CPL-01, an investigational long-acting ropivacaine, demonstrates safety and efficacy in open inguinal hernia repair.

BACKGROUND: There is an unmet medical need for effective nonopioid analgesics that can decrease pain while reducing systemic opioid use. CPL-01, an extended-release injectable formulation of ropivacaine, is designed to safely provide analgesia and reduce or eliminate opioid use in the postoperative period. METHODS: Subjects undergoing open inguinal hernia with mesh were prospectively randomized to 1 of 3 doses of CPL-01 (10, 20, or 30 ml of 2% CPL-01, n = 14, 12, and 14, respectively), Naropin (150 mg, n = 40), or saline placebo (n = 13) infiltrated into the surgical site prior to closure. Pain and rescue medication usage was assessed, and Numeric Rating Scale (NRS) pain scores were adjusted for opioid usage using windowed worst observation carried forward (wWOCF) imputation. The primary efficacy endpoint was the mean area under the curve (AUC) of the NRS pain intensity scores with activity. RESULTS: Ninety-three subjects were treated, and 91 subjects completed 72 h of post-operative monitoring. Subjects who received the highest dose of CPL-01 in Cohort 3 showed a clinically meaningful reduction in postoperative pain intensity scores, which was the lowest value for any treatment in all cohorts, showing a trend towards statistical significance as compared to the pooled placebo group (p = 0.08), and numerically better than the 40 subjects who received Naropin. Opioid use through 72 h in subjects who received CPL-01 in Cohort 3 was approximately half of that shown in the placebo and Naropin groups; approximately 2/3 of the CPL-01 subjects (9/14) required no opioids at all through the first 72 h after the operation. More CPL-01 subjects avoided severe pain and were ready for discharge earlier than other groups. CPL-01 was safe and well-tolerated, with no clinically meaningful safety signals, and showed predictable and consistent extended-release pharmacokinetics. CONCLUSION: Results suggest that CPL-01 may be the first long-acting ropivacaine to address postoperative pain while reducing the need for opioids.

[Analysis of the predictive value of plateletcrit for the degree of liver fibrosis in patients with chronic HBV infection with ALT less than two times the upper limit of normal].

Objective: Plateletcrit (PCT) is considered a new potential index to predict the degree of liver fibrosis in patients with chronic hepatitis B (CHB). This study aimed to explore the predictive value of PCT for the degree of liver fibrosis in patients with chronic hepatitis B virus (HBV) infection with alanine aminotransferase (ALT) < 2× upper limit of normal (ULN). Measurement data were compared using the t-test, ANOVA, or non-parametric test (Mann-Whitney U test). Categorical variables were compared using χ (2) test or Fisher's exact test. Methods: 140 cases with chronic HBV infection who underwent liver biopsy and ALT < 2×ULN were enrolled from January 2016 to March 2021. Univariate and multivariate logistic regression and the area under the receiver operating characteristic curve (AUC) were used to determine the predictive value of PCT for the degree of liver fibrosis. The likelihood ratio (LR) was used to optimize the selection of the diagnostic cut-off. Results: (1) Among the 140 cases, there were 34 (24.3%) cases in the S0 stage, 47 (33.6%) cases in the S1 stage, 16 (11.4%) cases in the S2 stage, 19 (13.6%) cases in the S3 stage, and 24 (17.1%) cases in the S4 stage. The overall mean PCT level was 0.19 ± 0.06%. (2) Univariate analysis revealed that PCT between patients with stages of liver fibrosis was S(0-1) and S(2-4) (0.20% ± 0.05% vs. 0.16% ± 0.06%, t = 3.955, P < 0.001), S(0 -2) and S(3-4) (0.20% ± 0.05% vs. 0.15% ± 0.06%, t = 5.631, P < 0.001) and S(0-3) and S4 (0.20% ± 0.05% vs. 0.12% ± 0.05%, t = 7.113, P < 0.001), respectively, and the differences were statistically significant. Multivariate logistic regression analysis showed that PCT was an independent risk factor for liver fibrosis stages S(2-4), S(3-4), and S4 (OR = 0.925, 95% CI: 0.859 - 0.997, P = 0.042; OR = 0.867, 95% CI: 0.789 - 0.954, P = 0.003; OR = 0.708, 95% CI: 0.593 - 0.846; P < 0.001). (3) The AUCs of PCT were 0.702, 0.777, and 0.885 for diagnosing liver fibrosis stages S(2-4), S(3-4), and S4 in patients with chronic HBV infection with ALT < 2×ULN. PCT was superior for the cirrhosis (S4) diagnosis. 92 (65.7%) cases were diagnosed as cirrhosis or non-cirrhosis according to the LR optimized diagnostic and exclusion diagnostic cut-offs (≤0.09%, ≤0.17%), with an accuracy of 97.8%. Conclusion: PCT has a high diagnostic and exclusion value for cirrhotic patients with chronic HBV infection with ALT < 2×ULN. Furthermore, it can be used as a non-invasive diagnostic index for determining and assisting the diagnosis of cirrhosis in resource-constrained areas, reducing the need for pathological examination of liver biopsies, and it has the advantage of being simple and intuitive without complex calculations.

