Distribution of natural killer cells and T-lymphocyte subsets in peripheral blood,gallbladder cancer and surrounding tissue

BACKGROUND: The patient with malignant tumor always show immunologic function drawback and ingravescent with tumor development, especially in the aspect of cell-mediated immunity. This study was undertaken to define the relationship between the immune function of local cells and cancer development by investigating the distribution of natural killer (NK) cells and T-lymphocyte subsets in peripheral blood, the cancer tissue and the tissue surrounding gallbladder carcinoma. METHODS: The numbers of CD4(+) and CD8(+) T-lymphocytes and NK cells were measured by flow cytometry in samples taken from gallbladder cancer tissue, the surrounding tissues and peripheral blood of 38 patients, and compared with the numbers in the peripheral blood and gallbladder tissue of 30 patients with cholecystitis as controls. RESULTS: The numbers of CD4(+) and CD8(+) T-cells and NK cells in gallbladder cancer tissues were significantly higher than those in the surrounding tissue and gallbladder with gallstone. However, the ratio of CD4(+)/CD8(+) was lower in the cancer tissue than that in the surrounding tissue and tissue from gallbladders with gallstones. The distribution of CD4(+) and CD8(+) T-cells and NK cells in mucous membrane of cholecystitis gallbladder and that in the tissue surrounding gallbladder cancer were significantly different. CONCLUSIONS: Disproportionate and imbalanced distribution of NK cells and subsets of T-lymphocytes occurs in the mucous membrane proper of gallbladder cancer and surrounding tissue. Although gallbladder cancer tissue has higher expressions of CD4(+), CD8(+) and NK cells, the immune function is low or in an inhibited state. In gallbladder cancer immunization therapy, local cellular immunological function should be enhanced and the protective barrier improved.

Effects of Intraoperative Administration of Dexmedetomidine on the Percentage of T-Lymphocyte Subsets and Natural Killer Cells in Patients with Colorectal Cancer

Study Objective: To observe the effect of dexmedetomidine (DEX) on T-lymphocyte subsets and natural killer (NK) cells in the peripheral blood of perioperative patients with colorectal cancer. Design: A random double-blind control clinical study. Setting: A university hospital. Patients: Forty patients with colorectal cancer, ASA I-П. Interventions: All patients were randomly divided into a DEX group (n = 20) and a control group (n = 20). Before induction of anesthesia, epidural catheters were placed in the L1-L2 or T12-L1 intervertebral spaces. The DEX group received 1 μg/kg of DEX (200 μg/50 ml) intravenously for 15 min prior to the surgery, which was then infused at a rate of 0.5 μg/kg/h until 30 min before the end of the surgery. The control group received an intravenous infusion of saline (50 ml) instead of DEX during the same periods as the DEX group. All patients received routine anesthesia and postoperative analgesia. Measurements: Blood samples from all patients were collected at the following time points: before anesthesia (T0), 24 h after surgery (T1), 48 h after surgery (T2) and 72 h after surgery (T3). Changes in T-lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+) and NK cells were determined by flow cytometry. Main Results: Compared with the control group, the percentages of CD3+ and CD4+ cells and the CD4+/CD8+ ratio in the DEX group increased significantly from T1 to T3

Effect on T cell subsets and function of isletβ cells of levemir combined with acarbose in elder patients with early-onset type 2 Diabetes Mellitus

