in THP-1 and K562 cells,and the differences were statistically significant (all P ＜ 0.05).(3) In THP-1 cells,compared with the empty plasmid group,the protein levels of p70S6K and RPL15 decreased by 22.1％,61.3％ (F =18.1,P =0.001) and 55.4％,56.1％ (F =19.4,P =0.001) in siRNA group and siRNA + Danusertib-treated group after knockdown of Aurora B.In contrast,the protein levels of LC3 increased by 13.6％ and 17.1％ (F =15.4,P =0.001)compared with the empty plasmid group.In addition,the protein levels of Beclin1 and LC3 increased by 39.5％,92.3％ (F=25.2,P=0.001) and 40.2％,58.3％ (F=23.9,P=0.001) in siRNA group and siRNA + Danusertib treated group,compared with the empty plasmid group after down-regulation of RPL15.In K562 cells,compared with the empty plasmid group,the protein levels of p70S6K and RPL15 decreased by 24.2％,62.7％ (F =20.4,P=0.001) and 57.2％,60.1％ (F =23.9,P =0.001) in siRNA group and siRNA + Danusertib treated group after downregulation of Aurora B.But the protein levels of LC3 increased by 17.9％ and 56.7％ (F =20.9,P =0.001)compared with the empty plasmid group.Moreover,the protein levels of Beclin1 and LC3 were increased by 20.6％,98.4％ (F=22.4,P =0.001) and 41.5％,70.1％ (F=26.2,P =0.001) in siRNA group and siRNA + Danusertib treated group,compared with the empty plasmid group after downregulation of RPL15.Conclusion Danusertib can induce autophagy via inhibition of the PI3K/AKT/mTOR signaling pathway and can negatively regulate Aurora B/p70S6K/RPL15 axis in THP-1 and K562 cells.In addition,RPL15 may be a key target of Aurora B/p70S6K/RPL15signaling pathway in the inhibition of tumor proliferation.
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OBJECTIVE@#: To observe the expression of gene in the early development stage of wild zebrafish embryos.@*METHODS@#: The collinearity of gene and the sequence similarity of G6pd protein were analyzed with gene database and BLAST software, respectively. Expression of gene in different development stages of zebrafish embryos was detected by hybridization. The -EGFP-pCS recombinant plasmids were microinjected into zebrafish embryos, and fluorescence was observed under a fluorescence microscope. The expression of G6pd protein at 24, 48 and 72 hour post fertilization (hpf) zebrafish embryos was detected by Western blotting; the enzyme activity of G6pd at 24, 48 and 72 hpf zebrafish embryos was detected by modified G6pd quantitative ratio method.@*RESULTS@#: The G6pd protein similarity of zebrafish and human was 88%, and that of zebrafish and mouse was 87%. The results of hybridization showed that the gene was mainly expressed in the hematopoietic tissues of zebrafish; the results observed after microinjection of -EGFP-pCS recombinant plasmid were consistent with the results of hybridization. At 24, 48 and 72 hpf, the relative expression levels of G6pd protein in zebrafish embryos were 1.44±0.03, 1.47±0.05, and 1.54±0.02, respectively(>0.05); the G6pd enzyme activity levels were 1.74±0.17, 1.75±0.12, 1.71±0.22, respectively (>0.05).@*CONCLUSIONS@#: The study has observed the expression of gene and G6pd protein, and G6pd enzyme activity in zebrafish embryos at different development phases, which provides a reference for the establishment of a zebrafish G6PD deficiency model.
