[Hemolytic uremic syndrome complicated with IgA nephropathy: a clinicopathological study].

Objective: To investigate the clinicopathologic characteristics, treatments, outcomes and mechanisms of hemolytic uremic syndrome (HUS) complicated with IgA nephropathy (IgAN). Methods: The clinical manifestations, treatments, prognosis and histopathological features of renal biopsy tissues were analyzed in two cases of HUS complicated with IgAN from Beijing Children's Hospital, Capital Medical University using light microscopy, immunofluorescence detection and electron microscopy. The related literatures were also reviewed. Results: The clinical manifestations were microvascular hemolytic anemia, thrombocytopenia, acute renal impairment with hematuria, proteinuria, and positive anti-H factor antibody. Histological findings confirmed presence of both HUS and IgAN. Histological features included glomerular mesangial and stromal hyperplasia with endothelial cell proliferation, capillary stenosis, arteriolar thickening, and glomerular ischemia and capillary dilatation. Immunofluorescence detection showed diffuse IgA deposition in the glomerular mesangial matrix. Electron microscopy showed proliferation of mesangial and endothelial cells, thickening of the inner layer of the glomerular basement membrane, deposition of massive electronic densification in the mesangial region, and shrinkage of the segmental basement membrane. The two children were very responsive to plasma exchange and steroid treatments. However, their urine protein and occult blood tests remained continuously positive during the follow-up of 5 years 7 months and 8 months respectively. Conclusions: HUS complicated with IgAN is rare. The diagnosis relies on various pathological examinations, which require the combination of light microscopy, immunofluorescence detection and electron microscopy. Plasma exchange and steroid treatments are effective. However, the long-term prognosis is concerning and may relate to pathological grade and secondary factors. The mechanism of connecting HUS and IgAN is unknown, but may be caused by prodromal or secondary factors.

KLHL22 promotes malignant melanoma growth in vitro and in vivo by activating the PI3K/Akt/mTOR signaling pathway.

The kelch like family member 22 (KLHL22) is a member of the KLHL (Kelch-like) gene family, which was involved in the progression of breast cancer. However, its role remains unclear in malignant melanoma (MM). Our study found that KLHL22 expression was upregulated in human MM tissues. Regarding the functional analysis for KLHL22 in the progression of MM cells, we demonstrated that overexpression of KLHL22 could promote MM cell growth in vitro. Vice versa, knockdown of KLHL22 could suppress the proliferation of MM cells. Furthermore, KLHL22 also promoted tumorigenesis of MM cells in vivo. In experiments investigating the underlying mechanism, expressions of p-Akt and p-mTOR were significantly increased by overexpression of KLHL22. Meanwhile, knockdown of KLHL22 could decrease the expression levels of p-Akt and p-mTOR. Our studies thus suggest that KLHL22 can promote the growth of MM cells via activating the PI3K/Akt/mTOR signaling pathway, which can serve as a potential target in the diagnosis and/or treatment of MM.

[Implementation of Beijing Tobacco Control Regulation in restaurants in Dongcheng and Chaoyang districts of Beijing, 2018].

Objective: To evaluate the implementation effect of the Beijing Tobacco Control Regulation. Methods: An observational study was conducted in a multi-stage randomly selected sample of 93 restaurants in Dongcheng and Chaoyang districts, Beijing. Undercover visits to the restaurants were paid by investigators at lunch or dinner time. The incidence of smoking behavior and the posters of no-smoking signs were observed, waiters were interviewed about awareness of the regulation, and comparisons with the baseline data of 6 months before and 1 month after regulation implementation were made. Results: The pasting rate of no-smoking signs was 76.3%. The awareness of the regulation in the waiters surveyed was high. The incidence rate of smoking in restaurants (29.0%) was lower than that before the regulation implementation (36.7%), but it was significantly higher than that one month after regulation implementation (14.8%). No active interventions from the restaurant staff were observed when smoking occurred. The incidence of smoking in restaurants within commercial buildings (3.3%) was significantly lower than that in non-commercial buildings (41.3%), the difference was significant (P<0.05). Conclusions: The effect of the regulation weakened 3 years after implementation compared with that in 1 month after the implementation. The enforcement degree of the regulation was conflicted with pasting rate of no-smoking signs and the regulation awareness level in waiters in restaurants in Dongcheng and Chaoyang districts.

[Prenatal and postnatal ultrasound assessment and clinical prognostic analysis for Ebstein anomaly].

