Changes in the proportion of lymphocyte subsets and the expression of surface receptors in peripheral blood of patients with colorectal cancer / 细胞与分子免疫学杂志

Objective To identify the sets of lymphocytes that could systematically evaluate immune function of colorectal cancer patients, based on the expression of colorectal cancer T cells, natural killer (NK) cells, and NKT cell surface protein receptors. Methods Peripheral blood samples from 144 patients with colorectal cancer and 87 healthy controls were collected, and the differences in surface receptors of lymphocyte subsets in peripheral blood of patients and healthy controls were analyzed by means of flow cytometry and cell culture. Results Compared with healthy control group, the percentage of peripheral blood total lymphocytes, CD16brightCD56dimNK cells and NKT cells decreased in patients with colorectal cancer. The percentage of T cells, CD16brightCD56dimNK cells and NKT cell surface inhibitory receptors T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitor motif domains (TIGIT) increased; T cells, NK cells, NKT cell surface chemokine receptor C-C motif chemokine receptor 7 (CCR7) slightly decreased. Conclusion There are differences in the proportion of NK cell subsets and the expression profile of surface receptors in peripheral blood of patients with colorectal cancer.

Screening of a novel CRM1 targeting inhibitor and its effects on the proliferation and growth of extranodal NK/T cell lymphoma / 中国药科大学学报

@#The purpose of this study was to screen out the novel chromosome maintenance protein 1(CRM1)covalent targeting inhibitors by computer-assisted drug design(CADD), and to study their effects on the proliferation of extranodal nature killer/T cell lymphoma(ENKTL). A novel CRM1 inhibitor LFS-829 was designed based on the molecular structure of LFS-01 by means of ADME/T and covalent docking. The target binding of LFS-829 with CRM1 was analyzed by MALDI-TOF mass spectrometry. The effects of LFS-829 on the proliferation of extranodal NK/T cell lymphoma SNK6 and HANK-1 cells were detected by CCK-8. The cell morphology was observed by live cell workstation. Immunofluorescence experiments were used to analyze the effect of LFS-829 on nuclear export function of CRM1. The changes of NF-κB signaling pathway under different concentrations of LFS-829 were analyzed by Western blot, dual luciferase reporter gene assay and enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry, and the expression of proteins related to apoptosis pathway was detected by Western blot. Tests of peripheral blood mononuclear lymphocyte(PBMC)toxicity, platelet toxicity and mouse acute toxicity were done to make sure that it is not harmful to human. LFS-829 could bind covalently to the cysteine residue of the hydrophobic active pocket of CRM1. LFS-829 could selectively kill SNK6 and HANK-1 cells, with IC50 of 366 nmol/L and 158 nmol/L in 72 h, respectively, and cell morphology was significantly changed. LFS-829 at 800 nmol/L significantly inhibited the nuclear export function of CRM1, promoted nuclear assembly of IκB-α, down-regulated the transcriptional activity of NF-κB signaling pathway, significantly up-regulated the expression of apoptotic pathway protein p53, cleaved Caspase 3 and cleaved Caspase 9 and induced apoptosis, with no obvious killing effect on PBMC or platelets. It did not cause substantial tissue damage to mice at the high dose of 300 mg/kg, which shows its great prospect of future application.

Expression of death receptor 5 upreguleted by chloroquine enhances the sensitivity of Huh7 cells to tumor necrosis factor related apoptosis-inducing ligand / 肿瘤研究与临床

Objective To explore the effect of chloroquine on death receptor 5 (DR5) expression of hepatocellular carcinoma Huh7 cells and cell proliferation and apoptosis induced by tumor necrosis factor related apoptosis-inducing ligand (TRAIL).Methods Huh7 cells were divided into four groups:the control group (1∶1 000 dimethyl sulfoxide),TRAIL group (50 μg/L),chloroquine group (10 μmol/L) and TRAIL +chloroquine group (TRAIL 50 μg/L + chloroquine 10 μmol/L).Thiazolyl blue tetrazolium bromide (MTT) assay was used to determine the proliferation activity of cells,immunofluorescence was used to detect the expression of DR5,4',6-diamidino-2-phenylindole (DAPI) staining was used to observe cell apoptosis and Western blot was used to detect the expression of cleaved poly ADP-ribose polymerase (PARP).Results TRAIL treatment could decrease Huh7 cells proliferation activity;when compared with the cell viability in the control group,the cell proliferation inhibition rate of chloroquine group,TRAIL group and TRAIL+ chloroquine group was (89±8) ％,(53±10) ％ and (27±7) ％,respectively;compared with TRAIL group alone,cell proliferation activity was decreased in TRAIL+ chloroquine group (t =3.922,P =0.017).The expression of DR5 was upregulated in chloroquine group,and the cell apoptosis signaling was activated in TRAIL + chloroquine group.The cell apoptosis rate of TRAIL group and TRAIL + chloroquine group was (10.0±2.3) ％ and (20.4±4.0) ％,respectively,and there was a statistical difference (t =3.894,P =0.018).Conclusion Chloroquine can enhance the cell chemosensitivity to TRAIL treatment by upregulating the expression of DR5 in Huh7 cells.

