A J-shaped relationship between the atherogenic index of plasma and new-onset myocardial infarction in hypertensive patients with obstructive sleep apnea: a cohort study.

OBJECTIVE: This study aimed to investigate the relationship between baseline atherogenic index of plasma (AIP) and new-onset myocardial infarction (MI) in hypertensive patients with obstructive sleep apnoea (OSA). PATIENTS AND METHODS: 2,281 participants were included in this analysis after strict adherence to the inclusion and exclusion criteria. Hazard ratio (HR) and 95% confidence interval (CI) were estimated using multivariable Cox regression models. A generalized additive model was employed to determine nonlinear relationships. RESULTS: In multivariate-adjusted models, there was a positive association between AIP and new-onset MI (per SD increase; HR=1.42, 95% CI: 1.22-1.65). Smoothing curve fitting revealed a J-shaped association between AIP and new-onset MI, with a turning point of approximately -0.08. The addition of AIP to a model with established risk factors improved the C-index (p=0.007), integrated discrimination improvement (p=0.007), and continuous net reclassification improvement (p=0.027) for the new-onset MI. CONCLUSIONS: A J-shaped relationship was observed between AIP and new-onset MI.

[Long-term efficacy evaluation of coated metal stent implantation for ureteroscopic lithotripsy related refractory ureteral stricture].

OBJECTIVE: To comprehensively evaluate the long-term efficacy and safety of coated metal stent implantation for ureteroscopic lithotripsy related refractory ureteral stricture (URL-rUS). METHODS: Electronic medical records of 30 patients (31 affected renal units) receiving coated metal stent implantation for URL-rUS from Sept. 2018 to Aug. 2021 at Peking University People' s Hospital were reviewed for analysis. Coated metal stents were implanted in retrograde approach via ureteroscope. Last outpatient follow-up was set as endpoint. Baseline information, stricture characteristics and decompression strategy before coated metal stent implantation were retrospectively collected. Serum creatinine (Scr) concentration and renal pelvis width (RPW) were used as renal function indicators. Peri-operative and long-term complications and according outcomes were recorded. For the patients who had double J tubes implanted for drainage before operation, Ureteral Stent Symptom Questionnaire (USSQ) was applied to evaluate the stent-related discomforts and quality of life (QoL) before and after surgery. Data analysis was conducted with SPSS (version 25.0; SPSS, Chicago, IL, USA). RESULTS: Totally 30 patients with 31 affected renal units were included. All the patients previously underwent unsuccessful traditional endoscopic balloon dilation or endoureterotomy before receiving coated metal stent implantation. The mean age was (45.5±9.3) years old. The median follow-up time were 14 (6-36) months. All coated metal stents were successfully placed with a median duration of 60 (30-195) min. No severe peri-operative complications occurred. At endpoint, 28 (90.3%) sides of coated metal stents remained unobstructed with a longest indwelling time of 36 months. Causes of failures included stent migration (1 case, 3.2%), encrustation (1 case, 3.2%) and repeated stent related urinary tract infection (1 case, 3.2%). When compared with the baseline, significant reductions were observed in both Scr concentration and RPW at endpoint [(90.7±19.5) mmol/L vs. (83.1±18.5) µmol/L, P < 0.01, for Scr; (2.5±1.3) cm vs. (1.9±0.8) cm, P < 0.01, for RPW], indicating good preservation of renal function and remission of hydronephrosis. For 26 patients with double J stents before operation, significant reduction of USSQ average score (100.4±6.6 vs. 82.1±4.9, P < 0.01) evidenced better life quality preserving ability of coated metal stent versus double J stent. CONCLUSION: Coated metal stent implantation is a safe and minimally invasive management of ureteral stricture providing a satisfying long-term patency rate, after which the patients' quality of life and renal function could be both improved. This method could serve as a promising alternative long-term maintenance treatment option for patients with URL-US, especially when traditional endoscopic interventions failed.

[Therapeutic effects of sacral neuromodulation on detrusor underactivity].

