[Treatment of stage 3b diabetic kidney disease patients with macroalbuminuria by qizhi jiangtang capsule: a multicenter randomized control clinical study].

OBJECTIVE: To observe the efficacy and safety of Qizhi Jiangtang Capsule (QJC) in treating stage 3b diabetic kidney disease (DKD) patients with macroalbuminuria. METHODS: Patients who conformed to the diagnostic criteria of stage 3b DKD were randomly assigned to two groups according to random digital table, the experiment group and the control group, 84 in each group. All patients received a two-week elution period, and then were treated with basic Western therapy. Patients in the experiment group took QJC, 5 pills per time, 3 times a day, while those in the control group took Valsartan Capsule 160 mg each time, once daily. The observation period of follow-ups was limited within 6 months, and the time points were set as the baseline, 1st month, 3rd month, and 6th month. Systolic blood pressure (SBP), diastolic blood pressure (DBS), 24 h urine protein quantitative (24 h UPQ), plasma albumin (ALB), and serum creatinine (SCr) were detected and recorded, and estimated glomerular filtration rate (eGFR) was calculated. The occurrence of hypoglycemic reaction, coagulation disorder, gastrointestinal tract reaction, allergy, hyperkalemia, doubling of creatinine, and overall adverse events were observed and recorded at same time. RESULTS: Finally 81 patients in the experiment group and 80 patients in the control group were effectively included. Compared with the baseline level, SBP and DBS obviously decreased in the control group at month 1 of treatment (P < 0.05), and more significantly decreased at month 6 of treatment (P < 0.01). SBP at month 1, 3, and 6 of follow-ups; DBS at month 6 of follow-ups was lower in the control group than in the experiment group (P < 0.05). At month 1, 3, and 6 of follow-ups, 24 h UPQ of the experiment group was significantly lower than the baseline level (P < 0.01). It was also significantly lower than the level of the control group at the same time point (P < 0.05). There was no significant difference in 24 h UPQ at month 1, 3, and 6 of follow-ups between the control group and the baseline level (P > 0.05). ALB of the experiment group showed an increasing trend. It was significantly higher than the baseline level at month 6 (P < 0.05), which was also higher than that of the control group at same period (P < 0.05). There was no significant difference in the ALB level in the control group (P > 0.05). SCr of two groups showed an increasing trend. SCr of the experiment group was significantly higher at month 1, 3, and 6 follow-ups than the baseline level (P < 0.05). But the increment of SCr was higher in the control group than in the experimental group, and obviously higher than the baseline levels (P < 0.05). eGFR of both groups showed a decreasing trend. The decrement was higher in the control group than in the experimental group (P < 0.05). The proportion of progression of renal functions at month 1, 3, and 6 of follow-ups in the experimental group was 0.0% (0 case), 9.55% (8 cases), and 21.4% (18 cases), while they were 8.3% (7 cases), 21.4% (18 cases), and 40.5% (34 cases) in the control group. There was no statistical difference in the proportion of progression of renal functions between the two groups at month 3 and 6 of follow-ups (P < 0.05). There was no statistical difference in the incidence of adverse reactions between two groups (P > 0.05). CONCLUSION: QJC could effectively reduce urinary protein of patients with stage 3b DKD, and delay the progression of renal functions.

Krüppel-like factor 10 expression as a prognostic indicator for pancreatic adenocarcinoma.