Naturally derived sulforaphene inhibits proliferation of mantle cell lymphoma by down-regulating CRM1 expression / 肿瘤

Objective: To investigate the expression level of chromosomal region maintenance 1 (CRM1) in mantle cell lymphoma, and to explore the possible mechanism of naturally derived sulforaphene LFS-01 inhibiting the proliferation of mantle cell lymphoma cells. Methods: The expression of CRM1 in mantle cell lymphoma was analyzed by Oncomine data mining. After treatment with LFS-01, the viability of mantle cell lymphoma JeKo-1 cells was detected by CCK-8 assay, the nuclear transport function of CRM1 protein was measured by laser confocal microscopy, the expression of CRM1 protein was detected by Western blotting, the cell cycle arrest and apoptosis were analyzed by FCM and transmission electron microscopy, the expression change of apoptosis pathway-related proteins was detected by Western blotting, and the impact of caspase inhibitor z-VAD-FAM on the effects of LFS-01 was detected by CCK-8 assay and FCM. Lentiviral infection was used to establish a stable JeKo-1 cells expressing mutant CRM1 (C528S), then the effects of LFS-01 on the nuclear transport function of CRM1 and the proliferation of JeKo-1 cells were detected by laser confocal microscopy and CCK-8, respectively. The transcriptional level in JeKo-1 cells after LFS-01 treatment was detected by RNAseq, and the viability of JeKo-1 cells treated with Toll-like receptor (TLR) inhibitor TAK-242 and LFS-01 was measured by CCK-8 assay. Results: CRM1 was overexpressed in mantle cell lymphoma (P < 0.05). LFS-01 inhibited the proliferation of JeKo-1 cells, and the 50% inhibitory concentration (IC50) at 24 h and 48 h were 5.81 μmol/L and 9.09 μmol/L, respectively. 20.0 μmol/L LFS-01 inhibited the nuclear transport function of CRM1 (P < 0.05), 6.0 μmol/L LFS-01 down-regulated the expression level of CRM1 (P < 0.05), 10.0 μmol/L LFS-01 induced cell cycle arrest at G2/M phase and induced apoptosis (both P < 0.05), and 4 μmol/L LFS-01 up-regulated the expression levels of poly ADP-ribose polymerase (PARP), cleaved caspase-3 and cleaved caspase-9 (all P < 0.01). The caspase inhibitor z-VAD-FAM reversed the apoptosis-induction effect of LFS-01 (P < 0.01). CRM 1 mutation eliminated the effects of LFS-01 on CRM1 function and cell proliferation (both P < 0.05). LFS-01 inhibited the cell proliferation-related pathways (P < 0.01), and TLR inhibitor TAK-242 combined with LFS-01 had synergetic inhibitory effect on JeKo-1 cells (P < 0.01). Conclusion: LFS-01 can inhibit the growth of mantle cell lymphoma cells through inducing cell cycle arrest and apoptosis, and CRM1 is essential in this process. In addition, LFS-01 and TLR inhibitor TAK-242 have synergetic inhibitory effect on mantle cell lymphoma cells.

Effect of Chinese herb acupoint application on constipation in advanced cancer patients / 现代临床护理

Objective To explore the effect of acupoint application with Chinese herbs on constipation in advanced cancer patients. Methods Sixty-one advanced cancer patients with constipation were randomly divided into the experiment and the control groups with 31 in the experiment group and 30 in the control group. The control group was treated only by routine care , while the experiment group was cared by routine nursing points and acupoint application with Shenque and Zusanli , 6 h perday for 7 d. The two groups were compared in terms of the curative effect. Result The response rates in the experiment and control groups were 87.1%and 56.7%, respectively (P<0.05). Conclusion Acupoint application with Chinese herbs to treat constipation in patients with advanced tumors has a better therapeutic effect.

Clinical efficacy and safety of hysteroscopy surgery combined with methotrexate for treating the patients with cesarean scar pregnancy / 中国医师进修杂志