OBJECTIVE: To evaluate the effects of sacral neuromodulation (SNM) on detrusor underactivity (DUA). METHODS: From December 2019 to April 2020, 6 patients with DUA who had been treated with SNM were assessed retrospectively. The average age was 58 years (46-65 years), with 3 males and 3 females. All the patients were diagnosed with DUA by urodynamics examination. Obstruction of bladder outlet was excluded through the cystoscopy. No patient had the history of neurological disease. All the patients were placed with the bladder colostomy tube before SNM. One female patient accepted the trans-urethral resection of bladder neck. Two male patients accepted the trans-urethral resection of prostate. All the 3 patients had no improvement of void symptom after the urethral operation. Before SNM, the average 24 h times of voiding was 23.8 (18-33), average volume of every voiding was 34.2 mL (10-50 mL), average residual volume was 421.7 mL (350-520 mL). The preoperative and postoperative 24 h urine frequency, average voided volume, and average residual urine volume were compared respectively. RESULTS: Totally 6 patients underwent SNM with stage Ⅰ procedure. The operation time for stage Ⅰ procedure was 62-135 min (average 90 min). After an average follow-up of two weeks, stage Ⅱ procedure was performed on responders. Four patients accepted stage Ⅱ procedure (conversion rate 66.7%), the other two patients refused the stage Ⅱ procedure because the urine frequency did not reach the satisfied level. But all the patients had the improvement of residual urine volume. For the 4 patients at the follow-up of 10-15 months, the improvement of void was still obvious. For the all patients after stage Ⅰ procedure, the average 24 h urine frequency reduced to 13.5 times (9-18 times, P < 0.001), the average voided volume increased to 192.5 mL (150-255 mL, P < 0.001), and the average residual urine volume reduced to 97.5 mL (60-145 mL, P < 0.001). No adverse events, such as wound infection or electrode translocation were detected during an average follow-up of 11.3 months. Only one of the 4 patients who received the stage Ⅱ procedure did the intermittent catheterization for one time each day. CONCLUSION: SNM provides a minimal invasive approach for the management of DUA.

[Preparation and in vitro evaluation of fused deposition modeling 3D printed verapa-mil hydrochloride gastric floating formulations].

OBJECTIVE: To explore the feasibility of preparing gastric floating formulations by fused de-position modeling (FDM) 3D printing technology, to evaluate the in vitro properties of the prepared FDM 3D printed gastric floating formulations, and to compare the influence of different external shapes of the formulation with their in vitro properties. METHODS: Verapamil hydrochloride and polyvinyl alcohol (PVA) were used as the model drug and the excipient, respectively. The capsule-shaped and hemisphere-shaped gastric floating formulations were then prepared by FDM 3D printing. The infill percentages were 15%, the layer heights were 0.2 mm, and the roof or floor thicknesses were 0.8 mm for both the 3D printed formulations, while the number of shells was 3 and 4 for capsule-shaped and hemisphere-shaped formulation, respectively. Scanning electron microscopy (SEM) was used to observe the morpho-logy of the surface and cross section of the formulations. Gravimetric method was adopted to measure the weights of the formulations. Texture analyzer was employed to evaluate the hardness of the formulations. High performance liquid chromatography method was used to determine the drug contents of the formulations. The in vitro floating and drug release behavior of the formulations were also characterized. RESULTS: SEM showed that the appearance of the FDM 3D printed gastric floating formulations were both intact and free from defects with the filling structure which was consistent with the design. The weight variations of the two formulations were relatively low, indicating a high reproducibility of the 3D printing fabrication. Above 800.0 N of hardness was obtained in two mutually perpendicular directions for the two formulations. The drug contents of the two formulations approached to 100%, showing no drug loss during the 3D printing process. The two formulations floated in vitro without any lag time, and the in vitro floating time of the capsule-shaped and hemisphere-shaped formulation were (3.97±0.41) h and (4.48±0.21) h, respectively. The in vitro release of the two formulations was significantly slower than that of the commercially available immediate-release tablets. CONCLUSION: The capsule-shaped and hemisphere-shaped verapamil hydrochloride gastric floating formulations were prepared by FDM 3D printing technology successfully. Only the floating time was found to be influenced by the external shape of the 3D printed formulations in this study.

[Initial clinical experience and follow-up outcomes of treatment for ureteroileal anastomotic stricture with Allium coated metal ureteral stent].