Deregulation of transforming growth factor (TGF)-ß function is a common feature of pancreatic cancer, rendering these cancers unresponsive to TGF-ß-stimulated growth inhibition. Recent findings have supported a primary role for Krüppel-like factor 10 (KLF10) as an important transcription factor involved in mediating TGF-ß1 signaling. The aim of this study was to evaluate the correlation between KLF10 expression and the clinical and pathologic features of pancreatic cancer. Tissue specimens from patients with pancreatic adenocarcinoma were retrospectively collected for immunohistochemical analysis. To demonstrate that Klf10 expression was primarily regulated by methylation status, the Klf10 promoter was examined by methylation-specific PCR using a pancreatic cancer cell line (Panc-1). DNA methyltransferase (DNMT) inhibitor and small-interfering RNA depletion of DNMT genes were used to reverse KLF10 expression in the Panc-1 cells. In parallel, DNMT1 expression was evaluated in the pancreatic cancer tissue specimens. In 95 pancreatic cancer tissue specimens, KLF10 expression was inversely correlated with pancreatic cancer stage (P = 0.01). Multivariable analysis revealed that, in addition to the presence of distant metastasis at diagnosis (P = 0.001 and 0.001, respectively), KLF10 was another independent prognostic factor related to progression-free and overall survival (P = 0.018 and 0.037, respectively). The loss of KLF10 expression in advanced pancreatic cancer is correlated with altered methylation status, which seems to be regulated by DNMT1. Our results suggest that KLF10 is a potential clinical predictor for progression of pancreatic cancer.

Diffuse sclerosing variant of thyroid papillary carcinoma: diagnostic challenges occur with Hashimoto's thyroiditis.

Diffuse sclerosing papillary thyroid carcinoma (DSPTC) is a relatively rare variant of papillary thyroid carcinoma with distinct histological features, radiological characteristics, and biological aggressiveness. Compared with conventional papillary thyroid carcinoma, DSPTC is characterized by scattered microscopic tumor islands, diffuse fibrosis, calcification, and abundant lymphocytic aggregation. A preoperative diagnosis is challenging in the absence of nodules and scanty fine needle aspiration cytology samples. We describe a unique DSPTC patient, an 18-year-old woman who presented with a neck mass that grew slowly for 2 years. The palpable neck mass was nontender, well defined, firm, and unmovable. Laboratory studies showed normal thyroid function and positive autoimmune markers: antithyroglobulin antibody = 1:1600 and antimicrosomal antibody = 1:1600. A neck ultrasound showed diffusely prominent microcalcifications with one small vague nodule. Hashimoto's thyroiditis with an accompanying malignancy was suspected. Based on the result of intraoperative pathology reports, the patient was given a total thyroidectomy. Lymph node dissection and histological analysis revealed bilateral DSPTC in addition to lymphocytic thyroiditis in nonmalignant areas of the thyroid. Clinical and histological diagnostic challenges usually occur when DSPTC presents with a diffuse thyroid enlargement, dispersed microscopic tumor islands (frequently without mass formation), extensive fibrosis, and abundant lymphocytic infiltration mimicking thyroiditis.

CDCP1 (CUB domain containing protein 1) is a potential urine-based biomarker in the diagnosis of low-grade urothelial carcinoma.

Urine-based cytology is non-invasive and widely used for clinical diagnosis of urothelial carcinoma (UC), but its sensitivity is less than 40% for low-grade UC detection. As such, there is a need for new diagnostic and prognostic biomarkers of UC. CUB domain containing protein 1 (CDCP1) is a type I transmembrane glycoprotein highly expressed in various cancers. Using tissue array analysis, we demonstrated that CDCP1 expression in UC patients (n = 133), especially in those with low-grade UC, was significantly higher than in 16 normal persons. In addition, CDCP1 expression in urinary UC cells could also be detected by using immunocytochemistry method (n = 11). Furthermore, in 5637-CD cells, overexpression of CDCP1 affected the expression of epithelial mesenchymal transition-related markers and increased matrix metalloproteinase 2 expression and migration ability. Conversely, the knockdown of CDCP1 in T24 cells had the opposite effects. Using specific inhibitors, we demonstrated the involvement of c-Src/PKCδ signaling in the CDCP1-regulated migration of UC. In conclusion, our data suggest that CDCP1 contributes to the malignant progression of UC and may have the potential as a urine-based biomarker for detecting low-grade UC. However, a cohort study needs to be conducted.

A case report of appendiceal adenocarcinoma without gastrointestinal evidence mimicking primary ovarian cancer.