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Animals , Humans , Mice , Embryo, Nonmammalian , Gene Expression Regulation, Developmental , Glucosephosphate Dehydrogenase , Genetics , In Situ Hybridization , Plasmids , Genetics , Zebrafish , Embryology , GeneticsABSTRACT
Objective To retrospectively analyze the effect of three-dimensional radiotherapy on the survival of patients with stage Ⅳ squamous cell lung cancer.Methods Of the 101 patients collected from two prospective phase Ⅱ studies, 88 were part of the per-protocol set.All patients received platinum-doublet chemotherapy with concurrent radiation to the primary tumor.Primary endpoints were overall survival (OS) and progression-free survival (PFS).Survival was calculated using the Kaplan-Meier estimator, and univariate and multivariate analyses were performed using the log-rank test and Cox model, respectively.Results The 1-, 2-, 3-, and 5-year OS rates of the 88 patients were 42.2%, 13.6%, 8.7%, and 3.1%, respectively, and the median survival time (MST) was 10 months.The 1-, 2-, 3-, and 5-year OS and MST at PTV dose ≥63 Gy were 45.7%, 25.7%, 17.1%, 7.1%, and 11 months, respectively, whereas the 1-, 2-, 3-, and 5-year OS and MST at PTV dose<63 Gy were 39.6%, 4.5%, 2.8%, 0%, and 10 months, respectively (P=0.007).The median PFS at ≥63 Gy and<63 Gy were 9 months and 7 months, respectively (P=0.032).The 1-, 2-, 3-, and 5-year OS and PFS of patients who received 4 cycles of chemotherapy at a PTV dose of ≥63 Gy were 51.9%, 29.6%, 18.5%, 9.9%, and 9 months, respectively (P=0.001 and P=0.012), which were significantly prolonged compared with other treatment modalities.Multivariate analysis showed that PTV ≥63 Gy may be influence the OS of patients (P=0.080).Conclusions Three-dimensional radiotherapy can prolong the survival of patients with stage ⅠV squamous cell lung cancer, as demonstrated by the gradual improvement in OS and PFS following the increase in the intensity of concurrent chemotherapy and radiation therapy.A PTV dose of ≥63 Gy may be influence the OS.
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Objective To explore the effects of vinblastine on abcb4 tumor resistance gene expression in early development zebrafish and lay an important foundation for multi-drug resistant antineoplastic screening in zebrafish model.Methods Zebrafish embryos of 0.5~1.5 hours post-fertilization were exposured to different concentrations of vinblastine, and then calculated the IC50 of vinblastine.Zebrafish embryos of 0.5~1.5 hours post-fertilization were treated with different concentrations of vinblastine and embryo culture medium respectively, and then observed zebrafish embryo development in 24~120 hours post-fertilization and recorded the number of death, hatch and malformation.Evaluating the impact of vinblastine on abcb4 gene expression in zebrafish with quantitative real-time PCR and whole-mount in situ hybridization by collecting zebrafish embryos exposed to different concentrationsof vinblastine.Results Vinblastine IC50 in zebrafish embryos was 3.08 μmol/L.The mRNA level of abcb4 gene in vinblastine treated embryos was significantly increased compared to blank control group.Moreover, abcb4 gene positive hybridization signals were found in the small intestine of zebrafish embryos after 120 hours post-fertilization and also found in the brain and heart of zebrafish embryos by the method of whole-mount in situ hybridization.Conclusions Vinblastine can significantly increase the expression level of abcb4 tumor resistancegene in early development zebrafish embryos, which indicates that zebrafish can be used as a tumor resistant drug screening model.
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BACKGROUND: Mesenchymal stem cells attract extensive attention because of good biological characteristics and broad prospects, but the cells gradually show the characteristics of the aging with the increase of individual age or incubation time in vitro. Nonhuman primates have similar biological characteristics with human being, and have unique advantage in the animal model and disease treatment research.OBJECTIVE: To analyze the difference in proliferation and differentiation of bone marrow mesenchymal stem cells from macaques at different ages and to explore the effect of age on bone marrow mesenchymal stem cells and the possible mechanism.METHODS: Bone marrow samples from male macaques aged < 3 years and over 20 years were collected through bone marrow puncture, and divided into young group and elder group, with three macaques in each group. Then, bone marrow mesenchymal stem cells were isolated and cultured in vitro, and the morphological changes, proliferation and differentiation ability were observed. Age-related beta-galactosidase staining was performed, and protein microarray and ELISA were used to detect cytokine levels.RESULTS AND CONCLUSION: With age, the proliferation and differentiation of bone marrow mesenchymal stem cells from the elder macaques were reduced significantly, and the number of senescent cells increased significantly; the levels of interleukin-1b, interleukin-4, interleukin-6, tumor necrosis factor α and vascular endothelial growth factor were elevated obviously, the levels of heparin-binding basic fibroblast growth factor and placental growth factor were reduced. These findings indicate that the body's aging lead to the reduction in the proliferation, differentiation and cytokine secretion of bone marrow mesenchymal stem cells.
