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Objective:To investigate the difference in the radiation sensitivity of hematopoietic stem and progenitor cells (HSPCs) derived from fetal liver and bone marrow.Methods:HSPCs from fetal liver of 14.5 d embryo or bone marrow of 8 week-old mice were isolated to receive a single dose of 5 or 10 Gy irradiation in vitro using a 60Co irradiator. Twelve hours later, the cell apoptosis, mitochondrial reactive oxygen species (ROS) level, colony formation ability and DNA damage in HSPCs were detected. Freshly isolated HSPCs were injected into lethally irradiated CD45.1 + C57BL/6J mice (4.5 Gy+ 5 Gy with an interval of 30 min) Chimerism rate, lineage constitution, and cell cycle were analyzed 12 weeks after transplantation. Results:Compared with bone marrow HSPCs after irradiation, the percentage of apoptosis in fetal liver HSPCs was significantly higher ( t=16.21, 12.27, P<0.05), the level of ROS was dramatically elevated ( t=68.72, 18.89, P<0.05). At 10 Gy, fetal liver HSPCs could not form colonies at all ( t=12.41, 15.67, 9.46, P<0.05). γ-H2AX immunofluorescence staining showed that the DNA damage of fetal liver HSPCs was more severe after irradiation, and the number of Foci formed was significantly higher than that of bone marrow HSPCs ( t=2.27, 2.03, P< 0.05), which indicated that fetal liver HSPCs were more sensitive to radiation. The chimerism rate of transplanted fetal liver HSPCs was lower than that of bone marrow cells ( t=5.84, P<0.05) with a higher proportion of myeloid lineage, suggesting that fetal liver HSPCs had lower in vivo reconstitution capacity than bone marrow HSPCs and were more prone to myeloid differentiation. The cell cycle of bone marrow HSPCs from transplanted chimeric mice was examined, and the proportion of S-phase was significantly higher in the fetal liver group than that in the bone marrow group ( t=2.89, P<0.05). Mitochondrial stress results showed that fetal liver HSPCs had higher basal respiratory capacity ( t=39.19, P<0.05), proton leakage ( t=6.64, P<0.05), ATP production ( t=9.33, P<0.05), and coupling efficiency ( t=7.10, P<0.05) than bone marrow c-Kit + cells, while respiratory reserve capacity ( t=5.53, P< 0.05) was lower than that of bone marrow c-Kit + cells. Conclusions:HSPCs derived from fetal liver display higher radiosensitivty compared with bone marrow HSPCs, laying the foundation for an in-depth illustration of the effects of radiation on hematopoietic stem cells at different developmental stages.
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Objective:To systematically evaluate the clinical efficacy of compound α-ketoacid tablets in the treatment of diabetic kidney disease (DKD).Methods:CNKI, Wanfang database, EMBASE, PubMed and Cochrane Library database were searched for eligible records published from the establishment of individual database to November 13 th, 2022. The quality of the included studies were assessed, data were extracted, and meta-analysis was conducted using RevMan5.3. Results:A total of 26 randomized controlled trials were included, with a total of 2 790 DKD patients (1 465 in the experimental group and 1 325 in the control group). Multiple parameters were significantly improved in the experimental group compared with the control group, including 24-hour urinary protein, blood creatinine, urea nitrogen, nutritional index, oxidative stress level, fasting blood glucose, glycated hemoglobin, homocysteine, HGF, VEGF, TGF-β1, and systolic blood pressure.Conclusions:Limited low-quality evidence showed that compound α-ketoacid tablets combined with low-protein diet may be related to the improved 24-hour urinary protein, renal function, and glucose metabolism in patients with DKD. Due to the lack of randomized controlled trials designed for respective stages of DKD, the inclusion criteria of our study were relatively general, possibly leading to the lack of pertinence of the results. Some indicators showed apparent heterogeneity among different groups, and more high-quality multi-center studies with large sample sizes are still needed to verify our findings.