Metastatic tumours to the ovary comprise 10-25% of ovarian malignancies and may originate from various primary sites. Here, the case of a 49-year-old female patient who presented with periumbilical nodules and abdominal bloating is reported. She was found to have bilateral ovarian tumours with peritoneal carcinomatosis and ascites. Primary ovarian cancer was suspected while no contributory gastrointestinal lesion was detected by imaging studies and endoscopic examinations. Three cycles of neoadjuvant chemotherapy were administered, followed by interval debulking surgery. Appendiceal cancer was highly suspected based on analysis of a frozen section obtained during surgical debulking. Following the pathology investigation, the patient was finally diagnosed with primary appendiceal adenocarcinoma. She underwent chemotherapy comprising irinotecan and fluorouracil. Due to disease progression despite several chemotherapy regimens, the patient declined further treatment and was lost to follow-up 1 year after the debulking surgery. Metastatic tumours to the ovary may mimic primary ovarian cancers and often present with nonspecific manifestations. Therefore, meticulous exploration of the primary site is warranted if the diagnosis is clinically suspicious.

Tubal metaplasia of the endometrium with cytologic atypia: analysis of p53, Ki-67, TERT, and long-term follow-up.

Tubal metaplasia of the endometrium may occasionally display cytologic atypia (atypical tubal metaplasia) resembling serous carcinoma or endometrial intraepithelial carcinoma. Although atypical tubal metaplasia is presumed to be reactive or degenerative in etiology, its clinical significance is unknown. In this study, we investigated atypical tubal metaplasia in regard to its immunoexpression of p53, Ki-67, and human telomerase reverse transcriptase (TERT), and its long-term clinical outcome. A total of 63 cases of atypical tubal metaplasia and 200 cases of endometrial samples with typical tubal metaplasia were followed for a mean of 64 and 61 months, respectively. Of the 63 atypical tubal metaplasia cases, formalin-fixed, paraffin-embedded tissue sections from 16 cases were immunostained with antibodies to p53, Ki-67, and TERT. Sections from 13 cases of uterine serous carcinoma were also stained for TERT as control. After long-term follow-up, 5% of patients in the atypical tubal metaplasia group developed hyperplasia without atypia compared with 4% of patients in the control group (P=0.44), whereas 3% in the atypical tubal metaplasia group developed atypical hyperplasia or carcinoma compared with 2% in the control group (P=0.44). p53 immunoreactivity was either focal and weak or negative in all cases of both atypical and typical tubal metaplasia (P>0.05). Ki-67 immunoreactivity was present in 0-5% of cells in 94% of both atypical and typical tubal metaplasia (P>0.05). TERT immunoexpression was absent in all 16 cases of atypical tubal metaplasia, but present in all 13 cases of uterine serous carcinoma (P<0.0001). Our study indicates that atypical tubal metaplasia displays an immunostaining pattern similar to otherwise typical tubal metaplasia of the endometrium, and distinct from uterine serous neoplasms. The presence of atypical tubal metaplasia in endometrial samplings does not increase the risk of developing endometrial hyperplasia or malignancy.

Anaplastic Thyroid Cancer Successfully Treated With Radiation and Immunotherapy: A Case Report.

OBJECTIVE: Anaplastic thyroid cancer (ATC) is a rare thyroid cancer subtype with a devastating prognosis. Novel treatment strategies are under investigation to improve the survival of patients with ATC. METHODS: We present a case of recurrent ATC treated with a combination of radiation therapy (RT) and pembrolizumab, a programmed death-1 inhibitor, with a durable complete response. RESULTS: A 63-year-old woman underwent total thyroidectomy and left neck lymph node dissection and was diagnosed with papillary carcinoma in December, 2017. She received radioiodine in April, 2018. However, a left neck mass was noted in April, 2018 with biopsy demonstrating ATC with 95% positivity for programmed death-ligand 1 immunostaining. Positron emission tomography showed fluorodeoxyglucose uptake in the left thyroid bed and multiple lymph nodes in the left retropharyngeal, left neck, and right upper paratracheal areas. Hypofractionated RT for the recurrent areas was initiated in August,2018, and concomitant pembrolizumab was given 2 days after RT. A total of 10 cycles of pembrolizumab (2 mg/kg) were given every 3 weeks. The computed tomography scan after completion of RT and 3 cycles of pembrolizumab showed shrinkage of the neck lymph nodes. The serial follow-up computed tomography scans showed further shrinkage of the lymph nodes, and there was no recurrence of ATC as of October, 2020. CONCLUSION: We describe an ATC case successfully treated with a combination of RT and pembrolizumab with a durable response of 26 months and acceptable toxicities. This result warrants further investigation of this combination regimen in the treatment of ATC.