Objective:To discuss the effect of the combined therapy of levemir and acarbose on T cell subsets and function of isletβ cells in elder patients with early-onset type 2 Diabetes Mellitus. Methods:According to the number parity of entry sequence, 100 cases of elder patients with early-onset type 2 Diabetes Mellitus are divided into the control group and the observation group of 50 cases. The control group was treated with novolin and acarbose, the observation group was given subcutaneous injection of levemir and acarbose treatment. Compare the T cell subsets and function of isletβ cells in two group of patients before the treatment (T0), treatment for 4 weeks (T1) ,treatment for 8 weeks (T2).Results:(1) The levels of T0, T1, T2CD3+, CD4+, CD4+/CD8+ were increased in both groups, and CD8+ decreased. Among them, the levels of T1, T2CD3+, CD4+, CD4+/CD8+ of the observation group were obviously higher than the control group, the level of CD8+ was lowly than the control group, the difference was statistically significant;(2) In the stage of T0, T1, T2, the levels of FPG, HbA1c, HOMA-IR were showed a downward trend, the levels of FIns, HOMA-B were increased. In these two groups, the levels of T1, T2FPG, HbA1c, HOMA-IR of the observation group were lower than the control group, and the levels of FIns, HOMA-B were higher than the control group, the difference was statistically significant;(3) In the control group occurred 3 cases of hypoglycemia, and the incidence of adverse reactions was 6%. However, in the observation group no occurred adverse reactions, the difference was statistically significant.Conclusions:The combined therapy of levemir and acarbose in elder patients with early-onset type 2 Diabetes Mellitus, It helps to improve immune function, protect the isletβ-cell function.

Study on T lymphocyte subsets and NK cells in patients with Graves' disease combined with type 2 diabetes

Objective To investigate changes in T lymphocyte subsets and NK cells in patients with simple Graves’ disease(GD)and Graves’ disease combined with type 2 diabetes mellitus(GD/T2DM).Methods Fifteen cases of GD/T2DM were selected from our hospital from November 2001 to November 2004.Before and after therapy thyroid function,thyroglobulin antibody(TGA),thyroid microsomal antibody(TMA)and blood glucose level were measured,and T lymphocyte subsets(CD3,CD4,CD8,CD4/CD8)and NK cells(CD56)were measured by immunofluorescence double labeling monoclonal antibody and flow cytometry,respectively.At the same time,comparison was made with simple GD(15 cases),T2DM(15 cases)and healthy control(20 cases).Results Before therapy,CD4/CD8,CD4 and NK cells in GD/T2DM were less than normal,and there was no significant difference in comparison with simple GD(P<0.05).In T2DM group,only CD4/CD8 and CD4 were less than those of healthy controls(P<0.05).When thyroid function recovered after 1 to 3 months of methimazole treatment in both GD/T2DM and simple GD groups,various indexes recovered,which were more obvious in simple GD.Conclusion Immune hypofunction of GD may be the key to the immune abnormality of GD/T2DM,which is more significant than that of simple GD or T2DM.The recovery of thyroid function and immune abnormality is not consistent,and the recovery of GD is more significant than that of GD/T2DM.

Relationship of serum neurotransmitters with anxiety depression, T lymphocyte subsets and NK cells in patients with lung cancer chemotherapy

Objective:To study the relationship of serum neurotransmitters with anxiety depression, T lymphocyte subsets and NK cells in patients with lung cancer chemotherapy.Methods: 56 cases of patients with advanced lung cancer who received chemotherapy in the First Affiliated Hospital of Chengdu Medical College between July 2013 and August 2016 were collected as observation group, and 50 healthy subjects who received physical examination in our hospital during the same period were selected as normal control group. Serum neurotransmitter, negative emotions and immune index levels were compared between the two groups of subjects. Pearson test was used to evaluate the relationship of serum neurotransmitter contents with negative emotions and immune index levels in patients with lung cancer chemotherapy. Results: Serum neurotransmitters DA, 5-HT and NE contents in observation group were lower than those in normal control group;SAS and SDS scores were higher than those of normal control group;peripheral blood CD4+T lymphocyte level, CD4+/CD8+ ratio and NK cell level were lower than those in normal control group while CD8+ T lymphocyte level was higher than that in normal control group. Pearson test showed that serum neurotransmitters DA, 5-HT and NE contents in patients with lung cancer chemotherapy were directly correlated with anxiety depression, T lymphocyte subset and NK cell levels.Conclusion: Serum neurotransmitter expression decrease in patients with lung cancer chemotherapy, and this is one of the important causes of anxiety depression and immune dysfunction in patients.