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Objective:To investigate the effect and underlying mechanism of BRCC3 knockout on acute GVHD(aGVHD) of mice.Methods:A total of 12 recipient C57BL/6J mice were divided into two groups, including 6 wild type(WT) and BRCC3 -/-(KO). The recipients were exposed to 4.5 Gy + 4.5 Gy 60Co γ-rays in total body irradiation (TBI) at 30 min intervals. At 6 h post-irradiation, 1×10 7bone marrow cells and 8×10 6 splenocytes from BALB/c mice were infused into C57BL/6J mouse via tail vein to develop aGVHD mouse model. BRCC3 was specifically knocked out in aGVHD mouse model. The organ damage was examined through histopathology. The levels of serum cytokines were measured by enzyme-linked immuno sorbent assay (ELISA) and cytometric bead array (CBA), respectively. Spleen, liver and small intestine lymphocytes were isolated at 9 d post-transplantation, and the infiltration and activation of T cells in the target organs were assayed using flow cytometry. Results:The absence of BRCC3 in recipient mice significantly shortened survival ( P<0.05) with increased liver injury of aGVHD mice. In BRCC3 -/-recipient mice, the proportions of CD8+ T cells and CD8+ CD25+ T cells were significantly higher than those in the spleen( t=6.53, 5.52, P<0.05), and the proportions of CD8+ T cells and CD8+ CD25+ T cells were significantly increased in the liver ( t=3.74, 3.19, P<0.05). Similarly, the proportions of CD8+ T cells, CD8+ CD25+ T cells and CD8+ CD69+ T cells were significantly elevated in the small intestine ( t=3.52, 4.06, 3.29, P<0.05). Conclusions:BRCC3 deletion increased the proliferation and activation of donor CD8+ T cells and aggravated aGVHD, which might provide a new prevention and treatment target for aGVHD.
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Objective:To study the effect of different doses of 60Co γ-ray ionizing radiation on mitochondrial function in mouse hematopoietic stem and progenitor cells (HSPCs). Methods:C57BL/6 mice were divided into control group, 1 Gy irradiation group and 4.5 Gy irradiation group. The mitochondrial functions were detected at 12 h and 24 h after irradiation, including ROS level, membrane potential, mitochondrial structure, and mitochondrial stress. Bone marrow c-Kit + cells received a single 15 Gy irradiation in vitro, after 24 h, mitochondrial function was detected. Results:It was found that mice leukocytes ( t=12.41, 18.31, 16.48, 14.16, 19.08, 20.25, P<0.05), red blood cells ( t=4.81, 6.62, P<0.05) and platelets ( t=4.33, 6.68, P<0.05) were significantly reduced. The numbers of bone marrow colony formation unit ( t=16.27, 55.66, 17.06, 43.75, P<0.05), and HSPCs ( t=5.16, 11.55, P<0.05) were decreased dose-dependently post-irradiation. Under 1 Gy irradiation, the mitochondrial function and mitochondrial basal metabolic index of HSPCs ( t= 7.36, 3.68, 4.58, 3.15, 3.15, P<0.05) were enhanced at 24 h post-irradiation. Under 4.5 Gy irradiation, mitochondrial number, mitochondrial membrane potential ( t=12.29, 10.46, P<0.05), maximal respiration and spare respiratory capacity were decreased ( t=7.81, 5.78, 6.70, 5.83, P<0.05), ROS level was increased ( t=4.63, 4.12, P<0.05). The basal respiration and oxidative phosphorylated ATP production were reduced at 12 h after irradiation ( t=8.48, 3.80, P<0.05); and the proton leakage was increased ( t=6.57, P<0.05) and coupling efficiency was reduced ( t=11.43, P<0.05) at 24 h after irradiation. In cultured c-Kit + cells, the level of ROS ( t=11.30, P<0.05) and the maximum respiration and spare respiratory capacity were increased ( t=4.25, 3.44, P<0.05) while the mitochondrial membrane potential was decreased ( t=34.92, P<0.05) significantly. Conclusions:A method for systematically assessing mitochondrial function in HSPCs was established, and the effect of ionizing radiation on mitochondrial function of HSPCs was clarified, laying a foundation for further revealing the mechanism of ionizing radiation-induced mitochondrial damage in HSPCs.