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Objective To illustrate that tumor necrosis factor-α (TNF-αt) / nuclear factor of activated T cells pathway is the main function way that mesenchymal stem cells (MSC) inhibit proliferation of pulmonary vascular smooth muscle cells during treating pulmonary hypertension (PH).Methods MSC from human umbilical cord was used to treat PH rat models induced by monocrotaline.Rats were divided into the control group,the PH model group and the MSC group.The general conditions of the rats were observed.Haemodynamics was detected.Pathological sections and immunohistochemistry method were used to detect the lung structure and tissue changes.Changing conditions of TNF-αt/NFAT were detected.Results Compared with rats in the PH model group,the general conditions of the MSC group tended to be normal evidently:the right ventricular systolic pressure (RVSP) dropped [(30.37 ±3.13) mmHg vs.(47.90 ± 3.45) mmHg,1 mmHg =0.133 kPa],the aortic pressure (MAoP) increased [(115.03 ± 16.01) mmHg vs.(92.78 ± 16.28 mmHg)],the thickening condition of arterial intima-media was evidently relieved [(17.22 ±1.21)％ vs.(31.68 ±2.26)％],the plasma TNF-α level decreased obviously [(842 ±76) ng/L vs.(245 ±24)ng/L],and the lung tissue TNF-o level decreased (0.172 ±0.024 vs.0.248 ± 0.051),and all the differences were statistically significant (all P ＜ 0.05).The activation of pulmonary artery NFATc2 in the MSC treatment group was apparently inhibited.Conclusions MSC therapy may perform the treating effect in PH by inhibiting the over-proliferation of inflammation related pulmonary vascular smooth muscle cells via TNF-oα/NFAT pathway.
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Thalassemia is inherited hemolytic anemia caused by globin production disorder,clinically known as beta thalassemia and alpha thalassemia resulted from beta and alpha chain synthesis disorder.Significant anemic clinical manifestations are observed in intermediate -level and severe -level patients with the 2 types of anemia, affecting greatly the growth and life quality of children patients who need blood transfusion to stay alive.The incidence rate of thalassemia is as high as 7% -20% in South China,so the prevention and standaridized treatment of thalasse-mia and the management over target population draw more attention.This paper is a review on the latest development of blood transfusion,chelation therapy,health education,birth intervention for thalassemia.
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Objective To investigate the effects of gefitinib on early embryonic development and expression of abcb4 tumor resistance genes in zebrafish.Methods Zebrafish were adopted as experimental animals and divided into gefitinib group,mixture of doxorubicin and gefitinib group and blank group.Zebrafish embryos of 0.5-1.5 hours after fertilization(0.5-1.5 hpf) were exposed to different concentrations of gefitinib,and then embryo development of zebrafish in 24-120 hpf was observed and the number of death,hatch and malformation was recorded.The embryo mortality was calculated under different concentrations of gefitinib at different time points,and the numerical value of IC50 was calculated;Hatching rates of zebrafish embryo in 48 hpf and 72 hpf and malformation rates of zebrafish embryo in 120 hpf were calculated.The zebrafish embryos exposed to different concentrations of gefitinib in different groups were collected,and the expressionof abcb4 gene in zebrafish embryos was detected by real-time quantitative PCR.Results Gefitinib IC50 in zebrafish embryos was 16.18 μmol/L.Compared with the control group,higher dosage (20 μmol/L) of gefitinib in other two groups significantly decreased hatching rates of embryos,and had obvious embryonic lethal effects and teratogenic effects.Moreover,the mRNA levels of the abcb4 gene in the zebrafish embryos of gefitinib group were not significantly changed,whereas the mRNA levels of the abcb4 gene in mixture of doxorubicin and gefitinib group were significantly different(P＜0.05).Conclusion Gefitinib has no significant effects on the expression of abcb4 tumor resistance gene in early development of zebrafish embryos(P＞0.05),but it can reverse the drug resistant effects of doxorubicin,suggesting that zebrafish can construct tumor resistance model.