Upregulating sirtuin 6 ameliorates glycolysis, EMT and distant metastasis of pancreatic adenocarcinoma with krüppel-like factor 10 deficiency.

Krüppel-like factor 10 (KLF10) is a tumor suppressor in multiple cancers. In a murine model of spontaneous pancreatic adenocarcinoma (PDAC), additional KLF10 depletion accelerated distant metastasis. However, Klf10 knockout mice, which suffer from metabolic disorders, do not develop malignancy. The mechanisms of KLF10 in PDAC progression deserve further exploration. KLF10-depleted and KLF10-overexpressing PDAC cells were established to measure epithelial-mesenchymal transition (EMT), glycolysis, and migration ability. A murine model was established to evaluate the benefit of genetic or pharmacological manipulation in KLF10-depleted PDAC cells (PDACshKLF10). Correlations of KLF10 deficiency with rapid metastasis, elevated EMT, and glycolysis were demonstrated in resected PDAC tissues, in vitro assays, and murine models. We identified sirtuin 6 (SIRT6) as an essential mediator of KLF10 that modulates EMT and glucose homeostasis. Overexpressing SIRT6 reversed the migratory and glycolytic phenotypes of PDACshKLF10 cells. Linoleic acid, a polyunsaturated essential fatty acid, upregulated SIRT6 and prolonged the survival of mice injected with PDACshKLF10. Modulating HIF1α and NFκB revealed that EMT and glycolysis in PDAC cells were coordinately regulated upstream by KLF10/SIRT6 signaling. Our study demonstrated a novel KLF10/SIRT6 pathway that modulated EMT and glycolysis coordinately via NFκB and HIF1α. Activation of KLF10/SIRT6 signaling ameliorated the distant progression of PDAC.Clinical Trial Registration: ClinicalTrials.gov. identifier: NCT01666184.

Lateral patch-clamping in a standard 1536-well microplate format.

Lateral patch-clamping has emerged as a chip-based platform for automation of the conventional patch-clamp technique, the 'gold' standard for studying cellular ion channels. The conventional technique, as it relies on skilled-maneuver of glass micropipettes to patch cells, is extremely delicate, low in throughput, and thus cannot be used for primary screening of compounds against ion channels. Direct integration of glass capillaries on silicon provides lateral junctions for automated trapping and patching of cells. We demonstrate here a method of scaling up the lateral junctions to a standard 1536-well microtiter plate format. A single unit of 1536-well plate has been formed here on a 9 mm by 9 mm microstructured silicon with the inclusive of 16 wells molded in a capping layer made of polydimethylsiloxane (PDMS). The silicon substrate provides integrated glass capillaries (total 12) and their associated microfluidic network. Each glass capillary has an independent access through a dedicated well in PDMS and leads to a centralized channel in which cell suspension can be delivered through one of the remaining 4 wells. The unit has been tested on RBL-1 cells by recording whole-cell activity from inwardly rectifying endogenous potassium channels. A revised test protocol has been prescribed to avoid inaccurate readings due to altered ionic composition of the recording buffer when a typical suction is applied to capture cells.

[Investigation of therapeutic mechanism of Weiweifang on experimental gastric ulcer in rats viewing from metabonomics].