Objective: To investigate the echocardiographic features of fetal Ebstein's anomaly (EA) and to analyze its clinical outcome and prognosis. Methods: A retrospective case-control study was conducted to analyze the echocardiographic features in fetus with EA. Thirty-five EA fetuses (EA group) and 35 normal fetuses matched for gestational age (control group) were enrolled. The main echocardiographic parameters of the two groups were collected and compared. According to the direction of blood flow in the ductus arteriosus (DA),fetuses in EA group were divided into DA reverse perfusion subgroup (n=11) and normal DA blood flow subgroup (n=24). The echocardiographic parameters and GOSE scores were compared between the two subgroups. The echocardiographic features of EA and the difference of fetal hemodynamics were summarized,and the clinical outcome of EA fetus was evaluated by GOSE score. Chi-square test Rank sum test or t test were used for comparison between groups. Results: Nineteen (54%) of the 35 patients terminated the pregnancy and 16 (46%) continued pregnancy until delivery during follow-up. Compared with the normal fetus group,the cardiothoracic ratio was significantly higher (0.47±0.11 vs. 0.34±0.01, t=6.640, P<0.01) and the transverse diameter ratio of right atrium to left atrium was significantly greater (1.42±0.38 vs. 1.08±0.11, t=5.030, P<0.01) in the EA group, and the ratio of pulmonary artery diameter to aortic diameter was significantly lower in the EA group (1.04±0.21 vs. 1.20±0.15, t=-3.770, P<0.01). Compared with the normal DA blood flow subgroup,the GOSE scores ≥ 1.5 was more frequently seen (7/11 vs. 3/24, P=0.004) and the ratio of pulmonary artery diameter to aortic diameter was significantly lower (0.91±0.18 vs. 1.24±0.20, t=-4.696, P=0.002) in the DA reverse perfusion subgroup. Six of the 16 delivery cases underwent EA corrective surgery after birth with 100% successful rate of operation. Among the 6 cases,two had preoperative GOSE score of ≥1.5 who were considered as severe cases,and four had low GOSE score of<1.5. The remaining ten cases who had not undergone the corrective surgery were followed up routinely. Conclusion: Prenatal hemodynamics of EA combined with GOSE score can more accurately evaluate the severity and prognosis of fetal EA, reduce unnecessary labor induction, and improve postpartum cure rate and clinical outcomes.

[Survey of exposure to second-hand smoke in residents aged 15 years and over one year after implementation of tobacco control regulation in public places in Beijing].

Objective: To monitor the second-hand smoke (SHS) exposure in residents aged 15 years and over in public venues, indoor workplaces, on public transportation vehicles and at home in Beijing and evaluate the effect of Beijing Tobacco Control Regulation. Methods: Data from 2014 and 2016 Beijing Adult Tobacco Survey were used. The surveys covered 16 districts in Beijing. The study subjects were selected through multi-stage cluster sampling with probability proportional to population size, and data were collected by using electronic questionnaire in face-to-face household interviews. A total of 8 484 and 9 372 valid questionnaires were collected for the surveys in 2014 and 2016, respectively. Statistical packages SPSS 20.0 and R 3.4.4 were used for data analyses. After weighting the samples using complex survey designs, the SHS exposure rates in different places in adults of Beijing were estimated. χ(2) tests were performed for the comparison. Results: The SHS exposure rates of residents aged 15 years and over in Beijing who visited health care facilities, government buildings, universities, primary and secondary schools and restaurants declined from 12.8%, 19.7%, 24.3%, 32.8% and 65.7% in 2014 to 6.2%, 10.8%, 12.5%, 19.1% and 32.5% in 2016, respectively. The SHS exposure rates in bars/nightclubs were 89.5% in 2014 and 80.3% in 2016. From 2014 to 2016, the SHS exposure rates declined from 35.7% to 20.0% in indoor workplaces and declined from 3.9% to 2.5% on public transportation vehicles. The SHS exposure rates at home were 39.8% in 2014 and 37.6% in 2016, respectively. Conclusions: The SHS exposure rates in public places declined obviously in Beijing after the one year implementation of Beijing Tobacco Control Regulation, indicating the effect of the regulation implementation.

[Clinical and genetic manifestations of immunodeficiency, centromeric instability, and facial anomalies syndrome: a case report and literature review].