[Functional outcomes of 100 patients with adenocarcinoma of the esophagogastric junction undergoing Cheng's GIRAFFE(®) reconstruction after proximal gastrectomy].

Objective: To investigate the functional outcomes and postoperative complications of Cheng's GIRAFFE reconstruction after proximal gastrectomy. Methods: A descriptive case series study was conducted. Clinical data of 100 patients with adenocarcinoma of the esophagogastric junction who underwent Cheng's GIRAFFE reconstruction after proximal gastrectomy in Cancer Hospital of University of Chinese Academy of Sciences (64 cases), Zhejiang Provincial Hospital of Chinese Medicine (24 cases), Lishui Central Hospital (10 cases), Huzhou Central Hospital (1 case) and Ningbo Lihuili Hospital (1 case) from September 2017 to June 2021 were retrospectively analyzed. Of 100 patients, 64 were males and 36 were females; the mean age was (61.3 ± 11.1) years and the BMI was (22.7±11.1) kg/m(2). For TNM stage, 68 patients were stage IA, 24 were stage IIA and 8 were stage IIB. Postoperative functional results and postoperative complications of radical gastrectomy with Giraffe reconstruction were analyzed and summarized. Gastroesophageal reflux disease questionnaire (RDQ) score and postoperative endoscopy were used to evaluate the occurrence of reflux esophagitis and its grade (grade N, grade A, grade B, grade C, and grade D from mild to severe reflux). The continuous data conforming to normal distribution were expressed as (mean ± standard deviation), and those with skewed distribution were presented as median (Q1, Q3). Results: All the 100 patients successfully completed R0 resection, including 77 patients undergoing laparoscopic surgery and 23 patients undergoing laparotomy. The Giraffe anastomosis time was (38.6±14.0) min; the blood loss was (73.0±18.4) ml; the postoperative hospital stay was 9.5 (8.2, 13.0) d; the hospitalization cost was (6.0±0.3) ten thousand yuan. Fourteen cases developed perioperative complications (14.0%), including 7 cases of pleural effusion or pneumonia, 3 cases of anastomotic leakage, 2 cases of gastric emptying disorder, 1 case of gastrointestinal hemorrhage and 1 case of anastomotic stenosis, who were all improved and discharged after symptomatic management. Patients were followed up for (33.3±1.6) months. Eight patients were found to have reflux symptoms by RDQ scale six months after surgery, and 11 patients (11/100,11.0%) were found to have reflux esophagitis by gastroscopy, including 6 in grade A, 3 in grade B, and 2 in grade C. All the patients could control their reflux symptoms with behavioral guidance or oral PPIs. Conclusion: Cheng's GIRAFFE reconstruction has good anti-reflux efficacy and gastric emptying function; it can be one of the choices of reconstruction methods after proximal gastrectomy.

Risk factors for disease progression in patients with mild to moderate coronavirus disease 2019-a multi-centre observational study.