OBJECTIVE: To investigate the therapeutic mechanism of Weiweifang (WWF, a Chinese herbal preparation) on gastric ulcer in rats viewing from metabonomics. METHODS: Wistar rats were made to gastric model by acetic acid cauterization and randomized into the model group, the spontaneously healing group and the three WWF treatment groups, and a group of normal rats was set for control. Metabolic spectra of gastric mucosa extraction of rats were acquired with gas chromatography-mass spectrometry (GC-MS) technique. After being pre-processing, data were subjected to partial least squares discriminant analysis (PLS-DA) to discover the biomarkers in rats of the normal group and the model group. The therapeutic effect of WWF on experimental gastric ulcer was assessed by principal component analyses (PCA), and its action of mechanism was explained viewing from the changes of biomarkers. RESULTS: Spectra of biomarkers, including organic acids, fatty acids, amino acids, etc. in model rats were statistically different to those in normal rats, which demonstrated that the energy and substance metabolisms were disordered in rats with gastric ulcer. WWF could cure gastric ulcer effectively by way of regulating the metabolism of gastric mucosa. CONCLUSION: The therapeutic mechanism of WWF on experimental gastric ulcer in rats is revealed integrally by metabonomics in this study, displaying prominently the characteristics of Chinese medicine multiple targets comprehensive therapy.

[Effect of total saponins of Panax notoginseng on urinary albumin in patients with chronic renal failure].

OBJECTIVE: To observe the curative effect of total saponins of Panax notoginseng (PNS) on chronic renal failure (CRF). METHODS: Sixty patients with CRF (non-uremic) were randomly divided into the experimental and the control groups, with 30 cases in each group. Patients in experimental group were given PNS extract Xueshuantong 0.45 g on the basis of the general symptomatic treatment, once a day. While the patients in the control group were treated with Bailing capsule of 1.0 g, three times a day. Total therapeutic courses were 2 months for both groups. The changes in renal function, hemoglobin, 24-hour urinary protein, parathyroid hormone (PTH), N-acutely-ß-D-glucosaminidase (NAG) were observed in two groups. RESULTS: After 2 months, the changes in serum creatinine (SCr), clearance rate of endogenous creatinine (CCr), blood urea nitrogen (BUN), uric acid (UA), hemoglobin, 24-hour urinary protein were improved in both groups, while the changes in CCr, BUN, hemoglobin, 24-hour urinary protein in the experimental group were more obvious [CCr (ml/s): 0.36±0.13 vs. 0.34±0.12, BUN (mmol/L): 15.66±9.05 vs. 20.32±8.30, hemoglobin (g/L): 101.2±9.4 vs. 95.4±8.7, 24-hour urinary protein (mg): 1 040±450 vs. 2 360±390, all P<0.05]. After treatment, NAG (U/L) were decreased significantly only in control group (18.2±9.8 vs. 28.9±12.0, P<0.05). CONCLUSION: PNS has a good therapeutic effect for the treatment of CRF (non-uremic). It possesses such therapeutic effects as improving the renal function, and lowering urine protein.

Amiodarone-induced thyrotoxicosis in a patient with autonomously functioning nodular goiter.

OBJECTIVE: To report on adverse reactions associated with amiodarone and propylthiouracil. CASE SUMMARY: A 64-year-old female with atrial fibrillation and nodular goiter progressed to overt thyrotoxicosis after receiving therapy with amiodarone 200 mg/day for less than 12 weeks. Thyroid scan revealed a hyperfunctioning nodule in the left lobe, while immunologic studies were negative for both thyroid peroxidase and thyroglobulin antibodies. The thyroid-stimulating hormone (TSH) receptor antibody level was transiently elevated. Propylthiouracil 100 mg 3 times/day was started after the withdrawal of amiodarone, but the patient developed severe generalized skin rash, fever, and leukocytosis after 4 weeks. Thyroidectomy was performed, and histopathology was compatible with type 1 amiodarone-induced thyrotoxicosis (AIT) associated with toxic nodular goiter. An objective causality assessment revealed that thyrotoxicosis was probably related to use of amiodarone. DISCUSSION: Amiodarone is an antiarrhythmic agent that may cause thyroid dysfunction. Differentiating between the 2 types of AIT is important for implementation of the correct therapeutic strategy. The transient elevation of TSH receptor antibodies in AIT complicated the diagnosis. As a rare subtype, type 1 AIT by nodular goiter may be associated with early AIT. Initiating thyroid function monitoring within 3 months of amiodarone therapy should be considered. CONCLUSIONS: Type 1 AIT caused by nodular goiter is rarely reported. Amiodarone should be avoided in such patients and subtotal thyroidectomy to remove the toxic nodule may be the treatment of choice.