Detection of lymphocyte subsets and regulatory T cells in peripheral blood of patients with acute leukemia and its clinical significance

Objective To study the changes of lymphocyte subsets and regulatory T cells in peripheral blood of patients with acute leukemia(AL) and its clinical significance.Methods The different levels of peripheral blood lymphocyte subsets and regulatory T cells of 60 AL patients and 40 normal controls were detected with flow cytometry.Results Compared with the normal controls,the percentages of CD3+ T cells,CD4+ T cells,CD16+CD56+ NK cells and the ratio of CD4+ /CD8+ obviously decreased in newly diagnosed AL group(P <0.05),while their percentages of CD8+ T cells and CD19+ B cells significantly increased(P <0.01).The percentage of CD4+ T cells and the ratio of CD4+ /CD8+ in acute lymphoblastic leukemia(ALL) group were much lower than those in acute myelogenous leukemia(AML) group(P <0.01).Compared with these in control group,the proportions of CD4+ CD25high Treg cells and CD4+ CD25+ T cells in newly diagnosed AL group were significantly increased(P <0.01).Conclusion Cellular immune function is significantly abnormal in patients with AL.Compared with AML patients,ALL patients had poorer cellular immune function.The increased CD4 + CD25high Treg cells might be one of the important reasons of immunosuppression in AL.Detection of lymphocyte subsets and regulatory T cells is of clinical value on the evaluation of therapeutic effect and prognosis in AL patients.

The Effect of BCG-PSN on T-cell Subsets and Cytokines in Vernal Conjunctivitis

The effects of BCG PSN on T cell subsets and cytokines in vernal conjunctivitis were observed. The level of total IgE was quantitatively determined before and after treatment with BCG PSN by allergen diagnostic instrument in vitro . The content of T cell subsets of peripheral blood and cytokine were determined by using indirect immune fluorescence method, and IL 4 and INF γ were quantified by ELISA. The results showed that the level of total IgE was substantially reduced ( P <0.01) after treatment in the BCG PSN group. Meanwhile, CD + 8 was decreased, CD + 4 and CD + 4/CD + 8 ratio elevated with significant differences ( P <0.05) as compared with pre treatment results. The changes in total IgE, CD + 8 ,CD + 4 and CD + 4/CD + 8 ratio after treatment also presented significant differences ( P <0.05) between BCG PSN group and routine treatment group. The level of IL 4 in serum declined ( P <0.05) after treatment in the BCG PSN group, and INF γ went up ( P <0 05). IL 4 and INF γ in serum showed significant differences ( P <0.05) between two groups after treatment. It is concluded that BCG PSN has a bi directional immunoregulating effect. It can bring CD + 4 and CD + 8 into homeostasis, thereby preventing the occurrence of anaphylaxis. At the same time, BCG PSN can restrain Th 2, decrease the synthesis of IL 4, switch the balance of Th l/Th 2 to Th 1 side, boost up the predominance of Th 1 relatively, which is propitious to perennial stabilization and recovery of vernal conjunctivitis.

CD34^(+)CD38^(-)subpopulation without CD123 and CD44 is responsible for LSC and correlated with imbalance of immune cell subsets in AML

Acute myeloid leukemia(AML)is regarded as a stem cell disease.However,no one unique marker is expressed on leukemia stem cells(LSC)but not on leukemic blasts nor normal hematopoietic stem cells(HSC).CD34^(+)CD38^(-)with or without CD123 or CD44 subpopulations are immunophenotypically defined as putative LSC fractions in AML.Nevertheless,markers that can be effectively and simply held responsible for the intrinsical heterogeneity of LSC is still unclear.In the present study,we examined the frequency of three different LSC subtypes(CD34^(+)CD38^(-),CD34^(+)CD38^(-)CD123^(+),CD34^(+)CD38^(-)CD44^(+))in AML at diagnosis.We then validated their prognostic significance on the relevance of spectral features for diagnostic stratification,immune status,induction therapy response,treatment effect maintenance,and long^(-)term survival.In our findings,high proportions of the above three different LSC subtypes were all significantly characterized with low complete remission(CR)rate,high relapse/refractory rate,poor overall survival(OS),frequent FLT3^(-)ITD mutation,the high level of regulatory T cells(Treg)and monocytic myeloid^(-)derived suppressor cells(M^(-)MDSC).However,there was no significant statistical difference in all kinds of other clinical performance among the three different LSC groups.It was demonstrated that CD34^(+)CD38^(-)subpopulation without CD123 and CD44 might be held responsible for LSC and correlated with an imbalance of immune cell subsets in AML.