OBJECTIVES: Since December 2019, the novel coronavirus disease 2019 (COVID-19) that emerged in Wuhan city has spread rapidly around the world. The risk for poor outcome dramatically increases once a patient progresses to the severe or critical stage. The present study aims to investigate the risk factors for disease progression in individuals with mild to moderate COVID-19. METHODS: We conducted a cohort study that included 1007 individuals with mild to moderate COVID-19 from three hospitals in Wuhan. Clinical characteristics and baseline laboratory findings were collected. Patients were followed up for 28 days for observation of disease progression. The end point was the progression to a more severe disease stage. RESULTS: During a follow up of 28 days, 720 patients (71.50%) had recovered or were symptomatically stable, 222 patients (22.05%) had progressed to severe disease, 22 patients (2.18%) had progressed to the critically ill stage and 43 patients (4.27%) had died. Multivariate Cox proportional hazards models identified that increased age (hazard ratio (HR) 2.56, 95% CI 1.97-3.33), male sex (HR 1.79, 95% CI 1.41-2.28), presence of hypertension (HR 1.44, 95% CI 1.11-1.88), diabetes (HR 1.82, 95% CI 1.35-2.44), chronic obstructive pulmonary disease (HR 2.01, 95% CI 1.38-2.93) and coronary artery disease (HR 1.83, 95% CI 1.26-2.66) were risk factors for disease progression. History of smoking was protective against disease progression (HR 0.56, 95% CI 0.34-0.91). Elevated procalcitonin (HR 1.72, 95% CI 1.02-2.90), urea nitrogen (HR 1.72, 95% CI 1.21-2.43), α-hydroxybutyrate dehydrogenase (HR 3.02, 95% CI 1.26-7.21) and D-dimer (HR 2.01, 95% CI 1.12-3.58) at baseline were also associated with risk for disease progression. CONCLUSIONS: This study identified a panel of risk factors for disease progression in individuals with mild to moderate COVID-19.

MicroRNA-22 targets FMNL2 to inhibit melanoma progression via the regulation of the Wnt/ß-catenin signaling pathway and epithelial-mesenchymal transition.

OBJECTIVE: Melanoma is regarded as one common malignancy in skin cancers, and there is growing evidence that microRNAs (miRNAs) play a vital role in the oncogenesis of tumors. This study aimed to investigate the roles and mechanism of miR-22 in melanoma. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was utilized to detect the expressions of miR-22 and mRNA. The functions of miR-22 in melanoma cell proliferation, migration and invasion were investigated with functional assays, including MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) and transwell assay. Western blots were utilized to examine the protein expressions. Luciferase reporter analysis was conducted to confirm the interactions between formin-like 2 (FMNL2) and miR-22 in melanoma cells. FMNL2 expression levels in melanoma tissues were investigated by immunohistochemistry (IHC) assays. RESULTS: The qRT-PCR analysis demonstrated significant decreased miR-22 expressions in melanoma tissues. Decreased miR-22 in melanoma tissues were correlated with adverse clinicopathologic features and poor prognosis. Functional assays indicated that upregulation inhibited melanoma cell proliferation, invasion and migration capacities. Luciferase reporter assays showed that FMNL2 was targeted by miR-22 in melanoma cells. Western blots indicated that miR-22 exerted anti-tumor functions by regulating the Wnt/ß-catenin and epithelial-mesenchymal transition (EMT). CONCLUSIONS: Our findings showed that miR-22 served as a tumor suppressor in melanoma progression, implying that miR-22 may function as a novel therapeutic target and prognostic biomarker for melanoma treatments.

[Analysis of 15 cases of avian influenza virus (H7N9) infection].

