[Analysis of risk factors of short-term prognosis in patients with severe Budd-Chiari syndrome].

Objective: To explore the risk factors of short-term prognosis of severe Budd-Chiari syndrome (BCS) patients,established and verified the nomogram prediction model for these BCS patients and evaluated its clinical application value. Methods: This study is a retrospective cohort study. The clinical data of 171 patients with severe BCS diagnosed were retrospectively analyzed in the Department of Hepatopancreatobiliary Surgery First Affiliated Hospital of Zhengzhou University from January 2018 to December 2023. There were 105 males and 66 females, aged (52.1±12.8) years (range: 18 to 79 years). The patients were divided into two groups based on whether they died within 28 days: the death group (n=38) and the survival group (n=133). The risk factors for short-term death of patients were analyzed,and independent risk factors were screened by univariate and multivariate analysis. Furthermore,these factors were used to establish the nomogram prediction model. The area under the curve(AUC),the Bootstrap Resampling,the Hosmer-Lemeshow test and the Decision Curve Analysis(DCA) were used to verify the model's differentiation,internal verification,calibration degree and clinical effectiveness,respectively. Results: Univariate and multivariate Logistics regression analysis showed that the history of hepatic encephalopathy,white blood cell,glomerular filtration rate and prothrombin time were independent risk factors (P<0.05). The above factors were used to successfully establish the prediction model with 0.908 of AUC and 0.895 of the internal verification of AUC,indicating that the predictive model was valuable. The 0.663 P-values in the Hosmer-Lemeshow test indicated the high calibration degree of the model. The clinical effectiveness of the model was proved by the 18% clinical benefit population using the DCA curve with the 17% probability threshold. Conclusions: The independent risk factors are the history of hepatic encephalopathy,white blood cell,glomerular filtration rate and prothrombin time. An adequate basis was acquired by establishing a nomogram prediction model of the short-term prognosis of severe BCS,which was helpful for early clinical screening and identification of high-risk patients with severe BCS who could die in the short term and timely providing timely intervention measures for improving the prognosis.

[Emphasizing the standard application of toric intraocular lenses].

In the era of refractive cataract surgery, corneal astigmatism is a crucial factor that affects visual quality after cataract surgery. Toric intraocular lenses (IOLs) have a wide correction range, good surgical predictability, minimal damage to the ocular surface, and stable long-term effects, which can significantly improve postoperative visual quality and patient satisfaction. However, there are still several challenges in the clinical application of toric IOLs, including the awareness of astigmatism correction, the accuracy of preoperative biometry, the widespread adoption of the concept of total corneal astigmatism, the management of surgical planning and details, and the control of postoperative residual astigmatism. This article aims to address these issues to further standardize the clinical application of toric IOLs and help improve the overall level of refractive cataract surgery in China.

[The risk of incident gastric cancer for populations with different precancerous gastric lesions: a prospective follow-up study].

Objective: To provide evidence for optimizing the screening strategy for gastric cancer (GC), we evaluated the risk of incident GC for individuals with different precancerous gastric lesions in a prospective cohort study. Methods: Based on the National Upper Gastrointestinal Cancer Early Detection Program launched in Linqu, Shandong, a high-risk area of gastric cancer in China, we included a total of 14 087 subjects diagnosed with different gastric lesions stages by endoscopic screening from 2012 to 2018. Study subjects were prospectively followed up until December 31, 2019. The incidence of GC during the follow-up was ascertained by repeated endoscopic examinations, cancer, death registry reports, and active follow-up of study subjects and was confirmed by reviewing medical records extracted from the hospital information management system. The Poisson regression model was applied to calculate the relative risk (RR) and 95%CI for GC occurrence among subjects with different gastric lesions. Results: Among 14 087 subjects with different gastric lesions as determined by their first endoscopic examination in 2012-2018, 7 608 (54.00%) had a global diagnosis of superficial gastritis (SG), 2 848 (20.22%) had chronic atrophic gastritis (CAG), 3 103 (22.03%) had intestinal metaplasia (IM), and 520 (3.69%) had low-grade intestinal neoplasia (LGIN). During the follow-up, 109 subjects were diagnosed with GC, including 63 with high-grade intestinal neoplasia (HGIN) and 46 with invasive GC. Compared to subjects having normal gastric mucosa or SG, those with CAG (RR=3.85, 95%CI: 2.04-7.28), IM (RR=5.18, 95%CI: 2.79-9.60), and LGIN (RR=19.08, 95%CI: 9.97-36.53) had significantly increased risk of progression to GC. Individuals with these gastric lesions had an elevated risk of developing HGIN and invasive GC. For subjects with LGIN, the RR was 22.96 (95%CI: 9.71-54.27) for developing HGIN and 14.64 (95%CI: 5.37-39.93) for developing invasive GC. Subgroup analyses found that all age group subjects with LGIN diagnosed during the initial endoscopic examination had a significantly increased risk of developing the GC. Conclusions: Our large-scale prospective study on a high-risk area of GC showed that most residents aged 40-69 years had gastric lesions of different stages. Subjects with more advanced gastric lesions had a significantly increased risk of progression to GC.