The influence of immunomodulator on the immunoglobulin and T cell subsets in children with intractable epilepsy

Objective:To observe the influence of immunomodulator on the immunoglobulin and T cell subsets in children with intractable epilepsy.Method:A total of 82 children with intractable epilepsy in our hospital were selected and randomly divided into 2 groups: the control group (41 cases) and the observation group (41 cases). Routine antiepileptic drugs were given to the control group. Medication regimen was that more than 2 kinds of anti epileptic drugs were be combined used. But the immunomodulator wasn't used. Treatment of immune globulin was given to the control group on the basis of observation group. By taking the 400 mg/kg/d as the standard of dosage, for 5 d every course of treatment. One course of treatment was carried out every month, 3 months in total. The changes of IgA, IgG, IgM and CD3+, CD4+, CD8+, CD4+/CD8+ were compared in 2 groups before and after treatment.Result: The comparison of IgA, IgG, IgM in the two groups before treatment was not statistically significant. After treatment, IgA, IgG in observation group were significantly higher than that before treatment and the difference was statistically significant. However, there was no significant difference on the IgM. There was no significant difference on the IgA, IgG, IgM in the control group compared with that before treatment. IgA, IgG in observation group was significant higher than that in the control group. The comparison of IgM between 2 groups was not statistically significant. The comparison of CD3+, CD4+, CD8+, CD4+/CD8+ in the two groups before treatment was not statistically significant. After treatment, CD3+, CD4+, CD4+/CD8+ in observation group were significantly higher than that before treatment;CD8+ in observation group was significantly lower than that before treatment. The difference was statistically significant. There was no significant difference on the CD3+, CD4+, CD8+, CD4+/CD8+ in the control group compared with that before treatment. CD3+, CD4+, CD4+/CD8+ in observation group was significant higher than that in the control group. CD8+ in observation group was significant lower than that in the control group. The difference was statistically significant.Conclusion:Compared with using routine antiepileptic drugs , application of immune globulin as an immunomodulator combined with conventional antiepileptic drug in children with refractory epilepsy can effectively improve the expression of IgA, IgG, IgM and T cell subsets, which has a positive effect on the immune function of the children.

Does East Meet West?—The Association between Oriental Tongue Inspection and Western Clinical Assays of White Blood Cell Subsets

The autonomic nervous system (ANS) controls white blood cell (WBC) subsets;therefore, the status of ANS can be assessed by assaying WBCs. However, this requires invasive blood sampling, time, cost, and training. Therefore, this study focused on a traditional technique, tongue inspection, which is a simpler method. The purpose of this study was to investigate whether there is an association between the traditional method of tongue inspection and clinical assay of WBC subsets. Twenty-one female alopecia areata patients were divided into two age-matched groups: 1) alopecia areata totalis (AT);and 2) alopecia areata multiplex (AM). Images of patient tongues were captured by a digital camera and categorized before blood sampling. Finally, patients were divided into five groups (normal, Yin+, Yang–, Yin– and Yang+) based on the Eight Principles of traditional Chinese medicine (TCM). Concurrently, venous blood was obtained for WBC subsets. The absolute numbers of WBCs and granulocytes of the AT group were higher than those of the AM group. The AT group was Yin+ but not Yang+, whereas the AM group was Yang+ but not Yin+. Thus, the AT group showed more elements of “cold” (Yin > Yang) compared with the AM group with elements of “hot” (Yin < Yang). Tongue inspection suggested a possibility of consistence with those of WBCs although statistical significance was not obtained. Moreover, some Yin+ and Yang+ subjects showed some trend in similarities between tongue inspection and WBC subsets although this was not statistically significant. Therefore, traditional techniques (such as tongue inspection) acupuncture must be studied further to detect whether subtle effects are induced by acupuncture treatment. As this study is underpowered, a larger scale study including males is required in the future.

