紫杉醇联合顺铂化疗对口腔癌患者外周血细胞免疫的影响

目的分析紫杉醇联合顺铂化疗对口腔癌患者外周血细胞免疫等指标的影响。方法根据化疗方案不同,将武警广东总队医院于2019年2月至2020年1月收治的100例口腔癌患者分为对照组50例(紫杉醇化疗)、观察组50例(紫杉醇联合顺铂化疗)。对比2组治疗前后的外周血细胞免疫指标、治疗效果、化疗不良反应以及生活质量变化。结果①外周血细胞免疫指标:治疗前各项指标相比差异无统计学意义(P>0.05),治疗后,观察组患者CD4^(+)/CD8^(+)、CD4^(+)、CD3^(+)水平高于对照组、CD8^(+)水平低于对照组(P<0.05);②治疗效果:观察组治疗有效率84%高于对照组(P<0.05);③化疗不良反应:对照组不良反应发生率为22%,观察组为20%,差异无统计学意义(P>0.05);④生活质量:观察组生活质量改善率高于对照组(44%与20%,P<0.05)。结论在口腔癌化疗中联合应用紫杉醇和顺铂可以有效调节细胞免疫功能,改善生活质量,且安全性有保证。

不同辐照剂量率X射线对小鼠外周血细胞及免疫器官的影响

目的探讨不同辐照剂量率X射线对小鼠外周血细胞及免疫器官的影响。方法 120只小鼠随机分为正常对照组、低剂量率照射组和高剂量率照射组,每组40只,雌雄各半。分别于照射后24h、48h、96h、192h每组各取10只小鼠摘眼球取血用于测定红细胞计数(RBC)、白细胞计数(WBC)、血小板数(PLT)和血红蛋白(HGB)含量,然后处死小鼠取胸腺和脾脏称重,计算胸腺指数和脾脏指数。另取一部分做组织匀浆,测定匀浆中的超氧化物歧化酶(SOD)和丙二醛(MDA)含量。结果不同剂量率X射线照射均会造成小鼠外周血细胞中RBC、WBC、PLT和HGB的降低,高、低剂量率组间无显著差异。在相同取样时间,高、低剂量率组小鼠胸腺、脾脏指数与对照组比较均差异显著,高、低剂量率组间脾脏指数差异显著,在取样时间24 h,高、低剂量率组间胸腺指数差异显著。高、低剂量率组小鼠胸腺、脾脏组织匀浆中MDA的含量均升高,SOD含量均降低,且与对照组比较差异显著。高、低剂量率组小鼠胸腺组织匀浆中MDA的含量在取样时间48 h、96 h差异显著,SOD的含量在取样时间24 h、48 h、96 h差异显著;脾脏组织匀浆中MDA的含量在取样时间24 h、48h差异显著,SOD的含量在取样时间24 h、48 h、192 h差异显著。结论高、低剂量率辐照对小鼠外周血细胞数量的影响无显著差异;高剂量率辐照对小鼠脾脏指数的影响较低剂量率大,对胸腺指数的影响无显著差异;高剂量率辐照小鼠胸腺、脾脏组织中SOD和MDA的影响较低剂量率大。

Effect of in vitro interferon-beta administration on hepatitis C virus in peripheral blood mononuclear cells as a predictive marker of clinical response to interferon treatment for chronic hepatitis C

AIM:To test whether in vitro incubation of peripheral blood mononuclear cells (PBMC) with interferon (IFN) could efficiently decrease hepatitis C virus-RNA (HCV-RNA) amount and to analyze whether this effect was associated with clinical response to IFN.METHODS:Twenty-seven patients with histologically proven chronic hepatitis C were given intravenous administration of 6 million units (MU) IFN-β daily for 6 weeks followed by three times weekly for 20 weeks. PBMC collected before IFN therapy were incubated with IFN-β and HCV-RNA in PMBC was semi-quantitatively determined.RESULTS: Twenty-five patients completed IFN therapy.Eight patients (32%) had sustained loss of serum HCV-RNA with normal serum ALT levels after IFN therapy (complete responders).HCV-RNA in PBMC was detected in all patients,whereas it was not detected in PBMC from healthy subjects.In vitro administration of IFN-β decreased the amount of HCV-RNA in PMBC in 18 patients (72%). Eight of these patients obtained complete response. On the other hand,none of the patients whose HCV-RNA in PBMC did not decrease by IFN-β was complete responders. Multiple logistic regression analysis revealed that the decrease of HCV-RNA amount in PBMC by IFN-β was the only independent predictor for complete response (P<0.05).CONCLUSION:The effect of in vitro IFN-β on HCV in PBMC reflects clinical response and would be taken into account as a predictive marker of IFN therapy for chronic hepatitis C.