[The effect of cervical lymph node metastasis density on prognosis of hypopharyngeal carcinoma].

Objective: To investigate the relationship between the cervical lymph node density (LND) and the prognosis of hypopharyngeal carcinoma. Methods: The clinical and pathological data of 241 patients with hypopharyngeal carcinoma who underwent surgery in Shandong Provincial ENT Hospital from January 2014 to December 2017 were retrospectively analyzed, including 229 males and 12 females, aged 37-81 years. The LND was calculated, i.e. the ratio of the number of lymph nodes with metastasis to the total number of lymph nodes removed. The patients were divided into low LND group and high LND group based on the cutoff value of LND determined by receiver operating characteristic curve (ROC curve). The univariate and multivariate analyses of the disease-free survival (DFS) and the overall survival (OS) were performed in two groups. Results: With the cutoff value of 0.068, 165 patients were in the low LND group (<0.068) and 76 patients in the high LND group (≥0.068). T stage, N stage, maximum lymph node diameter, extracellular invasion of lymph node, and postpharyngeal lymph node metastasis were associated with LND (statistical values were -3.15, -6.82, 23.37, 20.44, and 30.18, respectively, all P values were<0.05). The univariate analysis showed that age, T stage, N stage, maximum diameter of cervical lymph nodes, extracapsular invasion, retropharyngeal lymph node metastasis and LND were the main factors affecting the patients' DFS (χ2=9.31, 7.30, 20.09, 15.30, 9.04, 19.44, 50.27, all P values<0.05) and OS (χ2 were 5.02, 12.94, 18.28, 15.91, 7.95, 16.88, 49.45, all P values<0.05). Multivariate analysis showed that patients with age≤60 years old and LND≥0.068 had reduced DFS [HR values were 0.61 (95%CI 0.43-0.88) and 2.23 (95%CI 1.44-3.45), both P values<0.05]; patients with advanced T stage and LND≥0.068 had reduced OS [HR values were 1.73 (95%CI 1.02-2.93) and 2.39 (95%CI 1.51-3.80), both P values<0.05]. Conclusion: LND is a prognostic factor for patients with hypopharyngeal carcinoma after surgery, with worse prognosis in patients with LND≥0.068.

Beneficial effect of melatonin administration on the function of frozen-thawed rat testicular grafts.

BACKGROUND: Spermatogonia in testis is sensitive to the cytotoxicity of chemotherapy agents. Cryopreservation of testicular tissue may offer fertility restoration in young male cancer survivors. OBJECTIVE: To investigate the effect of melatonin on the survival of testicular grafts following cryopreservation and transplantation. MATERIALS AND METHODS: Wister rats were randomly allocated into three groups: control group; saline group (cryopreservation + autograft + saline); and melatonin group (cryopreservation + autograft + melatonin). Malondialdehyde (MDA) content, glutathione peroxidase (GPx) activity and superoxide dismutase (SOD) activity were assessed on day 7 after autograft transplantation. At day 30, graft recovery, spermatogonia per round tubule, and serum testosterone concentration in grafts were measured. RESULTS: Melatonin significantly diminished MDA content, enhanced GPx and SOD activities. Furthermore, the recovery rate, number of spermatogonia per round tubule, and serum testosterone concentration in melatonin group was markedly higher than the saline group. CONCLUSION: Melatonin administration at 20 mg/kg is effective in improving the function of frozen and thawed rat testicular graft. The protective role of melatonin can be attributed partly to the enhanced ROS scavenging and antioxidant enzyme activities. doi.org/10.54680/fr22310110612.