Objective: To analyze the clinical and genetic features of immunodeficiency, centromeric instability, and facial anomalies (ICF) syndrome with a case report and literature review. Methods: The clinical data and genetic test of a girl diagnosed with ICF syndrome in the Department of Nephrology and Immunology in Qingdao Women and Children's Hospital in December 2016 were extracted and analyzed. "ICF syndrome" "immunodeficiency, centromeric instability and facial anomalies syndrome" "ICF syndrome and DNMT3B" were used as key words to search Chinese databases and Pubmed for literature until March 2018, and the literature was reviewed. Results: A female patient aged 22 months old with ocular hypertelorism and low-set ears was admitted due to recurrent infection over one year. Laboratory tests showed humoral immune deficiency with IgG<1.34 g/L, IgA<0.060 g/L, and IgM<0.179 g/L, but normal cellular immunity (total T lymphocyte 0.503, hepler T lymphocyte 0.328, cytotoxic T lymphocyte 0.166, natural killer cell 0.184, total B lymphocyte 0.276). Whole-exome sequencing revealed a de novo heterozygous splice site mutation c.922-2A>G in intron 8, and a de novo heterozygous missense mutation c.2477G>A in exon 23 of DNMT3B gene. Chromosome karyotype analysis showed 46, XX, with 64 out of 100 karyotypes showing centromere instability in chromosome 1. Five papers were found which were all in English, with total of 29 patients. Forty-three mutations were reported, including 34 missense, 2 deletion, 1 insertion, 6 splice site mutations. Eleven patients had complex heterozygosis mutations. All patients had centromere instability, humoral immune deficiency and facial dysplasia which were mainly ocular hypertelorism and low-set ears. Most patients had language and motor development delay, and a few were combined with mental retardation. Conclusions: ICF syndrome is a rare autosomal recessive primary immunodeficiency with classic clinical triad manifestations. De novo mutation of DNMT3B gene is one of etiologies according to genetic test.

[One year after the implementation of 2015 Tobacco Control Regulation on persons aged 15 years and over tobacco use in Beijing].

Objective: To understand the current status of smoking and smoking cessation in persons aged 15 years and over in Beijing and evaluate the effect of 2015 Beijing Tobacco Control Regulation. Methods: In 2014 and 2016, based on the principles and methodology of the Global Adult Tobacco Survey. A total of 50 communities or townships were selected from 324 communities or townships in Beijing through multistage cluster sampling, and 2 community (village) committees from each community or township were selected with the method of probability proportional to size (PPS). A total of 100 surveillance sites were set, and 100 households were selected from each surveillance site by using simple random sampling. Data were collected through face-to-face interview from the eligible family members aged 15 years and over with the assistance of a tablet computer. Statistical analyses were conducted by using complex sampling analyses module of SPSS 20.0, with weights as a combination of sampling weights, non-response weights and post- stratification weights, for the calculation of current smoking prevalence, daily smoking prevalence, smoking cessation rate, etc. Results: A total of 8 484 and 9 372 valid questionnaires were obtained, respectively, in 2014 and 2016, with the response rate of 86.5% and 96.5%. The current smoking prevalence in persons aged 15 years and over was 23.4% in 2014, and 22.3% in 2016. According to the 6(th) national census data, the current smoking population decreased by 199 000 in Beijing. The proportion of daily smokers declined from 20.7% in 2014 to 19.2% in 2016. The daily number of cigarettes consumed by current smokers increased from 14.6 in 2014 to 15.4 in 2016. The smoking cessation rate was 14.9% in 2014 and 16.8% in 2016. The proportion of current smokers who had at least one smoking cessation attempt in the past 12 months increased from 22.3% in 2014 to 23.2% in 2016, and the proportion of current smokers who planned to quit smoking increased from 11.6% to 15.5%. Among the current smokers who had visited doctors in the past 12 months, the proportion of those having smoking cessation advice was 58.9% in 2014 and 59.2% in 2016. In 2016, among the current smokers who had attempted to quit in the past 12 months, 36.8% were aware of the smoking cessation clinics, and 29.5%were aware of the quitline. Among those who were aware, only 7.7% had actually visited the cessation clinics, and 5.5% had used the quitline. Conclusions: After the implementation of 2015 Beijing Tobacco Control Regulation for 1 year, the current smoking prevalence in persons aged 15 years and over showed decreasing. It is necessary to further prompt the expansion of smoking cessation service to cover more current smokers.