Objective: To describe the clinical, chest imaging, pathological manifestations and therapeutic experience of human infection with A/H7N9 virus. Methods: The features of 15 laboratory-confirmed cases of human infection with A/H7N9 virus in Taizhou, Jiangsu Province were retrospectively analyzed. Results: The 15 patients with confirmed viral pneumonia included 12 males and 3 females, with a median age of 61 years(ranging from 33 to 81 years). Twelve patients had a history of exposure to the poultry trading places, or direct contact with ill/dead avian, while 3 patients denied exposure or contact. The most common initial symptoms were fever, coughing, and respiratory distress. The illness progressed rapidly to acute respiratory distress syndrome (ARDS). Lab tests showed normal (8 cases) or decreased (7 cases)white blood cell count , decreased (13 cases) lymphocyte count and proportion , increased creatine kinase (CK, 12 cases) and lactate dehydrogenase (LDH, 15 cases), and respiratory failure (13 cases). Chest radiographic examination showed that the most common features were inflammatory infiltration in the lung, with partial consolidation. The average time of the diagnosis with influenza viral nucleic acid and onset of an oral anti-influenza drug were 7.1 days and 6.5 days. All patients were treated by antiviral drugs (oral oseltamivir 150 mg q12 h and/or intravenous paramivir 600 mg qd), with mechanical ventilation in 9 cases, glucocorticoid therapy in 5 cases (intravenous methylprednisolone in 3 and dexamethasone in 2 patients), extracorporeal membrane oxygenation (ECMO) therapy in 2 cases, continuous renal replacement therapy (CRRT) in 6 cases, and artificial liver therapy in 1 case. The pulmonary pathology was observed from post-mortem biopsy for 2 fatal cases. Patient 1 had diffuse alveolar damage with inflammatory exudation, hyaline membrane formation, and cellular infiltration. Patient 2 had widened alveolar septum, lymphocyte and monocyte cell infiltration in the alveolar septa, and interstitial fibrous proliferation. Nine patients were discharged, and 6 died. Conclusions: Patients with influenza A/H7N9 virus mostly presented with fever, cough, and were prone to progression to viral pneumonia. Once acute respiratory distress and important organ dysfunction occurred, the fatality rate was higher. Early diagnosis and rational treatment were critical for better outcomes.

[Clinical study of a new wearable device for rehabilitation after total knee arthroplasty].

Objective: To evaluate the effect of promoting knee joint rehabilitation after total knee arthroplasty (TKA) with a rehabilitation training instrument NEO-GAIT. Methods: Sixty patients who received TKA from January 2017 to July 2017 in the Third Hospital of Hebei Medical University were randomly assigned to receive rehabilitation training with continuous passive motion (CPM) or NEO-GAIT with random number (30 cases in CPM group, included 8 males and 22 females; 30 cases in NEO-GAIT group, included 6 males and 24 females). The visual analogue scale (VAS) evaluation of pain, the postoperative range of motion (ROM) of the knee at the 5th and 10th day, and the Hospital for Special Surgery Knee-rating Score (HSS) at 1-month and 3-month follow-up were recorded. The data were compared between the two groups with paired t test. Results: All the patients were followed-up for more than 3 months. The mean VAS in CPM group and NEO-GAIT group on the 5th day was 2.4±1.1, 2.8±1.3, respectively; and it was 2.1±1.1, 2.5±1.2 respectively on the 10th day after the operation (t=-1.618, -1.505, both P>0.05). There was no significant difference in ROM on the 5th day after operation between the 2 groups (84°±12° vs 85°±12°, t=-0.377, P>0.05); however, it was remarkably higher in the NEO-GAIT group (95°±11°) than that in CPM group (88°±8°) on the 10th day after the operation (t=-3.002, P<0.05). The HSS score at 1-month follow-up in CPM group was 72±9, and it was 84±10 in NEO-GAIT group (t=-5.358, P<0.05); but it was comparative between the two groups at the 3-month follow-up (87±5 vs 89±5, t=-1.575, P>0.05). Conclusion: NEO-GAIT plays a more active and effective role in promoting postoperative rehabilitation after TKA than CPM.

[Effect of mesenchymal stem cells on the apoptosis of breast cancer cells induced by cisplatin].