Decreased mitochondrial deoxyribonucleic acid and increased oxidative damage in chronic hepatitis C

AIM： To determine whether alteration of the mito- chondria DNA （mtDNA） copy number and its oxidative damage index （mtDNA△CT） can be detected by analysis of peripheral blood cells in hepatitis C virus （HCV）- infected patients. METHODS： This study enrolled two groups of pa- tients aged 40-60 years： a control group and an HCV- infected group in Department of Gastroenterology and Hepatology in Changhua Christian Hospital. Patients with co-infection with hepatitis B virus or human im- munodeficiency virus, autoimmune disease, malignant neoplasia, pregnancy, thyroid disease, or alcohol con- sumption 〉 40 g/d were excluded. HCV-infected pa- tients who met the following criteria were included： （1） positive HCV antibodies for 〉 6 mo; （2） alanine aminotransferase （ALT） levels more than twice the upper lim- it of normal on at least two occasions during the past 6 mo; and （3） histological fibrosis stage higher than F1. The mtDNA copy number and oxidative damage index of HCV mtDNA （mtDNA△CT） were measured in periph- eral blood leukocytes. The association between mtDNA copy number and mtDNA△CT was further analyzed using clinical data. RESULTS： Forty-seven normal controls （male/female： 26/21, mean age 50.51 ± 6.15 years） and 132 HCV- infected patients （male/female： 76/61, mean age 51.65 ± 5.50 years） were included in the study. The geno- types of HCV-infected patients include type 1a （n = 3）, type 1b （n = 83）, type 2a （n = 32）, and type 2b （n = 14）. Liver fibrosis stages were distributed as follows： F1/F2/F3/F4 = 1/61/45/25 and activity scores were A0/ A1/A2/A3 = 7/45/55/25. There were no age or gender differences between the two groups. HCV-infected pa- tients had higher hepatitis activity （aspartate transami- nase levels 108.77 ± 60.73 vs 23.19 ± 5.47, P 〈 0.01; ALT levels 168.69 ± 93.12 vs 23.15 ± 9.45, P 〈 0.01） and lower platelet count （170.40±58.00 vs 251.24 ± 63.42, P 〈 0.01） than controls. The mtDNA copy num- ber was lower in HCV-infected patients than in controls （173.49 vs 247.93, P 〈 0.05）. The mtDNA△CT was higher in HCV-infected patients than in controls （2.92 vs 0.64, P 〈 0.05）. To clarify the clinical significance of these results in HCV-infected patients, their association with different clinical parameters among HCV-infected pa- tients was analyzed. A negative association was found between mtDNA copy number and elevated aspartate transaminase levels （r = -0.17, P 〈 0.05）. Changes in mtDNA copy number were not associated with HCV RNA levels, HCV genotypes, liver fibrosis severity, or inflammatory activity in the liver biopsy specimen. How- ever, a correlation was observed between mtDNA△CT and platelet count （r = -0.22, P 〈 0.01）, HCV RNA level （r = 0.36, P 〈 0.01）, and hepatitis activity （r = 0.20, P = 0.02）. However, no difference in the change in mtDNA△CT was observed between different fibrosis stages or HCV CONCLUSION： Oxidative stress and mtDNA dam- age are detectable in patient＇s peripheral leukocytes. Increased leukocyte mtDNA△CT correlates with higher HCV viremia, increased hepatitis activity, and lower platelet count.

Inflammatory pseudotumor of the liver and peripheral eosinophilia in autoimmune pancreatitis

AIM: Inflammatory pseudotumor (IPT) of the liver is a rare benign lesion, the etiology of which remains obscure.It is not associated with any particular diseases apart from phlebitis and Crohn's disease.METHODS: A middle-aged male with hepatic IPT and peripheral eosinophilia associated with autoimmune pancreatitis (AIP) was selected for this study and review of literature.RESULTS: A 59-year-old male was admitted with obstructive jaundice, marked eosinophilia (1 343/mm3)and hypergammaglobulinemia (4 145 mg/dL). Imaging techniques revealed dilatation of the intrahepatic bile duct,stenosis of the common bile duct with diffuse wall thickening, gallbladder wall thickening, irregular narrowing of the pancreatic duct, and swelling of the pancreatic parenchyma. Multiple liver masses were also demonstrated and diagnosed as IPT by biopsy specimens. Six months later, the abnormal features of the biliary tree remarkably improved by the oral administration of prednisolone, and the liver masses disappeared. The swelling of the pancreatic head also improved. The peripheral eosinophil count normalized. IPT associated with AIP, as we know,has not been reported in the literature. The clinical features of the present case mimicked those of pancreatic cancer with liver metastasis. This case deserves to be documented to prevent misdiagnosis of similar cases.