[The role of prophylactic use of pegylated recombinant human granulocyte colony-stimulating factor(PEG-rhG-CSF) in breast cancer receiving adjuvant chemotherapy].

Objectives: To investigate the efficacy and safety of prophylactic use of pegylated recombinant human granulocyte colony-stimulating factor(PEG-rhG-CSF) in breast cancer receiving docetaxel as adjuvant chemotherapy. Methods: A total of 58 patients with breast cancer receiving adjuvant chemotherapy with docetaxel were included between January 2014 to October 2017. Prophylactic use of PEG-rhG-CSF was administered.Patients were further divided into two groups according to the frequency of PEG-rhG-CSF use: frequent use group (≥3 cycles) and non-frequent use group (<3 cycles). Results: There were significant differences in the incidence rates of grade 3/4 neutropenia between the prophylactic group and non-prophylactic group in cycle 1-3(P<0.05). Less febrile neutropenia (FN) was also noted in the prophylactic group compared with the non-prophylactic group in cycle 1 and cycle 3 (P<0.05). Grade 3/4 neutropenia and FN were less in the frequent use of group compared with the non-frequent use group(P<0.001). The most common side effects of PEG-rhG-CSF included fatigue (10.2%), bone joint pain(50.8%), and 2 patients (3.4%) refused further treatment because of bone joint pain. Conclusions: PEG-rhG-CSF should be prophylactically used for preventing neutropenia and febrile neutropenia in breast cancer patients receiving adjuvant chemotherapy with docetaxel regimen.

miR-182 selectively targets HOXA10 in goat endometrial epithelium cells in vitro.

Proper HOXA10 expression was essential for endometrial receptivity what was crucial for successful embryo implantation in mammalian. This study confirmed that miR-182 regulated the expression levels of HOXA10 by binding to its 3' UTR, selectively downregulated HOXA10 in goat endometrial epithelium cells (gEECs) but not stromal cell (gESCs) in vitro. However, HOXA10 and miR-182 both up-expressed in the goat endometrium at gestational day 15 (D15) compared with gestational day 5 (D5), suggesting that there were some other factors regulated the expression of HOXA10 during the development of goat endometrium in vivo. What's more, HOXA10 gene silencing (HOXA10-siRNA) resulted in gEECs apoptosis in vitro, and it regulated the protein levels of oestrogen receptor a (ERa), progesterone receptor B (PRb), insulin-like growth factor 1 receptor (IGF1R), BCL-2, pleiotrophin (PTN), AKT and p-JNK in gEECs. Furthermore, HOXA10 might regulate the protein levels of endometrial receptivity biomarker genes, including vascular endothelial growth factor (VEGF), osteopontin (OPN), cyclooxygenase-2 (COX-2) and prolactin receptor (PRLR) in gEECs. In conclusion, miR-182 targeted HOXA10 selectively in EECs in vitro, and HOXA10 played an important role in maintaining the function of EECs in dairy goats.

Analysis on difference in gastrointestinal hormone levels of patients with the history of diabetes and concurrent nephropathy and study on the role of liraglutide.