Objective: To investigate the effect of mesenchymal stem cells (MSCs) on apoptosis of breast cancer cell line MCF-7 induced by cisplatin (DDP), MSCs derived from breast cancer (BC-MSCs) or adjacent non-cancerous tissues (BN-MSCs) were isolated, cultured and identified. Methods: BC-MSCs and BN-MSCs were isolated and cultured by tissue adherent method. The differentiation potential of BC-MSCs was detected by osteogenic and adipogenic induction, and cell surface markers of BC-MSCs and BN-MSCs were evaluated by flow cytometry. MCF-7 cells were co-treated with DDP and conditioned medium (CM) collected from BC-MSCs and BN-MSCs after being cultured for 48 hours, respectively. Inhibition rate of cell proliferation was evaluated by MTT. Cell apoptosis and viability were detected by MUSE cell analyzer. Cytokines in MSC-CM were detected by Luminex liquid chip. Interleukin 6 (IL-6) mRNA expressions in MCF-7 cells with different treatment were detected by RT-PCR. Results: The morphology of BC-MSCs and BN-MSCs successfully isolated and cultured was uniform fibroblast-like clusters under the microscope. These cells expressed high levels of CD29 and CD44, but neither CD14 nor CD34 were detected. MSCs could also differentiate into osteoblasts and adipocytes after specific induction. After treatment with 2.5, 5, 10, 20, 40 and 80 µmol/L DDP, the inhibitory rates of proliferation of MCF-7 cells in DDP group were (17.33±2.00)%, (22.37±0.73)%, (30.77±1.23)%, (44.93±1.27)%, (62.03 ±1.97)% and (73.93±1.10)%, respectively. While the inhibitory rates of DDP+ BC-MSCs group were (8.27±0.63)%, (11.50±1.30)%, (20.57±0.93)%, (32.60 ±1.90)%, (52.27±0.73)% and (62.13±2.17)%, respectively. The inhibitory rates of DDP+ BN-MSCs group were (12.90±1.60)%, (16.53±2.87)%, (25.90±1.50)%, (39.40±2.40)%, (57.40±0.70)% and (69.03±1.07)%, respectively. The inhibitory rates of DDP+ BC-MSCs group were significantly lower than those of DDP group (P<0.05). The apoptotic rates of MCF-7 cells in DDP group, DDP+ BC-MSCs group and DDP+ BN-MSCs group were (47.77±1.98)%, (29.20±2.12)% and (37.92±2.21)%, respectively. The apoptotic rates of DDP group was significantly higher than that of DDP+ BC-MSCs group (P<0.05). The cell viabilities of MCF-7 in DDP group, DDP+ BC-MSCs group and DDP+ BN-MSCs group were 0.52±0.02, 0.72±0.02 and 0.64±0.02, respectively. The cell viability of DDP group was significantly lower than that of DDP+ BC-MSCs group (P<0.05). The result of Luminex liquid chip analysis showed that, the level of IL-6 in BC-MSCs group increased 2.50±0.68 fold when compared with BN-MSCs group (P<0.05). The relative expressions of IL-6 mRNA in DDP group and DDP+ BC-MSCs group were 1.02±0.10 and 7.58±0.55, respectively, with a statistically significant difference (P<0.01). The apoptotic rates of MCF-7 cells in DDP+ BC-MSCs group with or without IL-6 neutralizing antibody were (27.41±1.95)% and (42.45±2.87)%, respectively, with a statistically significant difference (P<0.05). The cell viabilities of MCF-7 cells in DDP+ BC-MSCs group with or without IL-6 neutralizing antibody were (72.40±2.60)% and (59.76±3.89)%, respectively, with a statistically significant difference (P<0.05). Conclusions: BC-MSCs and BN-MSCs have been isolated and cultured successfully. Compared with BN-MSCs, BC-MSCs could attenuate the effect of DDP on MCF-7 cells, evidently decrease the apoptosis and increase the proliferation and vitality in an IL-6 dependent manner.

[Value of texture analysis in evaluating liver cancer recurrence after transarterial chemoembolization].