OBJECTIVE: To summarize the initial clinical experience and follow-up results of the treatment for ureteroileal anastomotic stricture after radical cystectomy with Allium coated metal ureteral stent. METHODS: From September 2018 to September 2019, 8 patients with ureteroileal anastomotic stricture after radical cystectomy underwent Allium ureteral stent insertion in Peking University People's Hospital and People's Hospital of Daxing District. The preoperative renal pelvis width under ultrasound was collected to evaluate the postoperative hydronephrosis, creatinine and urea nitrogen (BUN) before and after surgery, perioperative infection, and stent-related complications. The serum creatinine and BUN, renal pelvis width under ultrasound, urography and abdominal plain film (KUB) were reviewed at the end of 1, 3, and 6 months and annually postoperatively to observe the stent position and morphology. The long-term stent patency rate, complication rate, renal function and hydronephrosis were followed up and analyzed. The t-test or rank-sum test was used to compare the measurement data of the matched sample from the preoperative to the last follow-up. RESULTS: In the study, 6 cases (7 sides) were ureteral ileal conduit stricture, and 2 cases (3 sides) ureteral orthotopic neobladder stricture. Before surgery, 5 patients underwent long-term indwelling of a single J ureteral stent, with an average indwelling time of (20.6±8.8) months and an average replacement frequency of (3.6±1.3) months/time. The mean width of renal pelvis was (26.5±9.1) mm on preoperative renal ultrasonography. Among them, 6 patients were successfully indwelled with Allium coated metal ureteral stent by retrograde approach, and 2 patients by combination of double-endoscopy and ante-retrograde approach. No surgery-related complications during perioperative period were observed. The mean follow-up period was 9.8 months and Allium stent and ureter remained unobstructed in all the patients at the last follow-up without replacement or removal. Compared with preoperative data, the mean width of renal pelvis and mean blood urea nitrogen (BUN) in the last follow-up period were significantly reduced [(26.5±9.1) mm vs. (13.4±2.5) mm, P=0.008; (11.6±2.3) mmol/L vs. (10.2±2.2) mmol/L, P=0.017], however, there were no significant differences in the average serum creatinine or hemoglobin (P>0.05). Ureteroileal anastomotic re-stricture and other stent-related complications were not observed in all the patients by antegrade urography. CONCLUSION: Allium coated metal ureteral stent could be used for the treatment for ureteroileal anastomotic stricture, which could maintain relatively long-term patency rate and protect renal function. The indwelling time was longer and it could improve quality of life of patients.

[Long-term analysis of safety and efficacy of standard percutaneous nephrolithotomy in patients with solitary kidneys].

OBJECTIVE: To evaluate the long-term efficacy and safety of ultrasound-guided percutaneous nephrolithotomy (PCNL) in the treatment of patients with solitary kidney stones. METHODS: The clinical data of 22 patients with solitary kidney stones treated with PCNL in Peking University People's Hospital from September 2008 to June 2014, with the follow-up data of more than 5 years were analyzed retrospectively. Perioperative indicators, postoperative stone free rate (SFR) and incidence of complications were recorded. Ultrasonography was used to evaluate the long-term stones recurrence rate. Serum creatinine and estimated glomerular filtration rate (eGFR) were used to assess the long-term renal function. RESULTS: In this group of 22 patients, the average age was (50.3±11.8) years, with 10 cases of anatomic solitary kidneys, 12 functional solitary kidneys, and the median stone diameter was 1.65 (1.1-3.9) cm. All the patients had multiple stones, including 7 cases of staghorn stones. The median pre-operative serum creatinine was 104.5 (60.0-460.0) µmol/L, and the mean eGFR was (60.3±29.4) mL/min, showing no statistically significant difference compared with that before surgery. The mean operative time was (88.2±42.0) min, and there were 11 cases of single-channel and double-channel PCNL. The median serum creatinine on the first day after surgery was 102.0 (63.0-364.0) µmol/L, and the mean eGFR was (58.0±25.1) mL/min. The mean postoperative hospital stay was (8.7±5.2) days. In this group, 5 patients (22.7%) presented short-term complications, among which 4 patients presented postoperative infection and massive hemorrhage at the same time, which improved after conservative treatment, and 1 patient presented pleural injury and improved after closed thoracic drainage. Two patients (9.1%) developed long-term complications, and ureteral stricture occurred 3 months after operation, which improved after balloon dilatation. The median follow-up time was 6.2 (4.7-11.1) years. The median serum creatinine at the last follow-up was 104.0 (72.4-377.0) µmol/L, and the mean eGFR was (60.1±23.7) mL/min, showing no statistically significant difference compared with that before surgery. Renal function decreased in 6 patients (27.3%). Initial and final SFR were 72.7% and 100%, respectively. During the 6.2-year follow-up, 9 patients (40.9%) experienced recurrence of kidney stone. After stone recurrence, 13 lithotomy surgeries were performed, and the SFR by the latest follow-up was 63.6%. CONCLUSION: This study had the longest follow-up time for patients with solitary kidney stones after PCNL reported at home and abroad. Ultrasound-guided standard PCNL was safe and effective in the treatment of solitary kidney stones. Long-term follow-up results showed that the recurrence rate of kidney stones was still high, but the long-term renal function was stable after operation, and some patients showed mild renal function decline.