OBJECTIVE: To compare the difference in the gastrointestinal hormone levels of the patients with the history of diabetes and concurrent nephropathy and investigate the clinical effect of liraglutide in the treatment of diabetic nephropathy (DN). PATIENTS AND METHODS: 42 cases of patients with DN admitted in our hospital from April 2010-May 2015 were selected and divided into phase I-II group (group A, n = 22) and phase III-IV group (group B, n = 20) according to DN phases and 20 cases of patients with diabetes rather than nephropathy admitted in our hospital during the same period were selected as the control group, all of whom underwent the routine biochemical test and gastrointestinal hormone test, the differences in gastrin (GAS), motilin (MTL) and glucagon (GLC) of DN patients were compared at different phases, the gastric emptying test was carried out on them and the gastric emptying time was recorded. All patients were treated with liraglutide and the changes in fasting blood glucose (FBG), glycosylated hemoglobin (HbAlc), serum creatinine (Cr), blood urea nitrogen (BUN), insulin (FINS) and insulin resistance level (HOMA-IR) were tested before treatment and after 10 weeks' treatment, the changes in the tumor necrosis factor (TNF-α), interleukin -6 (IL-6) and transforming growth factor (TGF-ß1) were determined, and the change in the gastrointestinal hormone levels of patients was recorded after treatment. RESULTS: (1) the GAS, MTL, GLC and gastric emptying time in group B were higher than those in group A and the control group (p < 0.05), and the above indicators in group A were higher (p < 0.05); (2) after 10 weeks' treatment, the gastrointestinal hormone levels in the three groups were reduced and the gastric emptying time was shortened, the difference was statistically significant (p < 0.05) compared with those before treatment, after 10 weeks' treatment, the GAS, MTL, GLC and gastric emptying time in group B were higher than group A and the control group, those in group A were higher than control group (p < 0.05); (3) before treatment, the comparative differences in FBG, HbAlc, FINS and HOMA-IR among the three groups were not statistically significant (p > 0.05), and after 10 weeks' treatment, the differences in FBG, HbAlc and HOMA-IR among three groups were reduced and FINS was increased, the difference in those between before treatment and after treatment was statistically significant (p < 0.05) and the comparative difference among the three groups was not statistically significant (p > 0.05); (4) before treatment, Cr and BUN levels in group A and group B were higher than the control group (p < 0.05), after 10 weeks' treatment, the Cr and BUN levels among three groups were significantly decreased (p < 0.05), Cr and BUN in group A and group B were higher than the control group, cr and BUN levels in group B were higher than group A (p < 0.05); (5) before treatment, the difference by comparing IL-6, TNF-α and TGF-ß1 among three groups were not statistically significant (p > 0.05), after 10 weeks' treatment, the indicators in the three groups were decreased significantly (p < 0.05), but the comparative difference among the three groups were not statistically significant (p > 0.05); (6) the difference by comparing the efficiencies among the three treatment was not statistically significant (p > 0.05). CONCLUSIONS: There are some correlations between the gastrointestinal hormone levels and the degree of renal impairment of DN patients. Good results will be achieved by applying liraglutide in intervention with different phases of DN and DM patients, which cannot only regulate the gastrointestinal hormone levels and lower the blood sugar levels of patients, but can also reduce the insulin resistance and delay the process of renal damage.

Nasal immunization with inhibin DNA vaccine delivered by attenuated Salmonella choleraesuis for improving ovarian responses and fertility in cross-bred buffaloes.

This study was conducted to determine the effect of immunization with inhibin DNA vaccine delivered by attenuated Salmonella choleraesuis on ovarian responses and fertility in cross-bred buffaloes. A total of 134 cross-bred buffaloes were divided into four groups: groups T1 (n = 34), T2 (n = 35) and T3 (n = 31) were nasal immunized twice a day with 10 ml of 1 × 1010  CFU/ml of the C501 (pVAX-asd-IS) vaccine for 5, 3 and 1 day, respectively. Group C (n = 34) was nasal immunized with 10 ml PBS for 5 days. All animals were immunized twice with an interval of 14 days and administered with 200 µg of a GnRH analogue on day 28, 0.5 mg PGF2α on day 35 and 200 µg of the same GnRH analogue on day 37. TAI was performed at 18 and 24 hr after the second GnRH treatment. Fourteen days after primary immunization, C501 (pVAX-asd-IS) elicited significant immune responses, and anti-inhibin IgG antibody titres in group T1 were significantly higher (p < .01) than groups T3 and C. After the second GnRH treatment, the growth speed of the dominant follicles in group T1 was significantly faster (p < .05) than groups T3 and C. The number and diameter of large follicles (≥10 mm) as well as ovulatory follicles in group T1 were the greatest in all groups, resulting in a greater conception rate in buffaloes with positive anti-inhibin antibodies. These results demonstrate that immunization with the C501 (pVAX-asd-IS) vaccine, coupled with the Ovsynch protocol, could be used as an alternative approach to improve reproductive performance in cross-bred buffaloes.

Soluble and membrane-bound interleukin (IL)-15 Rα/IL-15 complexes mediate proliferation of high-avidity central memory CD8+ T cells for adoptive immunotherapy of cancer and infections.