In vitro incubation of cytokine-induced killer cells from patients with and without hepatitis B virus and a cell subset analysis

Objective The aim of the study was to explore the difference between immune cell subsets during the incubation of cytokine-induced kill cells （CIKs） from patients with and without hepatitis B virus （HBV）. Methods Peripheral blood samples were extracted from 50 tumor patients, and were divided into two groups according to the presence or absence of HBV. The proliferation rate and activity of CIK cells were examined based on counts on days 1, 5, 7, 9, 11, 13, and 15 of culture. Additionally, the CD3＋, CD4＋, CD8＋, CD3＋CD8＋, C＋）3＋CD4＋, and CD3＋CD56＋ T cell populations were analyzed by flow cytometry on days 5, 7, 10, 13, and 15 of culture. Results Proliferation over a 15-day period was higher in the HBV-positive group than in the negative group （280-fold vs. 180-fold increase, respectively）, but there was no significant difference between the two groups at each time point. The frequencies of CD3＋, CD8＋ T, CD3＋CD8＋, and CD3＋CD56＋T cells increased over time, while those of CD4＋ and CD3＋CD4＋ T cells decreased over time, and these changes were greater in the positive group than in the negative group. The differences in CD8＋ T cells and CD3＋CD4＋ T cells between the two groups were significant （P 〈 0.05）. Conclusion The proliferative capacity of CIK cells was higher for patients in the HBV-positive group than those in the HBV-negative group, and immune cell subsets were more favorable in the HBV-positive group than the neaative arouD.

Relationship between serum levels of IL-4,IL-8,TNF-alpha,T cell subsets and prognosis in patients with psoriasis vulgaris

Objective:To study the relationship between serum levels of IL-4,IL-8,TNF-alpha,T cell subsets and prognosis in patients with psoriasis vulgaris.Methods:A total of 120 patients with psoriasis vulgaris who were treated in our hospital from January 2018 to January 2019 were selected as the study group,and 50 normal subjects who underwent health examination in our hospital during the same period were selected as the control group.The levels of serum IL-4,IL-8,TNF-alpha and T cell subsets in the observation group and the control group were detected and compared.The levels of IL-4,IL-8,TNF-alpha and T cell subsets in the observation group at different time after treatment were compared after standardized western medicine treatment.Pearson test was used to analyze the correlation between IL-4,IL-8,TNF-alpha and T cell subsets.Results:The levels of IL-4,IL-8 and TNF-a in the observation group were higher than those in the control group,while the levels of CD3+,CD4+,CD4+/CD8+in the observation group were lower than those in the control group,and the levels of CD8+in the observation group were higher than those in the control group.There was a significant difference between the two groups.In the observation group,after 8 weeks of treatment,the levels of IL-4,IL-8 and TNF-alpha continued to decrease,CD3+,CD4+,CD4+/CD8+increased and CD8+decreased with the prolongation of treatment time.There was significant difference among the groups.Pearson correlation test was used.IL-4,IL-8,TNF-a had negative correlation with CD3+,CD4+,CD4+/CD8+,and positive correlation with CD8+.Conclusion:The incidence of psoriasis vulgaris is related to the elevation of IL-4,IL-8,TNF-alpha levels and immunodeficiency.The prognosis of psoriasis vulgaris can be judged by monitoring the levels of IL-4,IL-8,TNF-alpha and T cell subsets.

Effects of combined treatment of bronchial arterial chemoembolization and radioactive particle implantation on tumor markers and T lymphocyte subsets in locally advanced non-small cell lung cancer

Objective: To investigate the effects of bronchial arterial chemoembolization combined with radioactive particle implantation on the level of serum tumor markers and T lymphocyte subsets in patients with locally advanced non-small cell lung cancer. Methods: A total of 91 cases of locally advanced non-small cell lung cancer patients according to the random data table were divided into the control group (n=45) and observation group (n=46) according to the random data table. Patients in the control group was treated with bronchial arterial chemoembolization, on the basis of the control group, patients in the observation group were treated with radioactive particle implantation, the serum tumor markers and T lymphocyte subsets of the two groups were compared before and after treatment. Results: The levels of CEA, NSE, CA125, CD4+, CD8+, CD4+/CD8+ and NK in the two groups before the treatment were not statistically significant. Compared with the group before treatment, levels of CEA, NSE, CA125and CD8+ of the two groups after treatment were significantly decreased, and after treatment the level of CEA, NSE, CA125and CD8+ in the observation group was significantly lower than those of the control group;The levels of CD4+, CD4+/CD8+ and NK in the two groups after treatment were significantly higher than those in the group before treatment, and the observation group levels were significantly higher than those of the control group. Conclusion: Bronchial artery embolization combined with radioactive particle implantation for locally advanced non-small cell lung cancer, can effectively reduce the serum tumor markers level, improve the level of T cell subsets of patients, has important clinical value.