[Progress in genomic DNA methylation of esophageal squamous cell carcinoma].

Abnormal genomic DNA methylation is an important epigenetic change in malignant tumors. Esophageal squamous cell cancer is one of common malignant tumors in our country. In this paper summarized and discussed the progress of genomic DNA methylation in the esophageal squamouscell cancer, including the level of genomic DNA methylation, frequent abnormally methylated genes, methylation markers and potential targets, etc. This paper might provide candidate biomarkers and targets for further studies on the mechanism of the tumorigenesis and development of the esophagealsquamouscell cancer, as well as for the clinical application of esophageal cancer.

Neuropilin-1 contributes to esophageal squamous cancer progression via promoting P65-dependent cell proliferation.

Neuropilin-1 (NRP1) is a non-kinase receptor recently implicated in tumor progression. Here we revealed that over-expression of NRP1 correlates with poor prognosis in esophageal squamous cell carcinoma (ESCC). NRP1-knockdown suppressed ESCC cell proliferation and xenograft tumor growth. Reduced NRP1 expression downregulated P65 mRNA and protein expression, and ectopic expression of P65-restored cell proliferation in NRP1-silenced cells. NRP1 regulates P65 transcription by activating cAMP responsive element binding protein (CREB). NRP1 interacted with and activated epidermal growth factor receptor (EGFR), and b1/b2 domain of NRP1 is responsible for the activation of EGFR. We also found that EGFR regulated CREB transcriptional activity via AKT. These data suggest that NRP1 is an upstream regulator in the P65-dependent proliferation signaling pathway and a candidate therapeutic target for ESCC.

[Clinical effect and safety of pegylated interferon-α-2b injection (Y shape, 40 kD) in treatment of HBeAg-positive chronic hepatitis B patients].

Objective: To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 µg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control. Methods: This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval (CI) were calculated, and non-inferiority was demonstrated if the lower limit of 95% CI was > -10%. The t-test, chi-square test, or rank sum test was used according to the types and features of data. Results: A total of 855 HBeAg-positive CHB patients were enrolled and 820 of them received treatment (538 in the trial group and 282 in the control group). The data of the full analysis set showed that HBeAg seroconversion rate at week 72 was 27.32% in the trial group and 22.70% in the control group with a rate difference of 4.63% (95% CI -1.54% to 10.80%, P = 0.1493). The data of the per-protocol set showed that HBeAg seroconversion rate at week 72 was 30.75% in the trial group and 27.14% in the control group with a rate difference of 3.61% (95% CI -3.87% to 11.09%, P = 0.3436). 95% CI met the non-inferiority criteria, and the trial group was non-inferior to the control group. The two groups had similar incidence rates of adverse events, serious adverse events, and common adverse events. Conclusion: In Peg-IFN-α regimen for HBeAg-positive CHB patients, the new drug Peg-IFN-α-2b (Y shape, 40 kD) has comparable effect and safety to the control drug Peg-IFN-α-2a.

[Clinical effect and safety of 144-week treatment with entecavir capsules in treatment-naïve HBeAg-positive patients with chronic hepatitis B].