The lack of persistence of infused T cells is a principal limitation of adoptive immunotherapy in man. Interleukin (IL)-15 can sustain memory T cell expansion when presented in complex with IL-15Rα (15Rα/15). We developed a novel in-vitro system for generation of stable 15Rα/15 complexes. Immunologically quantifiable amounts of IL-15 were obtained when both IL-15Rα and IL-15 genes were co-transduced in NIH 3T3 fibroblast-based artificial antigen-presenting cells expressing human leucocyte antigen (HLA) A:0201, ß2 microglobulin, CD80, CD58 and CD54 [A2-artificial antigen presenting cell (AAPC)] and a murine pro-B cell line (Baf-3) (A2-AAPC15Rα/15 and Baf-315Rα/15 ). Transduction of cells with IL-15 alone resulted in only transient expression of IL-15, with minimal amounts of immunologically detectable IL-15. In comparison, cells transduced with IL-15Rα alone (A2-AAPCRα ) demonstrated stable expression of IL-15Rα; however, when loaded with soluble IL-15 (sIL-15), these cells sequestered 15Rα/15 intracellularly and also demonstrated minimal amounts of IL-15. Human T cells stimulated in vitro against a viral antigen (CMVpp65) in the presence of 15Rα/15 generated superior yields of high-avidity CMVpp65 epitope-specific T cells [cytomegalovirus-cytotoxic T lymphocytes (CMV-CTLs)] responding to ≤ 10- 13 M peptide concentrations, and lysing targets cells at lower effector : target ratios (1 : 10 and 1 : 100), where sIL-15, sIL-2 or sIL-7 CMV-CTLs demonstrated minimal or no activity. Both soluble and surface presented 15Rα/15, but not sIL-15, sustained in-vitro expansion of CD62L+ and CCR7+ central memory phenotype CMV-CTLs (TCM ). 15Rα/15 complexes represent a potent adjuvant for augmenting the efficacy of adoptive immunotherapy. Such cell-bound or soluble 15Rα/15 complexes could be developed for use in combination immunotherapy approaches.

[Efficacy and safety analysis of surgical bypass and endovascular management in the treatment of 116 Takayasu arteritis].

OBJECTIVE: To analyze the efficacy and safety of surgery and endovascular management in treating Takayasu arteritis. METHODS: The data of 116 patients (24 males and 92 females; mean age (32±12) years) with Takayasu arteritis and underwent surgery or endovascular therapy was retrospective analyzed. According to the two different surgical procedures, the patients were divided into two groups: open repair group and endovascular repair group. One hundred and fifty-four surgical procedures were done including 69 cases of open repair and 85 cases of endovascular repair. A total of 211 arterial lesions were revascularized (open repair 114; endovascular repair 97). RESULTS: Among the 154 surgical procedures, 11(7.1%) presented a complication during perioperative period including 6(8.7%) of open repair and 5(5.9%) of endovascular repair. After a median follow-up of 38.5(0.5-142.0) months, three(4.3%) cases of stroke and death were observed in open repair group, two(2.3%) cases of stroke and 4(4.7%) cases of death were observed in endovascular repair group. At 1, 3, 5 and 10 years of follow-up, primary patency rate of open repair and endovascular repair were 95.0% and 89.3%, 84.3% and 69.8%, 73.3% and 56.3%, 53.4% and 48.1%, respectively; Primary assisted patency rate were 100% and 97.5%, 90.4% and 78.2%, 79.1% and 72.8%, 60.7% and 54.0%, respectively; Secondary patency rate were 100% and 98.8%, 95.6% and 92.7%, 85.8% and 78.1%, 74.8% and 58.0%, respectively. Cumulative survival rate were 97.0% and 100%, 97.0% and 97.6%, 97.0% and 90.6%, 91.3% and 84.5%, respectively (χ(2)=0.182, P=0.669). CONCLUSIONS: Both of the surgical revascularization and endovascular management are safe and effective in the treatment of Takayasu arteritis. Although long-term patency of endovascular therapy is low, it can be performed repeatedly and can be used as a preferred approach in treating a short stenosis. Surgical repair shows excellent long-term durability, it seems to be more suitable for complex lesions and failure cases of endovascular management.

Oxidative stress inhibits growth and induces apoptotic cell death in human U251 glioma cells via the caspase-3-dependent pathway.