Increased CD4^+CD45RA^-FoxP3^(low) cells alter the balance between Treg and Th17 cells in colitis mice

AIM To investigate the role of regulatory T cell(Treg) subsets in the balance between Treg and T helper 17(Th17) cells in various tissues from mice with dextran sulfate sodium-induced colitis.METHODS T r e g c e l l s, T r e g c e l l s u b s e t s, T h 1 7 c e l l s, a n d CD4+CD25+FoxP 3+IL-17+ cells from the lamina propria of colon(LPC) and other ulcerative colitis(UC) mouse tissues were evaluated by flow cytometry. Forkhead box protein 3(FoxP 3), interleukin 17A(IL-17A), and RORC m RNA levels were assessed by real-time PCR, while interleukin-10(IL-10) and IL-17 A levels were detected with a Cytometric Beads Array.RESULTS In peripheral blood monocytes(PBMC), mesenteric lymphnode(MLN), lamina propria of jejunum(LPJ) and LPC from UC mice, Treg cell numbers were increased(P < 0.05), and FoxP 3 and IL-10 mR NA levels were decreased. Th17 cell numbers were also increased in PBMC and LPC, as were IL-17 A levels in PBMC, LPJ, and serum. The number of FrI subset cells(CD4+CD45RA+FoxP 3low) was increased in the spleen, MLN, LPJ, and LPC. FrI I subset cells(CD4+CD45RA-Fox P3high) were decreased among PBMC, MLN, LPJ, and LPC, but the number of Fr III cells(CD4+CD45RA-FoxP 3low) and CD4+CD25+FoxP 3+IL-17A+ cells was increased. Fox P3 m RNA levels in CD4+CD45RA-Fox P3 low cells decreased in PBMC, MLN, LPJ, and LPC in UC mice, while IL-17 A and RORC mR NA increased. In UC mice the distribution of Treg, Th17 cells, CD4+CD45RA-FoxP 3high, and CD4+CD45RA-FoxP 3low cells was higher in LPC relative to other tissues.CONCLUSION Increased numbers of CD4+CD45RA-FoxP 3low cells may cause an imbalance between Treg and Th17 cells that is mainly localized to the LPC rather than secondary lymphoid tissues.

Advanced Glycation End Products Promote Differentiation of CD4^+ T Helper Cells toward Pro-inflammatory Response

This study investigated the effect of advanced glycation end products（AGEs） on differentiation of na ve CD4＋T cells and the role of the receptor of AGEs（RAGE） and peroxisome proliferator-activated receptors（PPARs） activity in the process in order to gain insight into the mechanism of immunological disorders in diabetes. AGEs were prepared by the reaction of bovine serum albumin（BSA） with glucose. Human na ve CD4＋T cells, enriched from blood of healthy adult volunteers with negative selection assay, were cultured in vitro and treated with various agents including AGEs, BSA, high glucose, PGJ2 and PD68235 for indicated time. In short hairpin（sh） RNA knock-down experiment, na ve CD4＋T cells were transduced with media containing shRNA-lentivirus generated from lentiviral packaging cell line, Lent-XTM293 T cells. Surface and intracellular cytokine stainings were used for examination of CD4＋T cell phenotypes, and real-time PCR and Western blotting for detection of transcription factor mRNA and protein expression, respectively. The suppressive function of regulatory T（Treg） cells was determined by a [3H]-thymidine incorporation assay. The results showed that AGEs induced higher pro-inflammatory Th1/Th17 cells differentiated from na ve CD4＋T cells than the controls, whereas did not affect anti-inflammatory Treg cells. However, AGEs eliminated suppressive function of Treg cells. In addition, AGEs increased RAGE mRNA expression in na ve CD4＋T cells, and RAGE knock-down by shRNA eliminated the effect of AGEs on the differentiation of CD4＋T cells and the reduction of suppressive function of Treg cells. Furthermore, AGEs inhibited the mRNA expression of PPARγ, not PPARα; PPARγ agonist, PGJ2, inhibited the effect of AGEs on na ve CD4＋T cell differentiation and reversed the AGE-reduced suppressive function of Treg cells; on the other hand, PPARγ antagonist, PD68235, attenuated the blocking effect of RAGE shRNA on the role of AGEs. It was concluded that AGEs may promote CD4＋T cells development toward pro-inflammatory state, which is associated with increased RAGE mRNA expression and reduced PPARγ activity. ＋