[Analysis of early recurrence within 6 months in patients of esophageal squamous cell carcinoma after neoadjuvant therapy followed by surgery].

Objectives: To examine the prognosis factors of recurrence of esophageal carcinoma within 6 months after neoadjuvant therapy followd by surgery. Methods: The clinical data of 187 patients with esophageal squamous cell carcinoma who underwent neoadjuvant therapy followed by curative esophagectomy between January 2018 and April 2020 at Department of Thoracic Surgery, Shanghai Chest Hospital were analyzed retrospectively. There were 160 males and 27 females, aging (63.0±7.1) years (range:43 to 76 years). The t test, χ2 test and rank-sum test were used for univariate analysis of the prognosis factors for recurrence within 6 months postoperative, while the Logistic regression was used for multivariate analysis. Results: There were 30 patients (16.0%) developed recurrence within 6 months after operation, including local recurrence in 1 case, regional recurrence in 11 cases, hematogenous recurrence in 13 cases, and combined recurrence in 5 cases. Univariate analysis suggested that there was a significant difference in T staging of tumor before neoadjuvant therapy (cT), tumor regression grade, circumferential resection margin, pathological T stage (ypT) and pathological N stage (ypN) between the recurrence patients and non-recurrence patients (all P<0.05). Logistic regression analysis suggested that the cT3-4 (OR=2.701, 95%CI: 1.161 to 6.329, P=0.021) and ypN(+)(OR=1.654, 95%CI: 1.045 to 2.591, P=0.032) were the independent prognosis factors for recurrence within 6 months. Conclusion: The combination of neoadjuvant therapy and surgery is not effective in reducing early postoperative recurrence in patients who have invaded the epineurium before treatment, and still have positive lymph nodes after neoadjuvant therapy.

[Pathological characteristics and molecular diagnosis of non-tuberculosis Mycobacterium lung disease].

Objective: To investigate the clinicopathological features of non-tuberculosis mycobacterial lung disease and the role of molecular pathology in diagnosis. Methods: Forty-five formalin-fixed, paraffin embedded (FFPE) specimens were collected from the Department of Pathology, Beijing Chest Hospital from February 2016 to August 2019. The clinical, imaging and histopathologic features, bacteriologic data and morphologic characteristics of acid fast bacilli (AFB) were analyzed retrospectively. Specific gene sequence IS6110 of Mycobacterium tuberculosis (MTB) was detected by fluorescence PCR. Identification of Mycobacteria was by melting curve method. Fifty cases of pulmonary tuberculosis were selected in the same period as control. Results: The NTM lung cases included 18 cases (40.0%, 18/45) of M. intracellulare, eight cases (17.8%, 8/45) of M. xenopi, six cases (13.3%, 6/45) of M. avium, six cases (13.3%, 6/45) of M. kansasii, six cases (13.3%, 6/45) of M. chelonae and one case (2.2%, 1/45) of M. simiae. Histopathologically, there were necrotizing granulomas in 34 cases (75.6%, 34/45), non-necrotizing granuloma in one case (2.2%, 1/45) and non-granulomatous lesions in 10 cases (22.2%, 10/45). The necrosis was pink necrosis, basophilic necrosis rich in nuclear fragments and suppurative necrosis. Pulmonary TB showed more pink necrosis and basophilic necrosis, the difference was statistically significant (χ(2)=10.270, P=0.001; χ(2)=7.449, P=0.006). Seventeen cases (37.8%, 17/45) of NTM lung disease showed giant multinucleated giant cells, which were significantly different from those in pulmonary tuberculosis group (χ(2)=13.446, P<0.01). The number and morphology of AFB were also different. More AFB were found in M. intracellular cases and significant AFB were easily seen in M. kansasii infection. Conclusions: M. tuberculosis and NTM cannot be reliably differentiated by histologic features or by AFB morphology. Molecular assays are important to distinguish tuberculosis from NTM lung disease.

[Surgical key points of precision functional sphincter-preserving surgery].