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Objective: To investigate the entry points of traditional Chinese clinical pharmacists participating in multidisciplinary team (MDT) to provide reference for more peers and subject development. Methods: According to the daily work of traditional Chi-nese clinical pharmacists in joint multidisciplinary clinic and combining with patients' medication characteristics, traditional Chinese clinical pharmacists carried out works from three aspects including doctors, patients and peers. Results: After the medication was guided by traditional Chinese clinical pharmacists, the effect of medicinal treatment was improved, the patients' medication compliance increased and the incidence of adverse reactions was reduced. Moreover, it brought more ideas to the peers within the discipline in our hospital. Conclusion: Traditional Chinese clinical pharmacists can exploit their advantages to the full to find clinical demands and more entry points in works. At the same time, traditional Chinese clinical pharmacists should enhance related knowledge, professional qualities, and cooperation and communication ability comprehensively.
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Objective To investigate the effects of four tyrosine metabolites on the proliferation, cell cycle and chemosensitivity of lung cancer cells.Methods A549 cells were cultured with different concentrations of tyrosine metabolites:tyrosine, 4-hydroxyphenylpyruvate(HPPA), homogentisic acid(HGA) and fumaric acid (FA) in vitro.High content imaging analysis system was used to monitor cell proliferation.Flow cytometry was used to detect cell cycle and apoptosis.Protein and mRNA level of Bax and Bcl-2 were measured by Western blot.Results Tyrosine metabolites had no effect on cell cycle and proliferation of A549 cells.HGA decreased the cell apoptosis of A549 cells induced by Gefitinib with depressed Bax expression and elevated Bcl-2 expression.However, FA increased the apoptosis of A549 cells induced by Gefitinib with up-regulation of Bax and down-regulation of Bcl-2.No obvious effect of tyrosine and HPPA was detected on Gefitinib induced cell apoptosis.Conclusion HGA and FA may play an important role in drug sensitivity of lung cancer cells, indicating a critical role of tyrosine metabolism in lung cancer.
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Objective To introduce the working experience of clinical pharmacists who provide pharmaceutical service in participating integrated diabetes out-patient.Methods Review and analyze the patients' clinical features and the clinical pharmacists' working model and effect in integrated diabetes out-patient of the Sixth People's Hospital Affiliated to Shanghai Jiaotong University.Results Among these patients who got pharmaceutical service in integrated diabetes out-patient, 81.7% were 40 to 70 years old, 51.7% had diabetes for 5 to 10 years, 50.0% were taking oral antidiabetic drugs to control blood glucose, and 90.0% were having complications.After the medication guide by clinical pharmacists,patients' medication compliance was increased from the percentage of 30.0% to 80.0%, the good control rate of blood glucose was increased from the percentage of 8.3% to 41.7%, and the incidence rate of adverse reaction was reduced from 10.0% to 3.3%.Conclusion Clinical pharmacists who actively participate in integrated diabetes out-patient could ensure the effective and safe treatment to diabetic patients.Therefore, this model is worth to be popularized due to professional value of clinical pharmacists.
