An efficient TILLING platform for cultivated tobacco.

Targeting-induced local lesions in genomes (TILLING) is a powerful reverse-genetics tool that enables high-throughput screening of genomic variations in plants. Although TILLING has been developed for many diploid plants, the technology has been used in very few polyploid species due to their genomic complexity. Here, we established an efficient capillary electrophoresis-based TILLING platform for allotetraploid cultivated tobacco (Nicotiana tabacum L.) using an ethyl methanesulfonate (EMS)-mutagenized population of 1,536 individuals. We optimized the procedures for endonuclease preparation, leaf tissue sampling, DNA extraction, normalization, pooling, PCR amplification, heteroduplex formation, and capillary electrophoresis. In a test screen using seven target genes with eight PCR fragments, we obtained 118 mutants. The mutation density was estimated to be approximately one mutation per 106 kb on average. Phenotypic analyses showed that mutations in two heavy metal transporter genes, HMA2S and HMA4T, led to reduced accumulation of cadmium and zinc, which was confirmed independently using CRISPR/Cas9 to generate knockout mutants. Our results demonstrate that this powerful TILLING platform (available at http://www.croptilling.org) can be used in tobacco to facilitate functional genomics applications.

The Prevalence of Suicide Attempts and Independent Demographic and Clinical Correlates among Chronic Schizophrenia Patients in Agricultural Areas of China.

This study investigated the lifetime prevalence of suicide attempts (SA) and independent demographic and clinical correlates in stabilized schizophrenia inpatients. A cross-sectional study was conducted in three psychiatric hospitals in Anhui province, an agricultural province located in east China. Psychopathology and depressive symptoms were assessed using the Positive and Negative Syndrome Scale (PANSS) and Hamilton Depression Rating Scale (HAMD), respectively. A total of 315 stable schizophrenia inpatients were interviewed prior to discharge. The lifetime prevalence of SA was 22.2%. Multiple logistic regression analysis revealed that female gender (P < 0.001, OR = 3.4, 95%CI: 1.9-6.0), being married (P = 0.02, OR = 2.2, 95%CI: 1.1-4.4) and having more severe depressive symptoms (P = 0.014, OR = 1.2, 95%CI: 1.01-1.3) were independently and significantly associated with higher risk of SA. Lifetime SA is common among hospitalized schizophrenia patients living in agricultural areas of China. For suicide prevention, regular assessments, appropriate interventions and clinical management should be integrated into a community-based psychiatric service model for this population.

A study of somatization symptoms and low-frequency amplitude fluctuations of emotional memory in adolescent depression.

Studies have revealed that somatization symptoms are associated with emotional memory in adolescents with depressive disorders. This study investigated somatization symptoms and emotional memory among adolescents with depressive disorders using low-frequency amplitude fluctuations (ALFF). Participants were categorized into the somatization symptoms (FSS) group, non-FSS group and healthy control group (HC). The correctness of negative picture re-recognition was higher in the FFS and HC group than in the non-FSS group. The right superior occipital gyrus and right inferior temporal gyrus were significantly larger in the FSS group than those in the non-FSS and HC groups. Additionally, the ALFF in the superior occipital and inferior temporal gyrus were positively correlated with CSI score. Furthermore, the ALFF values in the temporal region positively correlated with correct negative image re-recognition. The negative image re-recognition rate was positively correlated with the ALFF in the left and right middle occipital gyri. These findings indicated that somatization symptoms in adolescent depression are associated with the superior occipital gyrus and inferior temporal gyrus. Notably, somatization symptoms play a role in memory bias within depressive disorders, with middle occipital and inferior temporal gyri potentially serving as significant brain regions.

[Clinical outcomes and bone resection analysis of unilateral double-channel endoscopic technique in treating lumbar disc herniation].