Objective: To investigate the clinical effect and safety of entecavir capsules in the treatment of treatment-naïve HBeAg-positive patients with chronic hepatitis B (CHB). Methods: A total of 158 HBeAg-positive CHB patients were given oral entecavir capsules at a dose of 0.5 mg/time once a day for 144 weeks. Clinical outcome and safety were evaluated at baseline and at 24, 48, 72, 96, 120, and 144 weeks of treatment respectively. The Fisher's exact test was used for the analysis of categorical data. Results: After 144 weeks of treatment, 90.91% of all patients achieved virologic response (< 69 IU/ml), the normalization rate of alanine aminotransferase was 88.18%, the clearance rate of HBeAg was 33.33%, and the seroconversion rate of HBeAg was 24.07%. Of all patients, 2 dropped out due to adverse events and 5 experienced serious adverse reactions. Conclusion: Entecavir capsules can inhibit viral replication and have good safety in treatment-naïve HBeAg-positive CHB patients.

[Efficacy and safety of pegylated interferon α-2b injection (Y shape, 40 kD) in treatment of patients with genotype 1/6 chronic hepatitis C].

Objective: To investigate the efficacy and safety of the new investigational drug pegylated interferon α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 µg/week) combined with ribavirin in the treatment of patients with genotype 1/6 chronic hepatitis C (CHC), with standard-dose Peg-IFN-α-2a combined with ribavirin as a positive control. Methods: A multicenter, randomized, open-label, and positive-controlled phase III clinical trial was performed. Eligible patients with genotype 1/6 CHC were screened out and randomly divided into Peg-IFN-α-2b(Y shape, 40kD) group and Peg-IFN-α-2a group at a ratio of 2:1. The patients in both groups were given oral ribavirin for 48 weeks in addition and then followed up for 24 weeks after drug withdrawal. Abbott Real Time HCV Genotype II was used to determine HCV genotype, and Cobas TaqMan quantitative real-time PCR was used to measure HCV RNA level at 0, 4, 12, 24, 48, and 72 weeks. Adverse events were recorded in detail. The primary efficacy endpoint was sustained virological response (SVR), and a non-inferiority test was also performed. Results: A total of 561 patients with genotype 1/6 CHC were enrolled, among whom 529 received treatment; 90.9% of these patients had genotype 1 CHC. The data of the full analysis set showed that SVR rate was 69.80% (95% CI 65.00%-74.60%) in the trial group and 74.16% (95% CI 67.73%-80.59%) in the control group (P = 0.297 0). The data of the per protocol set (PPS) showed that SVR rate was 80.63% (95% CI 76.04%-85.23%) in the trial group and 81.33% (95% CI 75.10%-87.57%) in the control group (P = 0.849 8), and the 95% CI of rate difference conformed to the non-inferiority standard. The analysis of the PPS population showed that of all subjects, 47.9% achieved rapid virologic response, with a positive predictive value of 93.8%. The incidence rate of adverse events was 96.30% in the trial group and 94.94% in the control group, and the incidence rate of serious adverse events was 5.13% in the trail group and 5.06% in the control group. Conclusion: In the regimen of Peg-IFN-α combined with ribavirin for the treatment of genotype 1/6 CHC, the new investigational drug Peg-IFN-α-2b(Y shape, 40 kD) has comparable clinical effect and safety to the control drug Peg-IFN-α-2a.

[Genomic analysis of Wenzhou virus in rodents from Zhejiang province].

Objective: Arenavirus is a negative single-stranded RNA virus and an important human pathogen, mainly harbored and transmitted by rodents, causing severe diseases, including hemorrhagic fever and encephalitis. Following the discovery of a novel pathogenic arenavirus (Wenzhou virus, WENV), the prevalence of WENV in local small rodents was investigated. Methods: By using RT-PCR, WENV was screened in 48 and 156 rodents sampled from Wenzhou and Longquan, respectively. Results: Consequently, WENV was detected in 5 (10.41%) rodents sampled from Wenzhou. However, no WENV was identified in all the rodents sampled from Longquan. Genetic analysis of complete genome sequences indicated that 4 of 5 virus strains were closely related to the known Wenzhou viruses with high homology. Especially, the L and S segments of Wencheng-Rn-288 strain shared homology of 87.5% and 91.6% with other viruses, respectively. They formed a distinct lineage, suggesting that this strain might be a novel variant of WENV. Conclusions: Our results indicate that WENV has a high prevalence and high genetic diversity among rodents in Wenzhou. As the respiratory disease caused by WENV has been detected in Cambodia, it is necessary to strengthen the surveillance for WENV in China.