Objective: To investigate the feasibility of contrast-enhanced computer tomography (CT) texture analysis in predicting early recurrence after transarterial chemoembolization (TACE) in patients with liver cancer. Methods: A retrospective analysis was performed for 47 patients with liver cancer confirmed by liver biopsy and digital subtraction angiography who underwent upper abdominal contrast-enhanced CT scan before TACE, and according to the presence or absence of focal recurrence within half a year, these patients were divided into early recurrence (ER) group and non-early recurrence (NER) group. The texture analysis was used to delineate tumor boundary layer by layer on the axial contrast-enhanced CT image before liver cancer surgery, and related parameters of tumor heterogeneity, including entropy, mean, non-uniformity, skewness, and kurtosis, were obtained. The independent samples t-test was used for comparison of texture parameters between the two groups. The receiver operating characteristic (ROC) curve was used for the analysis of entropy, mean, and non-uniformity, and the area under the ROC curve (ROC), optical cut-off value, sensitivity, and specificity were calculated to evaluate the efficiency of texture analysis in predicting early focal recurrence after TACE. Results: There were 20 patients in the ER group and 27 in the NER group. The ER group had a maximum major axis length of 88.2±36.3 mm and a maximum minor axis length of 41.4±21.4 mm, and the NER group had a maximum major axis length of 66.9±30.2 mm and a maximum minor axis length of 29.3±19.8 mm; the ER group had significantly higher maximum major and minor axis lengths than the NER group (t = 4.89 and 4.62, P < 0.001). The ER group had significantly higher entropy and non-uniformity values than the NER group, and there were no significant differences in skewness and kurtosis between the two groups. Entropy, non-uniformity, and mean had high efficiency in predicting early recurrence after TACE, and the optimal cut-off value of entropy was 4.135. Conclusion: Volumetric texture analysis of contrast-enhanced CT images before liver cancer surgery has a high value in predicting early recurrence after TACE.

Proximal femoral nails antirotation, Gamma nails, and dynamic hip screws for fixation of intertrochanteric fractures of femur: A meta-analysis.

BACKGROUND: Which surgical strategy is the best one for intertrochanteric fractures remains a controversial issue. Dynamic hip screw (DHS) and Gamma nail were commonly used but often associated with some complications, such as fixation failure and implant-related fractures. Meanwhile, proximal femoral nail anti-rotation (PFNA) fixation has recently been developed for minimally invasive surgery to reduce the complications rate. To facilitate the clinical decision-making, we conducted an updated meta-analysis to discuss the optimal treatment of intertrochanteric fractures aiming to determine which implant gives the lower rates of blood loss, complications (peri-implant fracture, fixation failure, infection, thromboembolic), reoperation, and mortality, as well as the minimal duration related to surgery (fluoroscopic exposure, surgery and hospital stay). PATIENTS AND METHODS: Seven electronic databases were searched for randomized controlled trials (including OVID, Springer, Google Scholar, PubMed, Cochrane library, Embase, and Web of Science). Fourteen studies with 1983 patients were included. The modified Jadad Scale was used to assess the methodological quality of these studies. Risk of bias in the included studies was assessed using the Cochrane Risk of Bias tool. Comparison among the three groups was based on twelve indicators, including operative time, fluoroscopy time, operative blood loss, length of hospital stays, wound infection or hematoma, pneumonia, thromboembolic complications, fixation failure, operative fracture of femur, later fracture of femur, reoperation, and mortality. RESULTS: (1) PFNA group versus DHS group: PFNA was associated with less blood loss (mean difference (MD) -253.86, 95% CI -270.25 to 237.47; P<0.00001) and lower rate of fixation failure (MD 0.20, 95% CI 0.07 to 0.59; P=0.004), but led to more fluoroscopy time (MD 2.11, 95% CI 1.78 to 2.43; P<0.00001). (2) PFNA group versus Gamma nail group: PFNA led to less blood loss (MD -55.30, 95% CI -60.07 to -50.53; P<0.00001), shorter fluoroscopy time (MD -0.50, 95% CI -0.55 to -0.45; P<0.00001) and length of hospital stay (MD -0.20, 95% CI -0.27 to -0.13; P<0.00001). (3) DHS group versus Gamma nail group: DHS was associated with lower rate of operative fracture of femur (MD 0.31, 95% CI 0.11 to 0.89; P=0.03), later fracture of femur (MD 0.16, 95% CI 0.06 to 0.43; P=0.0004), and reoperation (MD 0.49, 95% CI 0.27 to 0.88; P=0.02), but caused more blood loss (MD 29.49, 95% CI 8.27 to 50.70; P=0.006). In contrast, there was no difference regarding operative time, infection hematoma, pneumonia, thromboembolic events, and mortality. DISCUSSION: PFNA should be a priority choice for treatment of intertrochanteric fractures with minimal rate of fixation failure, less blood loss and shorter length of hospital stay. DHS has distinct advantages over Gamma nail with lower rate of plant-related complications and should be preferred device for intertrochanteric fractures. However, owing to the low quality evidence currently available, more high-quality RCTs are needed to confirm these findings. LEVEL OF EVIDENCE: Level II.