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Objective To generate the erythroid differentiation associated gene(EDAG) knockout mice and analyze their sensitivity to low dose radiation-induced damage.Methods Zinc finger nuclease technology ( ZFNs ) was used to produce the EDAG knockout mice.The low dose radiation-induced damage was evaluated by peripheral blood cell counts, DNA damage and colony formation of bone marrow cells.Wild-type and EDAG knockout mice were irradiated with 0.31 Gy/min X-ray, one minute per day for seven consecutive days, and the cumulative radiation dose was 2.17 Gy(n=7).The blood cell counts were measured by an automated hemocytometer.DNA damage was detected by immunofluorescence assay with a DNA damage marker p-H2A.x antibody (n=3).The colony formation ability of bone marrow cells was evaluated with a semi-solid culture medium(n=3).Results A model of EDAG knockout mice was established.Compared to wide type mice, white blood cell counts of EDAG knockout mice decreased significantly while the DNA damage marker p-H2A.x expression was increased on the third day after X-ray irradiation.The ability of colony-forming was reduced after 7 days of X-ray irradiation.Conclusion Our present study found that EDAG knockout mice are more sensitive to low dose radiation-induced damage as shown by decreased peripheral blood cells counts, reduced colony-forming ability of bone marrow cells, and increased DNA damage.These results suggest that EDAG knockout mice can serve as a powerful tool for evaluation of the biological effects of low-dose radiation damage.
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Antiradiation drugs, also known as radioprotective agents, can prevent humans from radiation injury,reduce the clinical symptoms of radiation sickness,promote early recovery and reduce morbidity or mortality.Early developments of such agents focused on thiol synthetic compounds, such as amifostine (WR 2721).This compound reduced mortality, but its disadvantages, such as large use and high toxicity, limited its use in clinical practice.To find a suitable radioprotective agent is crucial to reducing side effects induced by ionizing radiation and increasing survival rate in patients during radiotherapy.In this paper, the classification and mechanism of radioprotective agents are reviewed and future developments in this field are predicted.
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<p><b>OBJECTIVE</b>To investigate the association between the PNPLA3 rs738409 polymorphism and chronic hepatitis B (CH[B) in a Han Chinese population residing in Qingdao.</p><p><b>METHODS</b>Peripheral blood samples were collected from 185 CHB patients and 164 healthy controls and subjected to polymerase chain reaction (PCR) and DNA sequencing to determine the PNPLA3 genotypes. The relative risk of the rs738409 polymorphism for CHB was estimated by calculating the odds ratio (OR) and 95% confidence interval.</p><p><b>RESULTS</b>The rs738409 G allele frequency was significantly different between the CHB and control groups (31.9% vs.21.9% respectively, P less than 0.05). Compared to he rs738409 C allele, the G allele was associated with an increased risk of developing CHB (OR =1.67, 95% CI:1.18-2.34, P =0.003). Logistic regression model analysis, with adjustment for confounding factors, indicated that carriers of the PNPLA3 rs738409 GG + GC genotype had increased risk of CHB than carriers of the CC genotype (OR =1.76 ,95% CI:1.14-2.71, P =0.011).</p><p><b>CONCLUSION</b>Qingdao Han Chinese who are carriers of the rs738409 G allele are at increased risk of CHB.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Asian People , Genetics , Case-Control Studies , China , Epidemiology , Gene Frequency , Genetic Predisposition to Disease , Genotype , Hepatitis B, Chronic , Epidemiology , Genetics , Lipase , Genetics , Membrane Proteins , Genetics , Polymorphism, Single Nucleotide , Risk FactorsABSTRACT