Integrative analyses of transcriptome sequencing identify novel functional lncRNAs in esophageal squamous cell carcinoma.

Long non-coding RNAs (lncRNAs) have a critical role in cancer initiation and progression, and thus may mediate oncogenic or tumor suppressing effects, as well as be a new class of cancer therapeutic targets. We performed high-throughput sequencing of RNA (RNA-seq) to investigate the expression level of lncRNAs and protein-coding genes in 30 esophageal samples, comprised of 15 esophageal squamous cell carcinoma (ESCC) samples and their 15 paired non-tumor tissues. We further developed an integrative bioinformatics method, denoted URW-LPE, to identify key functional lncRNAs that regulate expression of downstream protein-coding genes in ESCC. A number of known onco-lncRNA and many putative novel ones were effectively identified by URW-LPE. Importantly, we identified lncRNA625 as a novel regulator of ESCC cell proliferation, invasion and migration. ESCC patients with high lncRNA625 expression had significantly shorter survival time than those with low expression. LncRNA625 also showed specific prognostic value for patients with metastatic ESCC. Finally, we identified E1A-binding protein p300 (EP300) as a downstream executor of lncRNA625-induced transcriptional responses. These findings establish a catalog of novel cancer-associated functional lncRNAs, which will promote our understanding of lncRNA-mediated regulation in this malignancy.

[Expression and intratumoral heterogeneity of LN-5γ2 in esophageal squamous cell carcinoma].

OBJECTIVE: The present study aimed to evaluate the expression and intratumoral heterogeneity of LN-5γ2 in esophageal squamous cell carcinoma (ESCC). METHODS: The expression of LN-5γ2 protein was examined in 135 ESCC cases by immunohistochemistry, and to analyze its relationship with the clinical relevance of patients. The protein expressions in different regions in the same tumor as well as different nests in the same region were compared. RESULTS: Moderate and high expression of LN-5γ2 protein was detected in 40.0% (54/135) of tumor tissues. Positive immunohistochemical staining was observed in 31.1% (23/74) of early stage (stages Ⅰ/Ⅱ) cases and 50.8% (31/61) of late stage (stage Ⅲ) cases, with a significant difference between these two groups (P=0.023). There was no statistical association of LN-5γ2 expression with age, sex of patients, PT sage, lymph node metastasis and degree of tumor differentiation (P>0.05). However, differential expression of LN-5γ2 protein was found at different sampling sites in the same tumor and the same sampling site in different carcinomas. CONCLUSION: High expression of LN-5γ2 is positively correlated with tumor clinical stages and there existed intratumoral heterogeneity of LN-5γ2 expression in ESCC tissues.

[Association between ribavirin plasma concentration and sustained virologic response in treatment of patients with genotype 1b chronic hepatitis C with pegylated interferon-α-2b and ribavirin].