Methylation regulation of liver-specific microRNA-122 expression and its effects on the proliferation and apoptosis of hepatocellular carcinoma cells.

The regulation mechanism and significance of microRNA-122 (miRNA-122) expression are unclear. The aim of this study was to investigate the effects of DNA methylation on liver-specific miRNA-122 expression, cell proliferation, and apoptosis in hepatocellular carcinoma. Methylation of the miRNA-122 promoter region was detected through methylation sequencing. The level of miRNA-122 expression was measured using real-time quantitative polymerase chain reaction. The proliferation and apoptosis of hepatocellular cell lines were detected using flow cytometry and Cell Counting Kit-8 assays. Compared with those in human primary hepatocytes, methylation levels of the miRNA-122 promoter in the Huh7, HepG2, and QSG-7701 cell lines were significantly increased (P = 0.000). Similarly, levels of miRNA-122 expression in these cell lines significantly decreased (P = 0.007). After treatment with 5-aza-2-deoxycytidine, the Huh7 and HepG2 cell lines displayed a significantly lower degree of methylation (P = 0.038 and 0.025), and the levels of miRNA-122 expression were significantly higher (P = 0.008 and 0.003) than those in the blank group. Compared with the blank group, apoptosis of Huh7 and HepG2 cells was significantly increased (P = 0.001 and 0.027). We concluded that the expression of miRNA-122 is regulated by DNA methylation and correlated with apoptosis of liver cancer cells. Methylation regulation of miRNA-122 expression might be involved in the development of hepatocellular carcinoma.

Expression of PI3K/AKT pathway in gastric cancer and its blockade suppresses tumor growth and metastasis.

Phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway plays a crucial role in the formation and progression of many malignancies, and has been shown to be an important therapeutic target for cancer. In the present study, human gastric adenocarcinoma tissues of different grades (N=45) were collected. The protein expression of PI3Kp85α and phosphorylated AKT (p-AKT) was evaluated immunohistochemically in the biopsy samples. PI3K/AKT pathway was blocked by constructed recombinant small hairpin RNA adenovirus vector rAd5-PI3Kp85α (rAd5-P) used to transfect into human gastric cancer SGC-7901cell line. The transfection efficiency of rAd5-P in SGC-7901 cells was observed under fluorescent microscope. The expression of PI3Kp85α, p-AKT, Ki-67 and matrix metallopeptidase-2 (MMP-2) was detected by real-time PCR and Western blot assays. Cell proliferative activities and metastatic capabilities were determined by MTT and Transwell assays. As a consequence, the protein expression of PI3Kp85α and p-AKT was respectively observed in 80.0% and 82.2% gastric adenocarcinoma tissues, elevating with the ascending order of tumor malignancy. Targeted blockade of PI3K pathway decreased the expression of PI3Kp85α, p-AKT, Ki-67 and MMP-2, and inhibited the proliferative activities and metastatic capabilities of gastric cancer cells. In conclusion, PI3Kp85α and p-AKT were strongly expressed in gastric adenocarcinoma tissues, and targeted blockade of PI3K pathway may inhibit gastric cancer growth and metastasis through down-regulation of Ki-67 and MMP-2 expression. PI3K/AKT pathway may represent an important therapeutic target for gastric cancer.

Propofol attenuates hypoxia-induced apoptosis in alveolar epithelial type II cells through down-regulating hypoxia-inducible factor-1α.