OBJECTIVE: To investigate the possible pathway involved in hydrogen peroxide (H2O2) induced apoptosis in U251 glioma cells. MATERIALS AND METHODS: The cultured U251 glioma cells were used in this study. The cells were divided into three groups: control group (untreated glioma cells), H2O2 group (treated with 100, 300 and 500 µM H2O2) and CI group (treated with 300 µM H2O2 and 15 µM caspase inhibitor, CI). The cellular viability was determined by MTT [3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide] assay. A flow cytometer was used for measuring the cell cycles. The mode of cell death was assessed by Annexin-V/PI-flow cytometry analysis. Fluorescence dihydroethidium (DHE) method was conducted to detect the oxygen species (O2-). Western blot analysis was performed to confirm the pro-caspase-3, caspase-3 and PARP (poly-ADP-ribose polymerase) protein expression. RESULTS: The oxidative stress to U251 glioma cells exhibited in a dose-dependent manner with H2O2 concentrations increasing. The cell viability was considerably decreased and apoptosis occurred in H2O2 treated cells. A G1 cell cycle arrest and O2- level increase were found in H2O2 group. Western blot analysis showed a decrease of pro-caspase-3 protein level and an increase of caspase-3 and PARP level in 300 µM H2O2 treated cells. The H2O2 induced apoptosis depicted above was significantly restrained by CI. CONCLUSIONS: Oxidative stress inhibits growth and induces apoptotic cell death in human U251 glioma cells via the caspase-3-dependent pathway. Mitochondrial pathway might involve in this signaling conduction. These findings are favorable for understanding the mechanisms of oxidative stress-induced apoptosis in U251 glioma cells.

Unravelling the multifaceted roles of Atg proteins to improve cancer therapy.

Autophagy follows a lysosomal degradation pathway in which a cell digests its own components. It is highly regulated by a limited number of autophagy-related genes (Atg) and the proteins they encode, that are crucial for cells to undergo the process via modulating autophagsome formation. Recently, accumulating evidence has revealed the core molecular machinery of autophagy; however, intricate relationships between autophagy and cancer remain an enigma. Several studies have shown that Atgs can play an important role in carcinogenesis, by which Atgs may modulate a series of oncogenic and tumour suppressive pathways, implicating microRNA (miRNA) involvement. In this review, we will present the key role of Atgs in deciding the fate of cancer cells, discuss some representative Atgs and their proteins such as ULK, Beclin-1, and Atg8/LC3-Atg4, which can also be regulated by miRNAs. Thus, Atgs can be considered to be targets for cancer treatment, which may illuminate the future of cancer therapy.

Complete eradication of hepatomas using an oncolytic adenovirus containing AFP promoter controlling E1A and an E1B deletion to drive IL-24 expression.

Interleukin (IL)-24, a promising therapeutic gene, has been widely used for Cancer Targeting Gene-Viro-Therapy (CTGVT). In this study, IL-24 was inserted into an oncolytic adenovirus in which the E1A gene is driven by an enhanced, short α-fetoprotein (AFP) promoter and the E1B gene is completely deleted to form Ad.enAFP-E1A-ΔE1B-IL-24. This construct has a potent antitumor effect on liver cancer cell lines in vitro, but little or no effect on normal cell lines, such as L-02 and QSG-7701. In vivo, the complete elimination of Huh-7 liver cancer in nude mice with Ad.enAFP-E1A-ΔE1B-IL-24 intratumor injection was observed. The design of Ad.enAFP-E1A-ΔE1B-IL-24 and its potent antitumor effect on liver cancer have not been published previously. The mechanism of the potent antitumor effect of Ad.enAFP-E1A-ΔE1B-IL-24 is due to the upregulation of GADD34 and intrinsic and extrinsic apoptotic signaling.

Cancer targeting Gene-Viro-Therapy of liver carcinoma by dual-regulated oncolytic adenovirus armed with TRAIL gene.

Liver cancer is a common and aggressive malignancy, but available treatment approaches remain suboptimal. Cancer targeting Gene-Viro-Therapy (CTGVT) has shown excellent anti-tumor effects in a preclinical study. CTGVT takes advantage of both gene therapy and virotherapy by incorporating an anti-tumor gene into an oncolytic virus vector. Potent anti-tumor activity is achieved by virus replication and exogenous expression of the anti-tumor gene. A dual-regulated oncolytic adenoviral vector designated Ad·AFP·E1A·E1B (Δ55) (Ad·AFP·D55 for short thereafter) was constructed by replacing the native viral E1A promoter with the simian virus 40 enhancer/α-fetoprotein (AFP) composite promoter (AFPep) based on an E1B-55K-deleted construct, ZD55. Ad·AFP·D55 showed specific replication and cytotoxicity in AFP-positive hepatoma cells. It also showed enhanced safety in normal cells when compared with the mono-regulated vector ZD55. To improve the anti-hepatoma activities of the virus, the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene was introduced into Ad·AFP·D55. Ad·AFP·D55-TRAIL exhibited remarkable anti-tumor activities in vitro and in vivo. Treatment with Ad·AFP·D55-TRAIL can induce both autophagy owing to the Ad·AFP·D55 vector and caspase-dependent apoptosis owing to the TRAIL protein. Therefore, Ad·AFP·D55-TRAIL could be a potential anti-hepatoma agent with anti-tumor activities due to AFP-specific replication and TRAIL-induced apoptosis.