The principle of sphincter-preserving surgery is to preserve the anal sphincter function under the premise of radical resection. Due to low position of rectal tumor, conventional laparoscopic surgery has difficulties in operating in the deep and narrow pelvis, which may lead to inaccurate tissue dissociation, imprecise positioning of tumor edge, excessive stretch of the anal sphincter complex, and excessive removal of distal rectal mucosa. Moreover, pain from abdominal auxiliary incision has an unavoidable side effect for postoperative recovery. With the help of the Liu's transanal microsurgery system, precision functional sphincter-preserving surgery (PPS) can be successfully performed. PPS tries to preserve left colonic artery and pelvic autonomic nerve in the transabdominal operation. In the part of transanal surgery, measurement, localization and resection of the lower edge of the tumor are conducted under a clear and open visual field with the transparent screw anal dilator. After the rectum is cut off, the specimen is taken out through the anus to avoid abdominal incision. Inserting the intestinal supporter to support the bowel stump, full thickness of bowel stump is then sutured with anal canal by vertical mattress suture. Special transanal tube is placed afterwards without routine prophylactic stoma. PPS can achieve precise tumor resection and sphincter preservation simultaneously.

Low-expression of lncRNA-ANCR promotes tibial fracture healing via targeting RUNX2.

OBJECTIVE: To explore the role of long non-coding ribonucleic acid-anti-differentiation non-coding ribonucleic acid (lncRNA-ANCR) in tibial fracture healing in rabbits by regulating the runt-related transcription factor 2 (RUNX2) expression. MATERIALS AND METHODS: A total of 60 healthy adult rabbits were evenly divided into Control group (n=20), Fracture group (n=20), and Lnc group (n=20). Then, RUNX2 transfection, Real Time Polymerase Chain Reaction (PCR) assay and relevant instruments were carried out and used to determine the differences in dry weight, bone mineral density, bone mechanical strength, and RUNX2 expression in tibiae among three groups of rabbits. RESULTS: Comparison of the bone mineral density in rabbit tibiae among the three groups showed that the bone mineral density was significantly lower in Fracture group than that in Control group (p<0.05), and it was slightly higher in Lnc group than in Fracture group (p<0.05). The dry weight of the full-length tibiae in Fracture group was significantly decreased compared with that in Control group (p<0.05), and Lnc group had an increased dry weight of tibiae in comparison with Fracture group (p<0.05). The maximum load, flexural strength, elastic stress, elastic strain, elastic modulus, maximum stress, and maximum strain in Fracture group were lower than those in both Control group and Lnc group (p<0.05). Compared with those in Fracture group, the amount of new collagen was overtly increased in Lnc group, and that of mature collagen was decreased (p<0.05). The relative expression level of RUNX2 in tibial bone tissues was evidently lower in Fracture group than that in Control group (p<0.05), and it was markedly higher in Lnc group than that in Fracture group (p<0.05). CONCLUSIONS: Down-regulating lncRNA-ANCR activates and triggers the expression of RUNX2 that facilitates the growth and metabolism of bone tissues to play an important role in the repair of bone tissues and promote the healing of the tibial fracture.

[A survey on tyrosine kinase inhibitor treatment in patients with chronic myeloid leukemia in China: from patients'perspective].