Recovery of natural killer cells is mainly in post-treatment period in chronic hepatitis C patients treated with sofosbuvir plus ledipasvir

AIM To investigate how natural killer(NK) cells are affected in the elimination of hepatitis C virus(HCV) by sofosbuvir/ledipasvir, two highly effective direct-acting antivirals(DAAs). METHODS Thirteen treatment-na?ve and treatment-experienced chronic hepatitis C(CHC) patients were treated with sofosbuvir/ledipasvir, and NK cells were detected at baseline, weeks 2, 4, 8 and 12 during therapy, and week post of treatment(Pt)-12 and 24 after the end of therapy by multicolor flow cytometry and compared with those from 13 healthy controls. RESULTS All patients achieved sustained virological response. There was a significant decline in CD56^(bright) NK cell frequencies at week 8(P = 0.002) and week 12(P = 0.003), which were altered to the level comparable to healthy controls at week Pt-12, but no difference was observed in the frequency of CD56^(dim) NK cells. Compared with healthy controls, the expression levels of NKG2A, NKp30 and CD94 on NK cells from CHC patients at baseline were higher. NKG2A, NKp30 and CD94 started to recover at week 12 and reached the levels similar to those of healthy controls at week Pt-12 or Pt-24. Before treatment, patients have higher interferon(IFN)-γ and perforin levels than healthy controls, and IFN-γ started to recover at week 8 and reached the normalized level at week Pt-12. CONCLUSION NK cells of CHC patients can be affected by DAAs, and phenotypes and function of NK cells recover not at early stage but mainly after the end of sofosbuvir/ledipasvir treatment.

Cancer and Germ Cells

Background: Era of contraception, abortions, [20th, 21st centuries] implemented as family welfare schemes witnessed, increased global incidence of cancer, tumors, neoplasm and mortality. Objectives: Altruistic association of contraception [if any], with increasing cancer, tumor was sought after. Methods: In 2012, retrospective analysis of, prevalence of cancer, tumor in 350 patients of 20 - 35 years, 35 - 50 years, >50 years age groups, from data collected by convenient, stratified random sampling, from different geographical locations, between 2002-2012 and its association with presence, absence of contraception, abortion was undertaken;simultaneously, serum estrogen levels obtained from 105 patients, were also analyzed. From 1983-2012 clinical practice, 212 patients treated for different types of neoplasm namely breast cancer, prostate cancer, cancer cervix and benign prostatic hyperplasia were randomly allotted to the above 3 age groups and the data were analyzed for association with contraception status and possible significance. Results: 6 fold increase in cancer incidence was seen in contraceptive users among >50 years with a p value of <0.0005. Contraception was associated with 4 - 7 fold increase in tumor prevalence among >35 - >50 years with a p value of <0.0005. Endogenous estrogen had decreased to ~5-8 pg in 61% of contraceptive users with a p value of <0.0005;after hysterectomy endogenous estrogen values up to ~0.4 pg were seen. Cholesterol deprived diet, due to decreased synthesis of endogenous estrogen:androgen also was associated with 50% increase in tumor, cancer in youth without contraception. Estrogen receptors were positive in well differentiated cancers of breast, associated with reduced levels of endogenous estrogen among contraceptive users, suggesting estrogen receptor positivity could be a compensatory phenomenon;anaplastic tumors did not exhibit estrogen receptor positivity. 10 - 20 fold increase in breast cancer was seen among 20 - >50 years, in contraceptive users with a p value of <0.0005;20 - 30 fold increase in prostate cancer was seen among 35 - >50 years, in contraceptive users with a p value of <0.0005;cancer of the cervix had increased 20 - 40 fold, among 20 - 70 years, in contraceptive users with a p value of <0.0005. Conclusion: Concept is acquired contraception preventing traversal of normal path by germ cells with resultant smashed destruction of germ cells, consequent reduced endogenous estrogen:androgen surveillance, leading to agonizing faults of cellular genomic repertoire, uncontrolled multiplication preceded by no differentiation of cell cycle, metabolism, resulting in soaringly high incidence of cancers including breast, prostate and uterine cervix in both life partners. Increased estrogen receptors:androgen receptors in breast, prostate well differentiated cancers respectively are, probably a compensatory rise, secondary to the sudden artificially acquired contraception resulting in significant reduction of endogenous germ cell hormones in contraceptive users. Contraception reversal with chemotherapy, radiation therapy, surgery achieves arrest of progression of cancer, reduces incidence, prevalence of neoplasm, as a cause-effect phenomenon and not castration or anti estrogen:antiandrogens which will perpetuate, promote neoplastic diseases by decreasing endogenous estrogen:androgen.