Objective To investigate the clinical efficacy and safety of hysteroscopy surgery combined with methotrexate (MTX) for treating the patients with cesarean scar pregnancy(CSP).Methods The clinical data of 64 patients with CSP were retrospectively analyzed.The patients were divided into observation group (46 cases) and control group (18 cases) according to treatment methods,the control group was given the therapy of local pregnancy intracapsular MTX combined with uterine curettage,and the observation group was given the therapy of local pregnancy intracapsular MTX combined with lesions removed surgery by hysteroscopy.The intraoperative blood Ioss,epistrophy time of β-human chorionic gonadotrophin (hCG),hospitalization time,clinical efficacy of the two groups were observed.Follow-up for 6 months the menstruation of the two groups was observed.Results The total effective rate in observation group was significantly higher than that in control group [100.0％ (46/46) vs.66.7％ (12/18),x2 =16.920,P ＜ 0.01].The intraoperative blood loss,epistrophy time of β-hCG and hospitalization time in observation group were significantly lower than those in control group [(35.46 ± 11.63) ml vs.(176.78 ± 57.96) ml,(21.04 ± 6.79)d vs.(39.65 ± 12.79) d,(13.89 ±4.63) d vs.(26.82 ±8.94) d,P＜0.01].The observation group had a successful surgery and no complications.In the control group,4 cases occurred uterine curettage intraoperative bleeding,2 cases underwent open lesion resection and then were forced to line hysterectomy.The rate of menstruation in observation group after operation 30-39 d was significantly higher than that in control group [84.8％ (39/46) vs.25.0％ (4/16),P ＜ 0.01].Conclusions Hysteroscopy surgery combined with MTX for treating the patients with CSP has exact clinical effect.It is safe and reliable,and is worthy of promotion and application.

Immobilized enzyme reactors in HPLC and its application in inhibitor screening: A review / 药物分析学报

This paper sets out to summarize the literatures based on immobilized enzyme biochromatography and its application in inhibitors screening in the last decade.In order to screen enzyme inhibitors from a mass of compounds in preliminary screening,multi-pore materials with good biocompatibility are used for the supports of immobilizing enzymes,and then the immobilized enzyme reactor applied as the inmobilized enzyme stationary phase in HPLC.Therefore,a technology platform of high throughput screening is gradually established to screen the enzyme inhibitors as new anti-tumor drugs.Here,we briefly summarize the selective methods of supports,immobilization techniques,co-immobilized enzymes system and the screening model.

Clinical efficacy of hysteroscope operation treatment of uterine cavity adhesion and correlative risk factors analysis of uterine cavity adhesion / 中国医师进修杂志

Objective To study the correlative risk factors of uterine cavity adhesion (UCA),and observe the clinical efficacy of hysteroscope operation treatment of UCA.Methods According to whether the occurrence of UCA,120 patients with spontaneous abortion were divided into two groups.The correlative risk factors of UCA were analyzed by multiple factors regression analysis.The patients with UCA were treated with hysteroscope operation treatment,and the clinical efficacy were observed.Results In 120 patients with spontaneous abortion,46 patients (38.3％) had UCA.Single factor analysis result showed:UCA was correlated with pelvic inflammation,gravidity,uterine cavity suction negative pressure,uterine cavity suction time (P ＜ 0.05),but UCA was not correlated with age,years of education,occupation,gestational weeks,menarche age (P＞ 0.05).Multiple factors analysis result showed:UCA was correlated with pelvic inflammation,uterine cavity suction negative pressure,uterine cavity suction time (P ＜ 0.05).The total effective rates of hysteroscope operation treatment of mild,moderate and severe UCA were 100.0％ (18/18),93.8％ (15/16)and 83.3％ (10/12) respectively,there was no significant difference among them (P ＞ 0.05).Conclusions Along with the increasing of uterine cavity suction negative pressure and extension of uterine cavity suction time,the risk of UCA is also increasing,pelvic inflammation is also an independent risk factor of UCA.Hysteroscope operation treatment of UCA has better curative effect,especially for mild UCA,it is worth clinical application.

Clinical Value of Laparoscopic Supracervical Hysterectomy / 中国微创外科杂志

Objective To explore the clinical value of laparoscopic supracervical hysterectomy (LSH). Methods From July 2006 to September 2007,78 cases of LSH and 59 cases of abdominal supracervical hysterectomy (ASH) were performed in our hospital. The clinical data of the patients,including intraoperative blood loss,operation time,recovery time of bowel movement,postoperative time of out-of-bed activity and postoperative hospital stay,were analyzed and compared between the two groups.Results No significant difference was found on the mean intraoperative blood loss between the LSH group and ASH group [(65.1?25.5) ml vs (72.9?23.6) ml,t=-1.830,P=0.069]. Whereas,the LSH group had significantly longer operation time and earlier recovery of the gastrointestinal function [(80.3?29.6) min vs (62.4?13.1) min,t=4.332,P=0.000;and (26.5?8.5) h vs (30.9?7.0) h,t=-3.232,P=0.001]. Furthermore,the LSH patients had out-of-bed activity and were discharged from hospital significantly earlier than the ASH group [(32.8?6.7) h vs (40.4?9.7)h,t=-5.421,P=0.000;and (7.1?0.6) d vs (7.9?0.5) d,t=-8.291,P=0.000]. No major complication occurred in both the groups. Conclusions LSH shows great advantages over ASH. As long as surgeons are skilled in laparoscopic operation,LSH can be an ideal procedure for hysterectomy.