<p><b>OBJECTIVE</b>To construct a GFP/Puro double-labeled lentiviral expression vector for CK8 silencing and assess the effects of CK8 silencing on cell apoptosis.</p><p><b>METHODS</b>The siRNA sequences of CK8 were inserted into the lentiviral expression vector GV248 and transfected into 293T cells with the packaging plasmids PMD and SPA. The lentivirus was collected at 24 and 36 h post-transfection. Flow cytometry was used to detect the virus titer and the positive cells were selected with puromycin. The knockdown of CK8 was examined by Western blotting. The effect of CK8 down-regulation on cell apoptosis induced by cisplatin was detected with Annexin V/PI staining.</p><p><b>RESULTS AND CONCLUSION</b>We successfully constructed CK8 interference lentiviral vector and obtained a stable cell line with CK8 knock-down that was sensitive to cisplatin-induced apoptosis.</p>
Subject(s)
Humans , Apoptosis , Cell Line , Down-Regulation , Genetic Vectors , Keratin-8 , Genetics , Lentivirus , Plasmids , RNA Interference , RNA, Small Interfering , TransfectionABSTRACT
Objective To investigate the efficacy of the Ivor-Lewis cervical stapled esophagogastrostomy via thorax in the treatment of middle esophageal carcinoma.Methods The clinical data of 303 patients with middle esophageal carcinoma who were admitted to the Rugao Boai Hospital (107 patients) and the Rugao People's Hospital (196 patients) from March 2005 to March 2013 were prospectively analyzed.All the patients received Ivor-Lewis stapled cervical esophagogastrectomy (Ivor-Lewis group,151 patients) or Sweet stapled cervical esophagogastrostomy (Sweet group,152 patients) according to the admission order.The intraoperative condition,perioperative complications,lymph node dissection and postoperative follow-up of the 2 groups were analyzed.All the patients were followed up via out-patient examination till December 2012.The measurement data,enumeration data and the ranked data were analyzed using the independent samples t-test,chi-square test or Fisher exact probability and Wilcoxon rank sum test,respectively.The survival curve was drawn by the Kaplan-Meier method,and the postoperative mortality rate was analyzed using the Cox proportional hazard model.Results The operation time of the Ivor-Lewis group was (239 ± 21)minutes,which was significantly longer than (188 ± 30)minutes of the Sweet group (t =11.32,P ＜ 0.05).The surgical resection rate of the Ivor-Lewis group was 98.68％ (149/151),which was significantly higher than 92.76％ (141/152) of the Sweet group (x2 =6.45,P ＜ 0.05).The positive rate of the upper resection margin of the esophagus,postoperative morbidity rate and operative were 0.67％(1/149),10.07％ (15/149) and 0 in the Ivor-Lewis group,and 0.71％ (1/141),11.35％ (16/141) and 0.71％(1/141) in the Sweet group,with no significant difference between the 2 groups (P ＞ 0.05).The number of lymph nodes dissected from the cervical-thoracic junction and the upper abdomen were 3.6 ± 1.1 and 3.5 ± 1.1 in the Ivor-Lewis group,which were significantly greater than 2.3 ± 0.8 and 2.4 ± 0.8 in the Sweet group (Z =9.96,9.02,P ＜ 0.05).The number of positive lymph nodes dissected from the cervical-thoracic junction was 0.7 ± 1.1 in the Ivor-Lewis group,which was greater than 0.3 ± 0.6 of the Sweet group,with significant difference between the 2 groups (Z =3.26,P ＜ 0.05).Of the 290 patients who received surgical treatment,273 were followed up with the follow-up rate of 94.14％ (273/290),and the median time for follow-up was 28.0 months.The 1-,2-,3-year tumor recurrence rates were 8.21％ (11/134),19.64％ (22/112) and 29.35％ (27/92) of the Ivor-Lewis group,which was significantly lower than 19.05％ (24/126),35.24％ (37/105) and 44.19％(38/86) of the Sweet group (x2=6.55,7.33,5.03,P ＜ 0.05).There were significant differences in the 1-,2-,3-year locoregional recurrence rate of the lymph nodes between the 2 groups (x2 =7.03,9.68,6.87,P ＜0.05).The 1-,2-,3-year accumulative survival rates of the Ivor-Lewis group were 90.30％ (121/134),80.36％(90/112) and 71.74％ (66/92),which were significantly higher than 80.95％ (102/126),59.05％ (62/105)and 51.16％ (44/86) of the Sweet group (x2=4.65,1 1.73,7.97,P ＜ 0.05).Conclusion Ivor-Lewis stapled cervical esophagogastrostomy via thorax has advantages of high resection rate,better safety and better quality of life of patients,and it could be an optimized design of the treatment for patients with middle esophageal cancer without intumescent lymph node of neck.