OBJECTIVE: To investigate the association between the dose and plasma concentration of ribavirin (RBV) and sustained virologic response (SVR) during the anti-hepatitis C virus (HCV) treatment with pegylated interferon-α-2b (PEG-IFN-α-2b) and RBV. METHODS: A total of 40 patients with chronic hepatitis C (CHC) who were treated with PEG-IFN-α-2b and RBV as the antiviral treatment were enrolled, and according to the therapeutic effect (SVR was defined as HCV RNA maintained below the lower limit of detection at 24 weeks after drug discontinuation in patients who achieved virologic response at the end of treatment, and recurrence was defined as HCV RNA turning positive), these patients were divided into SVR group (20 patients aged 19-55 years, including 10 male patients) and recurrence group (20 patients aged 21-76 years, including 12 male patients). The HPLC-MS/MS was used to measure the RBV plasma concentration at weeks 4, 12, 24, and 48 of treatment. The t-test and receiver operating characteristic (ROC) curve were used for statistical analysis. RESULTS: During the antiviral treatment, the dose of RBV showed a significant difference between the two groups (15.01 ± 1.21 mg/kg vs 10.28 ± 2.81 mg/kg,t= 6.908,P= 0.000). The area under the ROC curve reached 0.96 (95%CI0.00-1.00,P= 0.000), suggesting that the dose of RBV had a high value in predicting SVR. When the dose of RBV was higher than 13.05 mg/kg (sensitivity 100%; specificity 85%), the possibility of achieving SVR was also increased. The RBV plasma concentrations in the SVR group at weeks 4,12, 24, and 48 of treatment were 1 894.8 ± 740.7 ng/ml, 2 029.9 ± 547.7 ng/ml, 2 011.8 ± 354.2 ng/ml, and2 093.5 ± 540.3 ng/ml, respectively, and those in the recurrence group were 1 223.1 ± 722.7 ng/ml, 1 286.9±685.4 ng/ml, 1304.7 ± 692.0 ng/ml, and 1 221.3 ± 655.3 ng/ml, respectively. The RBV plasma concentration at each time point showed significant differences between the two groups (t= 2.903,P= 0.006;t= 3.787,P= 0.001;t= 4.068,P= 0.000;t= 4.593,P= 0.000). The results of ROC analysis showed that the areas under the ROC curve at weeks 4, 12, 24, and 48 of treatment were 0.76 (95%CI0.61-0.92,P= 0.005), 0.83 (95%CI0.68-0.97,P= 0.000), 0.83 (95%CI0.69-0.98,P= 0.000), and 0.86 (95%CI0.72-1.00,P= 0.000), respectively, suggesting that the RBV plasma concentration had a high value in predicting SVR. When the cut-off values of RBV plasma concentration at weeks 4, 12, 24, and 48 of treatment were higher than 1262.5 ng/ml (sensitivity 90%; specificity 60%), 1432 ng/ml (sensitivity 100%; specificity 65%), 1427 ng/ml (sensitivity 100%; specificity 65%), and 1610 ng/ml (sensitivity 95%; specificity 80%), respectively, there was a greater possibility of achieving SVR. CONCLUSION: During the antiviral treatment with PEG-IFN-α-2b and RBV, the dose and plasma concentration of RBV have a high value in predicting the recurrence of CHC and the possibility of SVR.

Microtubule-associated protein 4 is an important regulator of cell invasion/migration and a potential therapeutic target in esophageal squamous cell carcinoma.

Cell invasion and migration significantly contribute to tumor metastasis. Microtubule-associated protein 4 (MAP4) protein is one member of microtubule-associate proteins family. It is responsible for stabilization of microtubules by modulation of microtubule dynamics. However, there is little information about the involvement of MAP4 in human cancer. Here we show that MAP4 serves as a regulator of invasion and migration in esophageal squamous cancer cells. By activating the ERK-c-Jun-vascular endothelial growth factor A signaling pathway, MAP4 promotes cell invasion and migration in vitro, tumor growth and metastasis in mouse models. Immunohistochemical staining of operative tissues indicated that MAP4 expression was associated with tumor stage, lymph node metastasis and shorter survival of the patients with esophageal squamous cell carcinoma (ESCC). Multivariate Cox regression analysis showed that MAP4 is an independent prognostic indicator. In the serial sections of ESCC tissues, there was a positive correlation between MAP4 and vascular endothelial growth factor A expression. Notably, an intratumoral injection of MAP4-small interfering RNA (siRNA) remarkably inhibited the growth of the tumors that formed by the MAP4-expressing ESCC cells in nude mice, and a combination of MAP4-siRNA and Bevacizumab significantly enhanced the inhibition effect. Our data suggest that MAP4 is probably a useful prognostic biomarker and a potential therapeutic target for the disease.

Identification of putative target genes for amplification within 11q13.2 and 3q27.1 in esophageal squamous cell carcinoma.

BACKGROUND: Genomic aberration is a common feature of human cancers and also is one of the basic mechanisms that lead to overexpression of oncogenes and underexpression of tumor suppressor genes. Our study aims to identify frequent genomic changes and candidate copy number driving genes in esophageal squamous cell carcinoma (ESCC). METHODS: We used array comparative genomic hybridization to identify recurrent genomic alterations and screened the candidate targets of selected amplification regions by quantitative and semi-quantitative RT-PCR. RESULTS: Thirty-four gains and 16 losses occurred in more than 50 % of ESCCs. High-level amplifications at 7p11.2, 8p12, 8q24.21, 11q13.2-q13.3, 12p11.21, 12q12 and homozygous deletions at 2q22.1, 8p23.1-p21.2, 9p21.3 and 14q11.2 were also identified. 11q13.2 was a frequent amplification region, in which five genes including CHKA, GAL, KIAA1394, LRP5 and PTPRCAP were overexpressed in tumor tissues than paracancerous normal tissues. The expression of ALG3 at 3q27.1 was higher in ESCCs, especially in patients with lymph node metastasis. CONCLUSIONS: Target gene identification of amplifications or homozygous deletions will help to reveal the mechanism of tumor formation and explore new therapy method.