CD4^+CD25^(high) Regulatory Cells in Peripheral Blood of NSCLC Patients

The proportion and changes of CD4^＋CD25^high regulatory T cells （Trs） in peripheral blood of non-small cell lung cancer （NSCLC） patients were analyzed and their clinical significance explored. The peripheral blood was collected from 61 patients with NSCLC and 15 healthy controls. By using monoclonal antibodies, the blood samples were evaluated with the flow cytometry for lymphocyte subsets （CD3^＋, CD4^＋ and CD8^＋） and CD4^＋CD25^high Tr cells. The results showed that the proportion of CD4^＋CD25^high Tr cells in NSCLC group was significantly higher than in control group [（4.36 ±2.07） % vs （2.04±1.03） %, P〈0.01]. The proportion of CD4^＋CD25^ high Tr cells in late stage was higher than that in early stage [stages Ⅰ ＋Ⅱ （2.264±0.6） %; stage Ⅲ（3.284± 1.38） %; stage IV （6.06 4±4.08） %] （P〈0.05）. Kaplan-Meier survival analysis revealed that the prognosis of the patients who had higher proportion of CD4^＋CD25^high Tr cells in peripheral blood was worse （P=0.0026）. In conclusion, the relative increase in CD4^＋CD25^high Tr cells in peripheral blood may be related to im- munosuppression and tumor progression in patients with NSCLC. This finding suggests that CD4^＋CD25^high Tr cells in peripheral blood of NSCLC may be positive for prognosis analysis. The use of depletion of the CD4^＋CD25^high Tr cell therapy to treat NSCLC patients may be an effective strategy.

Immunohistochem ical Study of T Lym phocyte Subsets onFrozen Substituted Plastic Em bedding Section of the BoneMarrow from Patients with Myelodysplastic Syndrom e andIts ClinicalIm plication

An immunohistochemical study of T lymphocyte subsets on frozen substituted plastic embedding bone marrow sections obtained from 10 patients with myelodysplastic syndrome (MDS) was presented. The results of qualitative and quantitative immunohistochemical analysis are as follows: (1) Labile antigens of T lymphocytes were well preserved, thus allowing analysis of distribution of T lymphocyte subsets in situ ; (2) the average number of T 3, T 4 and T 8 lymphocyte of the diffuse infiltrate was about 2 %, 0.4 %, 0.5 %, respectively, of all nucleated cells in bone marrow, and T 4/T 8 of T cells were below 1.0 in patients with MDS; (3) there were cases of RAS showing T lymphocyte aggregation in bone marrow, but no patient exhibited progressive refractory anemia with excess of blasts(RAEB) and RAEB in transformation (RAEBT). These findings indicated that the immunological abnormalities are of importance in the evaluation of pathogenesis and prognosis of MDS.