[Effects of L-carnosine in preventing and treating rat cataract induced by sodium selenite].

OBJECTIVE: To explore the effect of L-carnosine in preventing and treating rat cataract induced by sodium selenite. METHODS: This was an experimental study. Cataract was induced in the rats by sodium selenite. Rats were divided into 3 groups and each group has 3 ones. L-carnosine eye drops (50 g/L or 20 g/L ) and 0.9% normal saline were respectively instilled to the rat eye for 3 weeks and examined. L-carnosine was added to the medium of cultured rat cataract lens at 1.00, 0.10 and 0.01 g/L for 1 week and then examined. Cataract lens were studied by using two-dimensional electrophoresis. RESULTS: One week after instillation of L-carnosine, the scores of rat cataract were 2.22 +/- 0.65, 2.39 +/- 0.98 and 2.83 +/- 0.38 in 50 g/L, 20 g/L L-carnosine and control groups, respectively. The lesion in 50 g/L L-carnosine group was lighter than that of the control group. There was significant difference between these two groups (P = 0.013). On the 2nd and 3rd weeks, there was no difference among these three groups. After 1 week for culturing cataract lens, there was no difference between these three groups. The results of two-dimensional electrophoresis showed that the protein number was 182 and 161 in the L-carnosine and control groups, respectively. High molecular weight protein decreased and low molecular weight protein increased in L-carnosine group. CONCLUSIONS: L-carnosine at 50 g/L could restrain the development of early stage rat cataract. L-carnosine could modulate the rat lens protein in vitro, but could not affect lens cataract scores.

Vitamin D-Binding Protein Enhances Epithelial Ovarian Cancer Progression by Regulating the Insulin-like Growth Factor-1/Akt Pathway and Vitamin D Receptor Transcription.

Purpose: Malignant ascites of epithelial ovarian cancer (EOC) helps identify prognostic biomarkers or mechanisms of tumor progression. Vitamin D-binding protein (DBP) was revealed to be upregulated in EOC ascites in our previous proteomic study. Here, we examined the role of DBP in EOC.Experimental Design: We analyzed ascites, serum, and tissue samples of patients with newly diagnosed EOC to determine the prognostic effects of DBP. We verified DBP function using orthotopic animal models and DBP regulation in ovarian cancer cell lines.Results: Elevated ascitic DBP was significantly associated with poor response to chemotherapy, short progression-free interval, increased cancer progression, and death. Ascitic DBP overexpression was an independent unfavorable biomarker for progression-free survival; DBP overexpression in cancerous tissue was significantly related to chemoresistance. In vivo and in vitro investigations demonstrated an important role for DBP in ovarian cancer progression. Orthotopic model mice inoculated with DBP knockdown ovarian cancer cells displayed a significant reduction in tumor formation, malignant cell number, ascitic DBP levels, invasiveness, and metastasis, and increased survival compared with controls. In presence of vitamin D receptor (VDR), DBP promoted cell aggression (invasion and doubling time) via activation of the insulin-like growth factor-1/insulin-like growth factor-binding protein-2/Akt axis, and induced suppression of vitamin D-responsive genes. A NF-κB p65-binding site in the VDR promoter was identified as a major determinant of DBP-dependent VDR promoter activation.Conclusions: This study highlights the importance of DBP in ovarian tumor progression and the potential application of DBP as a therapeutic target for EOC. Clin Cancer Res; 24(13); 3217-28. ©2018 AACR.

[Identification of Evodia rutaecarpa (Juss.) benth and quantitative determination of the total alkaloid of rhizoma coptidis in yuhuanglian extract].