OBJECTIVES: To assess tyrosine kinase inhibitor (TKI) treatment status in patients with chronic myeloid leukemia (CML) in China and analyze the response-associated factors. METHODS: From May to November in 2014, anonymous questionnaires were distributed to adult CML patients who were receiving TKI treatment all over China. RESULTS: 1 038 questionnaires were collected, 949 questionnaires were evaluable. Of the 949 evaluable respondents, 549 (58%) were male with the median age of 41 years (range, 18 to 88 years). 623 (66%) respondents lived in an urban area and 449 (47%) had an education level ≥ a bachelor degree. 888 (94%) respondents were in the chronic phase at diagnosis, and 690 (78%) of them started TKI treatment within one year after diagnosis. 794 (84%) respondents were on imatinib, 768 (81%) on the branded. With a median TKI treatment duration of 3 years (range, <1 to 13 years), 708 of 834 (85%) evaluable respondents achieved Ph- negative (i.e. complete cytogenetic response, CCyR), and 497 of 859 (46% ) BCR- ABL negative (i.e. complete molecular response, CMR). Multivariate analyses showed that female (OR=1.8, 95% CI 1.1-2.8,P=0.019 andOR=1.5, 95% CI 1.1-2.0,P=0.015), TKI treatment duration >3 years (OR=4.1, 95% CI 2.6- 6.5,P<0.001 andOR=3.7, 95% CI 2.7- 5.1,P< 0.001) and imatinib taken (OR=2.1, 95% CI 1.2-3.7,P=0.007 andOR=3.3, 95% CI 2.1-5.1,P<0.001) were factors affecting achieving both CCyR and CMR. In addition, higher education level (OR=2.0, 95% CI 1.3- 3.1,P=0.003), starting TKI treatment <1 year (OR=2.4, 95% CI 1.5- 3.8,P<0.001) and branded drugs received (OR=2.4, 95% CI 1.4- 4.0,P=0.001) were factors affecting achieving a CCyR. In 884 respondents, 534 (62%) reported " heavy financial burden" as the biggest treatment impediment, only 152 (17%) reported " poor quality of life related to adverse effects of TKI". CONCLUSIONS: The survey showed that majority of the Chinese CML patients received imatinib as a TKI therapy, and most of the patients achieved satisfied responses by TKI. Financial burden became the major obstacle during TKI treatment.

Clinical Outcomes of Patients with Severe Hepatic Hereditary Hemorrhagic Telangiectasia After Banding of the Hepatic Artery and Banding/Ligation of Branches of the Hepatic Artery.

OBJECTIVE/BACKGROUND: To evaluate the effectiveness of double banding/ligation of hepatic arteries in treating patients with hepatic hereditary hemorrhagic telangiectasia (HHHT). METHODS: From January 2004 to December 2013, 35 patients were diagnosed with HHHT, among whom 11 woman and two men with a mean ± SD age of 44 ± 9 years were treated by double hepatic artery banding/ligation for cardiac insufficiency and/or portal hypertension. The outcomes were evaluated prospectively by measuring clinical manifestations, imaging features, liver and cardiac function, pulmonary arterial systolic pressure, and post-operative complications. Quality of life was evaluated with the Short Form Health Survey questionnaire. RESULTS: For each patient, the common hepatic artery and one branch of the left and/or right hepatic artery were banded, and other significantly dilated hepatic artery branches were ligated. No patient died after surgery. Clinical symptoms were improved in all patients, although ischemic cholangitis was observed in two patients and treated conservatively. Cardiac function, classified per the New York Heart Association (NYHA) cardiac functional grading, improved (NYHA III-IV vs. NYHA I-II); pulmonary arterial systolic pressure significantly decreased in all patients (48 ± 8 mmHg vs. 24 ± 4 mmHg; P < .001) and remained in the normal range (26 ± 3 mmHg) at the end of follow up. The levels of γ-glutamyl transpeptidase and alkaline phosphatase decreased in 11 patients (144 ± 94 U/L vs. 71 ± 34 U/L; P = .003) and 10 patients (207 ± 71 U/L vs. 105 ± 32 U/L; P = .001), respectively. Patients were followed up for 50 ± 28 months (range 6-113 months); one death resulted from causes unrelated to surgery and all dimensions of quality of life improved in all surviving patients. CONCLUSIONS: This study helps to establish double hepatic artery banding/ligation as an effective therapy for selected patients with HHHT.

Changes in tau phosphorylation levels in the hippocampus and frontal cortex following chronic stress

Studies have indicated that early-life or early-onset depression is associated with a 2- to 4-fold increased risk of developing Alzheimers disease (AD). In AD, aggregation of an abnormally phosphorylated form of the tau protein may be a key pathological event. Tau is known to play a major role in promoting microtubule assembly and stabilization, and in maintaining the normal morphology of neurons. Several studies have reported that stress may induce tau phosphorylation. The main aim of the present study was to investigate possible alterations in the tau protein in the hippocampus and frontal cortex of 32 male Sprague-Dawley rats exposed to chronic unpredictable mild stress (CUMS) and then re-exposed to CUMS to mimic depression and the recurrence of depression, respectively, in humans. We evaluated the effects of CUMS, fluoxetine, and CUMS re-exposure on tau and phospho-tau. Our results showed that a single exposure to CUMS caused a significant reduction in sucrose preference, indicating a state of anhedonia. The change in behavior was accompanied by specific alterations in phospho-tau protein levels, but fluoxetine treatment reversed the CUMS-induced impairments. Moreover, changes in sucrose preference and phospho-tau were more pronounced in rats re-exposed to CUMS than in those subjected to a single exposure. Our results suggest that changes in tau phosphorylation may contribute to the link between depression and AD.