Detection of Intracellular Cytokine Production in Peripheral Blood CD_^+ T Cells of Patients with Recurrent Genital Herpes

Objective: To study the role of Th1/Th2 cytokines in thepathogenesis of recurrent genital herpes (RGH), and tobetter understand the relationship between them. Methods: A two-color immunofluorescent staining of cellsurface antigen and intracellular cytokines for flowcytometric analysis was used for CD_3, IL-2, IL-10, IL-12,IFN-γ and TNF-α in CD_3^+ T-lymphocytes in activatedperipheral blood mononuclear cells of patients with RGH. Results: Compared to controls, patients with RGHshowed fewer CD_3^+ T cells (P<0.05) and IL-2 producing andIFN-γ producing T cells (P<0.02 and P<0.001, respectively)after in vitro stimulation with PMA and ionomycin in thepresence of a protein transport inhibitor. More IL-10producing and IL-12 producing T cells were found inpatients with RGH (P<0.01). There was no significantdifference in the number of TNF-α producing cells betweenRGH patients and controls (P<0.05). Conclusion: RGH patients showed relatively more Th2cytokines. The imbalance between Th1 and Th2 cytokinesresults in inhibitory effects on a series of cell-immuneresponses, which may play an important role in thepathogenesis of RGH.

Mechanism of peripheral blood mononuclear cell invasion by HBV on artificial immunization in newborns

OBJECTIVE: To study the effect and mechanism of the peripheral blood mononuclear cell (PBMC) invasion by HBV on artificial immunization in newborns. METHODS: Fifty-two newborns of HBsAg positive mothers were immunized with HBIG (hepatitis B immunoglobulin) and HBVac (hepatitis B vaccine) and were followed up for 7 months. The newborns' HBV-DNA in serum and in the PBMCs was detected with nested-PCR; anti-HBs was tested with solid phase radioimmunoassay (SP-RIA). PBMCs isolated from newborn peripheral blood were incubated in the presence of PHA or purified HBsAg. Interleukin-2 (IL-2) level in culture supernatants of activated cells was detected by ELISA. RESULTS: The failure rate of immunization was higher in infants with positive HBV-DNA in PBMCs than those with negative HBV-DNA (P

A simple method for removing low-density granulocytes to purify T lymphocytes from peripheral blood mononuclear cells

Objective： Low-density granulocytes （LDGs） can form neutrophil extracellular traps （NETs） spontaneously and excessively. When peripheral blood mononuclear calls （PBMCs） are used for studying T lymphocytes, LDGs contained in the PBMCs may decrease the threshold of activating T lymphocytes by forming NETs. This study focused on the profiles of LDGs in common autoimmune diseases and methods for removing LDGs from PBMCs. Methods： The percentages of LDGs in PBMCs from 55 patients with dermatomyositis （DM）, 15 with polymyositis （PM）, 42 with rheumatoid arthritis （RA）, 25 with systemic lupus erythematosus （SLE）, and 19 healthy controls were determined by flow cytometry. Three methods of removing LDGs were explored and compared. After removal, PBMCs from six patients with positive T-SPOT.TB were tested again to find out if LDGs contained in the PBMCs could influence T lymphocyte reactions. Results： Significantly higher LDG percentages were found in PBMCs from patients with DM （（8.41±10.87）%, P〈0.0001 ）, PM （（8.41±10.39）%, P〈0.0001 ）, RA （（4.05±6.97）%, P=0.0249）, and SLE （（7.53±11.52）%, P=0.0006）, compared with the controls （（1.28±0.73）%）. The T-SPOT.TB values significantly decreased after LDGs were removed. Increasing relative centrifugal force （RCF） within a limited range can decrease the LDG percentage from an initial high level, but not markedly increase the LDG clearance rate. Compared with the whole blood sediment method, the PBMC adherence method can significantly remove LDGs yet scarcely influence the T lymphocyte percentage in PBMCs. Conclusions： The LDG percentage in PBMCs is significantly increased in patients with S/E, DM, PM, and RA. The influence of LDGs on T lymphocytes cannot be ignored in PBMC cultures. The adherence method is a simple and easy-to-use method for removing LDGs and purifying T lymphocytes from PBMCs.