OBJECTIVE: To identify Eevodia rutaecarpa (Juss.) Benth and determine the contents of the total alkaloid of Rhizoma Coptidis in Yuhuanglian extract. METHODS: Thin layer chromatography (TLC) was used for the identification of Evodia rutaecarpa (Juss.) Benth with a mixed solvent of cyclohexane-ethyl acetate-methanol-triethylamine (19:5:1:1) as the developer and spraying with 10% sulfuric acid ethanol solution as colorimetric method. The contents of the total alkaloid of Rhizoma Coptidis was determined by UV spectrophometry in Yuhuanglian extract and the detection wavelength was 349 nm. RESULTS: Special spots could be observed at 365 nm on positions corresponding to Evodia rutaecarpa (Juss.) Benth, and the preparation, while the negative control showed no spots. The linear range of berberine hydrochloric was 1.04-11.44 microg/ml, r = 0.9999; the average recovery was 99.2%, and RSD was 1.0% (n=5). CONCLUSIONS: Evodia rutaecarpa (Juss.) Benth can be identified simply and fast using TLC and the method to determine the contents of the total alkalid of Rhizoma Coptidis by UV spectrophometry is accurate, rapid and simple. It can be used for quality control of Yuhuanglian extract.

Krüpple-like factor 10 regulates radio-sensitivity of pancreatic cancer via UV radiation resistance-associated gene.

BACKGROUND AND PURPOSE: Krüpple-like factor 10 (Klf10), an early response gene of TGFß, was reported to be a prognostic biomarker for pancreatic cancer survival. The role of Klf10 in predicting tumor response to cancer treatment is unknown. MATERIALS AND METHODS: Genetically manipulated MiaPaCa and Panc-1 cells were established to evaluate clonogenic survival, autophagy, apoptosis and DNA repair after radiation. The interaction between Klf10 and UV radiation resistance-associated gene (UVRAG) was demonstrated by ChiP-PCR and luciferase reporter assay. Orthotopic murine tumor model and clinical specimens were used to evaluate radio-sensitivity of pancreatic cancer. RESULTS: We found Klf10 silencing correlates with enhanced pancreatic cancer clonogenic survival and murine tumor growth after radiation. UVRAG was an essential down-stream mediator transcriptionally suppressed by Klf10. Silencing UVRAG mRNA in Klf10 depleted Panc-1 cells reversed the radio-resistant phenotypes including decreased apoptosis and enhanced DNA repair as well as autophagy. Metformin, an anti-diabetic agent, was found to increase Klf10 and suppress UVRAG expression to improve radiation cytotoxicity in pancreatic cancer. The predictive value of Klf10 in radiation response and the inverse correlation with UVRAG were confirmed in cohorts of pancreatic cancer patients. CONCLUSIONS: Klf10 is a potential biomarker in predicting and sensitizing radiation effect in pancreatic cancer.

Effect of Parecoxib as an Adjunct to Patient-Controlled Epidural Analgesia after Abdominal Hysterectomy: A Multicenter, Randomized, Placebo-Controlled Trial.

OBJECTIVE: This multicenter, randomized, placebo-controlled study evaluated the efficacy and side effects of parecoxib during patient-controlled epidural analgesia (PCEA) after abdominal hysterectomy. METHODS: A total of 240 patients who were scheduled for elective abdominal hysterectomy under combined spinal-epidural anesthesia received PCEA plus postoperative intravenous parecoxib 40 mg or saline every 12 h for 48 h after an initial preoperative dose of parecoxib 40 mg or saline. An epidural loading dose of a mixture of 6 mL of 0.25% ropivacaine and 2 mg morphine was administered 30 min before the end of surgery, and PCEA was initiated using 1.25 mg/mL ropivacaine and 0.05 mg/mL morphine with a 2-mL/h background infusion and 2-mL bolus with a 15-min lockout. The primary end point of this study was the quantification of the PCEA-sparing effect of parecoxib. RESULTS: Demographic data were similar between the two groups. Patients in the parecoxib group received significantly fewer self-administrated boluses (0 (0, 3) vs. 7 (2, 15), P < 0.001) and less epidural morphine (5.01 ± 0.44 vs. 5.95 ± 1.29 mg, P < 0.001) but experienced greater pain relief compared with the control group (P < 0.001). Patient global satisfaction was higher in the parecoxib group than the control group (P < 0.001). Length of hospitalization (9.50 ± 2.1, 95% CI 9.12~9.88 vs. 10.41 ± 2.6, 95% CI 9.95~10.87, P = 0.003) and postoperative vomiting (17% vs. 29%, P < 0.05) were also reduced in the parecoxib group. There were no serious adverse effects in either group. CONCLUSION: Our data suggest that adjunctive parecoxib during PCEA following abdominal hysterectomy is safe and efficacious in reducing pain, requirements of epidural analgesics, and side effects. TRIAL REGISTRATION: ClinicalTrials.gov (NCT01566669).