Calreticulin promotes cell motility and enhances resistance to anoikis through STAT3-CTTN-Akt pathway in esophageal squamous cell carcinoma.

We have shown earlier that overexpression of Calreticulin (CRT) contributed to a poor prognosis for patients with esophageal squamous cell carcinoma (ESCC). Here, we have shown an important role of CRT in tumorigenesis through enhancing cell motility and anoikis resistance. SiRNA-mediated knockdown of CRT caused impaired cell migration, invasion and resistance to anoikis. Notably, CRT downregulation decreased the expression of Cortactin (CTTN), which has been previously reported as a candidate oncogene associated with anoikis through the PI3K-Akt pathway. In addition, Akt phosphorylation was abolished after CRT downregulation and its activation can be refreshed by CRT upregulation, suggesting that CRT-enhanced cell resistance to anoikis through the CRT-CTTN-PI3K-Akt pathway. Moreover, the CTTN mRNA level was decreased in CRT-siRNA cells, coupled with the inactivation of STAT3. Expression of both CTTN and p-STAT3 was reduced in tumor cells following incubation with the JAK-specific inhibitor, AG490. Chromatin immunoprecipitation assay showed direct binding of p-STAT3 to the conservative STAT3-binding sequences in CTTN promoter. Furthermore, overexpression of CTTN in CRT-downregulated ESCC cells restored its motility and resistance to anoikis. This study not only reveals a role of CRT in motility promotion and anoikis resistance in ESCC cells, but also identifies CRT as an upstream regulator in the CRT-STAT3-CTTN-Akt pathway.

Protein alterations in ESCC and clinical implications: a review.

Esophageal squamous cell carcinoma (ESCC) is the predominant histological subtype of esophageal cancer in Asia, characterized by high incidence and mortality rate. Although significant progress has been made in surgery and adjuvant chemoradiotherapy, the prognosis of the patients with this cancer still remains poor. Investigation into protein alterations that occurred in tumors can provide clues to discover new biomarkers for improving diagnosis and guiding targeted therapy. Hundreds of papers have appeared over the past several decades concerning protein alterations in ESCC. This review summarizes all the dysregulated proteins investigated in the disease from 187 published papers and analyzes their contributions to tumor development and progression. We document protein alterations associated with tumor metastasis and the transition from normal esophageal epithelia to dysplasia in order to reveal the most useful markers for prediction of clinical outcome, early detection, and identification of high-risk patients for targeted therapies. In particular, we discuss the largest and most rigorous studies on prognostic implications of proteins in ESCC, in which cyclin D1, p53, E-cadherin and VEGF appeared to have the strongest evidence as independent predictors of patient outcome.

BRCA1 germline mutations in Chinese patients with hereditary breast and ovarian cancer.

The role of germline BRCA1 mutations in hereditary breast and ovarian cancer (HBOC) has been well established in women in Western countries. However, relatively few studies have been carried out in Chinese population. In the present study, we investigated the frequency and spectrum of germline BRCA1 mutations in Chinese HBOC patients, all of whom were from northern part of China. A total of 25 women with HBOC and ten relatives were analyzed. Mutation screening was performed by a combination of denaturing high-performance liquid chromatography and sequencing. Seven protein-truncating mutations were identified. They were 667delG, 3347A --> T, 3478del5, 4255delCT, 1235A --> G, 2064G --> T, and 5589del8. The first four of the mutations were putative ones never reported before. The prevalence of the protein-truncating mutations in this HBOC series was 40.0%, which is similar to that observed in Western hereditary ovarian cancer patients but higher than that reported in Chinese women with sporadic breast and ovarian cancer. Among the ten relatives we analyzed, six shared the same mutations with their affected relatives. No ovarian cancer was detected after 19 months of follow-up. This study showed that BRCA1 mutations play an important role in Northern Chinese HBOC.