To investigate the protective effect of propofol against hypoxia-induced apoptosis in alveolar epithelial type II (ATII) cells and to explore whether hypoxia-inducible factor-1α (HIF-1α) is involved in this process. Primary cultured rat ATII cells were randomly assigned to one of the following four groups, namely, Group C: treated under normoxia (21% O(2)), Group P(20): treated with propofol (20 µM) under normoxia (21% O(2)), Group H: treated under hypoxia (5% O(2)), and Group P(20)-H: pre-treated with propofol (20 µM) before hypoxia exposure (5% O(2)). Apoptosis in ATII cells was detected by Annexin V-FITC binding using FACScan. Expressions of HIF-1α and Bnip3L mRNA and protein in ATII cells were examined by quantitative real-time polymerase chain reaction and Western blotting analysis, respectively. Hypoxia exposure (Group H) significantly increased HIF-1α protein expression (P<0.01 vs. Group C) and significantly promoted apoptosis in ATII cells (P<0.01 vs. Group C). Expression of Bnip3L, a target gene of HIF-1α, was also significantly increased at both mRNA and protein levels in response to hypoxia (P<0.01 vs. Group C). Pretreatment with propofol (20 µM, Group P(20)-H) significantly decreased HIF-1α protein expression (P<0.01 vs. Group H) and significantly inhibited apoptosis in ATII cells (P<0.01 vs. Group H), accompanied by decreased expression of Bnip3L at both mRNA and protein levels (P<0.01 vs. Group H). Propofol (20 µM) can attenuate hypoxia-induced apoptosis in ATII cells and inhibit HIF-1α-hypoxia responsive element (HRE) axis involving Bnip3L, which may partly mediate the cytoprotective effects of propofol.

p53 protein expression and genetic mutation in two primary cell types in pulmonary sclerosing haemangioma.

AIMS: To investigate the significance of p53 protein expression and genetic mutations in two primary cell types in pulmonary sclerosing haemangioma (PSH). METHODS: p53 protein expression in polygonal cells and cuboidal cells in 19 patients with PSH was detected using immunohistochemistry. The two major cell types were captured using laser capture microdissection technology. Mutations in the p53 gene (exons 5-8) were examined using single-stranded conformation polymorphism and DNA sequencing analysis. RESULTS: p53 protein expression and gene mutations were observed in 15.8% (3/19) of cases. In these cases, p53 protein was expressed in the nucleus of both cell types, with higher expression levels and mutation rates in polygonal cells than in surface cuboidal cells. Two cases showed mutation only in the polygonal cells, while one case showed double (separate) mutations in both the polygonal and cuboidal cells. CONCLUSIONS: p53 mutation was exhibited in PSH. The mutation rate in polygonal cells was higher than that in surface cuboidal cells.

Abnormal expression and clinicopathologic significance of p120-catenin in lung cancer.

The aim of this study was to investigate the relationship between the expression of p120ctn in human lung squamous cell carcinoma, adenocarcinoma and its clinicopathologic significance. The expression of p120ctn in tumors and adjacent normal lung tissues from 143 patients was examined by immunohistochemistry and Western blot. Expression of p120ctn occurs mainly in the cell membrane of normal bronchial mucosa. Abnormal expression of p120ctn, including cytoplasmic and reduced membranous expression, was found in 114 of 143 specimens (79.7%) and was significantly associated with poor differentiation, high TNM stage, and lymph node metastasis (P<0.05 for each) but not with histologic subtype. The Kaplan-Meier survival test revealed that abnormal expression of p120ctn was related to poor survival (P<0.001). A Cox regression analysis revealed that abnormal p120ctn expression was an independent factor in predicting patient survival (P=0.024). Compared with that in normal lung tissues, membranous protein level was lower in tumors (P=0.003). Abnormal expression of p120ctn is associated with tumor progression and poor prognosis in lung squamous cell carcinoma and adenocarcinoma. Reduced expression or even the absence of p120ctn isoform 1 and 3 in tumor cell membranes may be responsible for the abnormal expression of p120ctn that has been found in lung cancer.