CD2 signaling in T cells involves tyrosine phosphorylation and activation of the Tec family kinase, EMT/ITK/TSK.

Ligation of the CD2 cell surface glycoprotein expressed on T lymphocytes and NK cells induces protein tyrosine phosphorylation and activation of the Src kinases, LCK and FYN. We show here that in Jurkat T leukemia cells and in peripheral blood T cells, CD2 stimulation also leads to tyrosine phosphorylation and activation of the Tec family kinase, EMT/ITK/TSK. Activation of EMT by CD2 was induced by mitogenic pairs of CD2 mAb, certain single CD2 mAb followed by secondary antibody cross-linking, and CD58-bearing sheep red blood cells. With the use of different Jurkat cell mutants it was demonstrated that CD2-mediated activation of EMT required expression of LCK, but not require surface expression of the CD3 zeta chain. Receptor-mediated activation of LCK does not in itself lead to activation of this Tec kinase since induction of LCK by ligation of CD4 or CD5 did not result in activation of EMT. The activation of EMT during CD2 signaling suggests an important role for this kinase in CD2 co-stimulation of T cell responses.

CD28-mediated cytotoxicity by the human leukemic NK cell line YT involves tyrosine phosphorylation, activation of phosphatidylinositol 3-kinase, and protein kinase C.

The human leukemic cell line YT displays spontaneous cytotoxicity against CD80+ and/or CD86+ and ICAM-1+ target cells. In this work, we report that CD28-mediated cytotoxicity of YT involves tyrosine phosphorylation and activation of phosphatidylinositol (PI) 3-kinase, the Tec kinase Itk/Emt, and protein kinase C (PKC). YT mediates lysis of CD80+/CD86+ B lymphoblastoid cell lines and the murine mastocytoma p815 transfected with CD80 or CD86. The lysis was inhibited by two different Pi 3-kinase inhibitors, wortmannin and LY294002. The PKC inhibitors calphostin C and bisindolylmaleimide GF109203X also abolished YT-mediated cytotoxicity. Furthermore, exocytosis of cytolytic effector molecules was also inhibited by PI 3-kinase inhibitors and PKC inhibitors. PMA together with Ionomycin did not induce granule exocytosis or cytotoxicity by YT cells. Treatment of YT cells with PMA for up to 20 h, which depleted PMA-responsive PKC isoforms, had no effect on the CD28-mediated cytotoxicity. This cytotoxicity displayed by PMA-treated YT cells, however, could still be inhibited by Pi 3-kinase inhibitors and PKC inhibitors. Taken together, these results are consistent with a model in which activation of CD28 and LFA-1 induces tyrosine phosphorylation of the CD28 cytoplasmic domain, recruitment and activation of PI 3-kinase, as well as the Tec kinase Itk/Emt, and the activation of PMA-nonresponsive PKC isoenzymes. Activation of PI 3-kinase and PMA-nonresponsive PKC isoenzymes is shown to be involved directly in cytolytic granule release by YT cells.

[Treatment of intestinal metaplasia and atypical hyperplasia of gastric mucosa with xiao wei yan powder].

UNLABELLED: 138 cases of intestinal metaplasia (IM) and 104 cases of atypical hyperplasia (AH) of the gastric mucosa of chronic gastritis treated with Xiao Wei Yan Powder (XWYP) were reported. The diagnoses were based on the pathological examination of gastric antrum biopsy specimens. The cases were randomly divided into treated group and control group. The XWYP contained Smilax glabrae, Hedyotis diffusae, Taraxacum mongolicum, Caesalpinia sappan, Paeonia alba, Cyperus rotundus, Bletilla striata, Glycyrrhiza uralensis etc., and was prepared in powder form, taken orally 5-7g tid. After 2-4 months of administration, gastroscopic and pathological examinations were repeated. RESULTS: In treated group, the total effective rate of IM was 91.3% and that of the AH was 92.16%, while in control group, they were 21.3% and 14.46% respectively (P < 0.01). It denoated that XWYP had marked therapeutic effects for IM and AH. The animal experiments revealed no toxic effect, so safety guarantee was provided for its clinical application.