Human endogenous retrovirus W family envelope gene activates the small conductance Ca2+-activated K+ channel in human neuroblastoma cells through CREB.

Numerous studies have shown that human endogenous retrovirus W family (HERV-W) envelope gene (env) is related to various diseases but the underlying mechanism has remained poorly understood. Our previous study showed that there was abnormal expression of HERV-W env in sera of patients with schizophrenia. In this paper, we reported that overexpression of the HERV-W env elevated the levels of small conductance Ca(2+)-activated K(+) channel protein 3 (SK3) in human neuroblastoma cells. Using a luciferase reporter system and RNA interference method, we found that functional cAMP response element site was required for the expression of SK3 triggered by HERV-W env. In addition, it was also found that the SK3 channel was activated by HERV-W env. Further study indicated that cAMP response element-binding protein (CREB) was required for the activation of the SK3 channel. Thus, a novel signaling mechanism of how HERV-W env influences neuronal activity and contributes to mental illnesses such as schizophrenia was proposed.

Ceramide accumulation and up-regulation of proinflammatory interleukin-1ß exemplify lipotoxicity to mediate declines of reproductive efficacy of broiler hens.

The study was conducted to delineate fundamental mechanisms that initiate the deleterious effect of fuel overloading on reproductive efficacy of broiler breeder hens. Sixty hens at age 26 wk were fed recommended amounts of feed (160 g/d per hen) or allowed voluntary feeding (approximately 30% more than restriction). At age 35 and 50 wk, hens were sampled for further analyzes. Voluntary feeding resulted in poor egg production, high rate of mortality, and abnormal ovarian structure (mainly overt hierarchical follicle atresia at age 35 wk and ovarian involution at age 50 wk). In contrast to feed-restricted hens, voluntary feeding also induced metabolic dysregulations that comprised enhanced adiposity; hepatic triacylglycerol accumulation; and elevated concentrations of plasma glucose, NEFAs, very low density lipoprotein, triacylglycerol, phospholipids, and sphingomyelin (P < 0.05). Furthermore, hepatic and circulating ceramide and sphingomyelin accumulation, and up-regulation of proinflammatory IL-1ß expression in liver and adipose tissues (P < 0.05) systemically manifested the development of lipotoxicity in feed-satiated hens. Lipotoxicity leading to impaired ovarian dysfunctions, including follicle atresia, ovarian regression, and a decline of circulating estradiol levels (P < 0.05) in feed-satiated hens, was further exemplified by ceramide accumulation and up-regulation of IL-1ß, serine palmitoyltransferase, and sphingomyelinase transcript abundance, but suppressed protein kinase Akt activation (P < 0.1 to 0.05) within the hierarchical follicles. This study provides the first in vivo evidence of the actions of ceramide and IL-1ß in mediating overfeeding-induced follicle atresia and progression of ovarian involution in broiler hens.

Acetylcholinesterase expression mediated by c-Jun-NH2-terminal kinase pathway during anticancer drug-induced apoptosis.

It has been shown that acetylcholinesterase (AChE) expression was induced during apoptosis and the anti-sense oligonucleotides and siRNA of AChE may prevent apoptosis in various cell types. However, the mechanisms underlying AChE upregulation remain elusive. We demonstrated here that c-Jun NH2-terminal kinase (JNK) could mediate AChE expression. In this study, both etoposide and excisanin A, two anticancer agents, induced apoptosis in colon cancer cell line SW620 as determined by Annexin V staining, the cleavage of caspase-3 and the proteolytic degradation of poly (ADP-ribose) polymerase (PARP). The results showed that both the agents upregulated AChE in SW620 cells. In the meantime, JNK was also activated and the expression and phosphorylation of c-Jun increased in SW620 cells exposed to the two agents. The induced AChE mRNA and protein expression could be blocked by SP600125, a specific inhibitor of SAPK/JNK, and small interfering RNA directed against JNK1/2. Transfection with adenovirus-mediated dominant negative c-Jun also blocked the upregulation of AChE expression. Together, these results suggest that AChE expression may be mediated by the activation of JNK pathway during apoptosis through a c-Jun-dependent mechanism.