Epidermal Growth Factor Receptor Mutation and Anaplastic Lymphoma Kinase Gene Fusion: Detection in Malignant Pleural Effusion by RNA or PNA Analysis.

Analyzing EGFR mutations and detecting ALK gene fusion are indispensable when planning to treat pulmonary adenocarcinoma. Malignant pleural effusion (MPE) is a devastating complication of lung cancer and sometimes the only source for mutation analysis. The percentage of tumor cells in the pleural effusion may be low; therefore, mutant enrichment is required for a successful analysis. The EGFR mutation status in MPE was determined using three methods: (1) PCR sequencing of genomic DNA (direct sequencing), (2) mutant-enriched PCR sequencing of genomic DNA using peptide nucleic acid (PNA-sequencing), and (3) PCR sequencing of cDNA after reverse transcription for cellular RNA (RNA-sequencing). RT-PCR was also used to test cases for ALK gene fusion. PNA-sequencing and RNA-sequencing had similar analytical sensitivities (< 1%), which indicates similar enrichment capabilities. The clinical sensitivity in 133 cases when detecting the common EGFR exon 19 and exon 21 mutations was 56.4% (75/133) for direct sequencing, 63.2% (84/133) for PNA-sequencing, and 65.4% (87/133) for RNA-sequencing. RT-PCR and sequencing showed 5 cases (3.8%) with ALK gene fusion. All had wild-type EGFR. For EGFR analysis of MPE, RNA-sequencing is at least as sensitive as PNA-sequencing but not limited to specific mutations. Detecting ALK fusion can be incorporated in the same RNA workflow. Therefore, RNA is a better source for comprehensive molecular diagnoses in MPE.

[Inhibiting proliferation of cultured rat lens epithelial cells by bFGF antisense oligonucleotides and their liposomes].

OBJECTIVE: To evaluate the effects of basic fibroblast growth factor (bFGF) antisense oligonucleotides and their liposomes on the proliferation of cultured rat lens epithelial cells (LECs). METHODS: The rat LECs were cultured and bFGF antisense oligonucleotides (AONs), sense oligonucleotides (SONs), AON liposomes, SON liposomes were added to second passage of cells supplied with DMEM while empty liposomes served as controls. MTT assay was used to examine the proliferation of LECs, and RT-PCR was performed to quantify bFGF mRNA expression in LECs 24 h after treatment. RESULTS: The Shapes of rat LECs were polygonal after 24 h in culture. Cell confluence was reached in 2 - 3 weeks. After subculture, confluence was occurred in 1 - 2 weeks. The results of MTT assay were showed that the absorption (A) value of bFGF AONs was 0.138 +/- 0.074, and that of bFGF SONs and DMEM control were 0.325 +/- 0.097 and 0.370 +/- 0.079, respectively. The A value of AONs was significantly less than SONs and DMEM alone (P = 0.024, P = 0.005). The absorption (A) value of bFGF AON liposomes was 0.128 +/- 0.032, and that of bFGF SON liposomes and liposomes alone were 0.238 +/- 0.120 and 0.348 +/- 0.017. The absorption (A) value of AON liposomes was not significantly different from that of liposomes negative control (P = 0.000). By RT-PCR, the amounts of PCR product for bFGF AONs, SONs and DMEM control were 0.33 micro g, 0.99 micro g, 0.85 micro g. The amounts of PCR product of bFGF AON liposomes, SON liposomes and liposomes only were 0.23 micro g, 0.48 micro g, 0.56 micro g. CONCLUSIONS: The proliferation of cultured rat LECs is inhibited by the treatment of the antisense oligonucleotides, the inhibition is correlated with decreased expression of bFGF mRNA.