Senescence and telomere shortening induced by novel potent G-quadruplex interactive agents, quindoline derivatives, in human cancer cell lines.

Agents stabilizing G-quadruplexes have the potential to interfere with telomere replication by blocking the elongation step catalysed by telomerase or telomerase-independent mechanism and could therefore act as antitumor agents. In this study, we found that quindoline derivatives interacted preferentially with intramolecular G-quadruplex structures and were novel potent telomerase inhibitors. Treatment with quindoline derivatives reproducibly inhibited telomerase activity in human leukemia K562 cells and colon cancer SW620 cells. N'-(10H-Indolo [3,2-b] quinolin-11-yl)-N, N-dimethyl-propane-1,3-diamine (SYUIQ-5), (one of quindoline derivatives), when added to K562 and SW620 cell culture at nonacute cytotoxic concentrations, increased time of population doublings of K562 and SW620 cells, induced a marked cessation in cell growth and cellular senescence phenotype after 35 and 18 days, respectively. Growth cessation was accompanied by a shortening of telomere length, and induction of p16, p21 and p27 protein expression. However, another compound SYUIQ-7 with greater IC(50) for telomerase had no obvious cellular effect in nonacute cytotoxic concentrations. These results indicate that quindoline derivatives as novel potent G-quadruplex interactive agents induce senescence and telomere shortening in cancer cells and therefore are promising agents for cancer treatment.

Coencapsulation of hepatocytes and bone marrow stem cells: in vitro conversion of ammonia and in vivo lowering of bilirubin in hyperbilirubemia Gunn rats.

BACKGROUNDS/AIMS: This study investigates the ammonia removal capacity of coencapsulated hepatocytes and bone marrow stem cells in culture, and the treatment effect on hyperbilirubinemia Gunn rats when transplanted. METHODS: The hepatocytes and bone marrow stem cells isolated from Wistar rats were encapsulated alone or coencapsulated. In vitro, the encapsulated cells were cultured in media supplemented with 2.4 mMol/L concentration of ammonium chloride and the ammonia removal and urea synthesis were evaluated. In vivo, the encapsulated cells were transplanted intraperitoneally into hyperbilirubinemia Gunn rats and plasma bilirubin levels were measured before and after transplantation at intervals of 85 days. RESULTS: The ammonia removal capacity was maintained longer in the different ammonia concentration media in the coencapsulated hepatocytes and bone marrow cells culture. In the coencapsulation transplantation group, the plasma bilirubin levels were significantly lower than those in the group of hepatocytes encapsulation transplantation during the period of 3 to 10 weeks posttransplantion. CONCLUSIONS: The coencapsulated heaptocytes and bone marrow cells when compared to encapsulated hepatocytes could improve the maintenance of hepatocyte function both in vitro of ammonia removal in culture, and in vivo of the lowering the Gunn rats blood total bilirubin when transplanted.

Multiple drug resistance phenotype of human endothelial cells induced by vascular endothelial growth factor 165.

AIM: To investigate the effect of vascular endothelial growth factor 165 (VEGF165) on sensitivity of endothelial cells to anticancer drugs. METHODS: Human dermal microvessel endothelial cells (HDMEC) were incubated with anticancer drugs in the presence of VEGF165. Survival of endothelial cells was assayed by MTT method. DNA fragments of apoptosis were detected by agarose electrophoresis. Potential mechanisms underlying the effect of VEGF165 on endothelial cells were investigated with RT-PCR and Western blot analysis. RESULTS: VEGF165 induced the multidrug resistance phenotype of HDMEC to a wide variety of anticancer drugs such as epirubicin, cisplatin, etoposide, mytomycin C, vincristine, CPT-11, and taxol in vitro. This protective effect was partly due to the up-regulation of lung drug resistance protein (LRP) and multidrug resistance-associated protein (MRP), as well as the down-regulation of Bax protein induced by VEGF165. CONCLUSION: VEGF165 induced multidrug resistance phenotype of endothelial cells, which implicated the anti-angiogenic effect of anticancer drugs might depend on microenvironment of tumors in vivo.