OBJECTIVE: To explore clinical outcomes and bone resection of interlaminar fenestration decompression and unilateral biportal endoscopic (UBE) technique in treating lumbar disc herniation(LDH). METHODS: A retrospective study was performed on 105 patients with single-level LDH treated from December 2019 to December 2021. Fifty-four patients in UBE group,including 32 males and 22 females,aged from 18 to 50 years old with an average of(38.7±9.3) years old,were treated with UBE,29 patients with L4,5 and 25 patients with L5S1. There were 51 patients in small fenestration group,including 27 males and 24 females,aged from 18 to 50 years old with an average of (39.9±10.0) years old,were treated with small fenestration,25 patients with L4,5 and 26 patients with L5S1. Perioperative indexes,such as operation time,postoperative time of getting out of bed and hospital stay were observed and compared between two groups. Visual analogue scale (VAS) and Oswestry disability index (ODI) were compared between two groups before operation and 1,3,6 and 12 months after operation,respectively;and modified MacNab evaluation criteria was used to evaluate clinical efficacy. Amount of bone resection and retention rate of inferior articular process laminoid complex were compared between two groups. RESULTS: All 105 patients were successfully completed operation. Both of two groups were followed up from 6 to 12 months with an average of (10.69±2.49) months. Operation time,postoperative time of getting out of bed and hospital stay were (58.20±5.54) min,(2.40±0.57) d and (3.80±0.61) d in UBE group,and (62.90±7.14) min,(4.40±0.64) d and (4.40±0.64) d in small fenestrum group,respectively;and had statistically difference between two groups(P<0.05). Postoperative VAS of low back and leg pain and ODI in both groups were significantly lower than those before surgery (P<0.05). VAS of lumbar pain in UBE group (1.37±0.49) score was lower than that of small fenestration group (2.45±0.64) score,and had statistically difference (t=9.745,P<0.05). Postoperative ODI in UBE group at 1 and 3 months were (28.54±3.31) % and (22.87±3.23) %,respectively,which were lower than those in small fenestra group (36.31±9.08) % and (29.90±8.36) %,and the difference was statistically significant (P<0.05). There were no significant difference in VAS and ODI between two groups at other time points (P>0.05). According to the modified MacNab evaluation criteria at the latest follow-up,49 patients got excellent result,3 good,and 2 fair in UBE group. In small fenestration group,35 patients got excellent,12 good,and 4 fair. In UBE group,amount of bone resection on L4,5 segment was (0.45±0.08) cm3 and (0.31±0.08) cm3 on the segment of L5S1. In small fenestration group,amount of bone resection on L4,5 segment was (0.57±0.07) cm3 and (0.49±0.04) cm3 on the segment of L5S1,and amount of bone resection of lower articular process laminar complex on the same segment in UBE group was less than that in small fenestration group (P<0.05). In UBE group,retention rate of laminoid complex on L4,5 segment was (0.73±0.04) and L5S1 segment was (0.83±0.03),while L4,5 segment was(0.68±0.06) and L5S1 segment was (0.74±0.04) in small fenestration group,the lower articular process laminar complex retention rate in UBE group was higher than that in small fenestration group(P<0.05). CONCLUSION: Both unilateral double-channel endoscopy and small fenestration of laminae could achieve good clinical results in treating LDH,but UBE has advantages of less trauma,higher efficiency,faster postoperative recovery and less damage to bone structure.

[CT manifestations of liver involvement in children with Langerhans cell histiocytosis].

OBJECTIVE: To investigate the computed tomography (CT) manifestations of liver involvement in children with Langerhans cell histiocytosis (LCH). METHODS: Retrospective analysis was performed on 9 LCH children with liver involvement confirmed by clinical, laboratory and pathological examinations to investigate the CT manifestations of this condition. These children, including 6 males and 3 females, had undergone both plain CT scan and dual-phase (the arterial and portal venous phases) contrast-enhanced CT scan. RESULTS: The main CT manifestations included hepatomegaly (8 cases); periportal dendritic hypodense lesions or "periportal halo sign" (7 cases) which were mildly or moderately enhanced in the arterial phase; intrahepatic bile duct dilatation (5 cases); lymphadenopathy in the hepatic hilar or retroperitoneal region (4 cases); and diffuse small hypodense nodules (3 cases), which showed annular enhancement on the contrast-enhanced CT scan. CONCLUSIONS: CT findings may be helpful in the early diagnosis and treatment of LCH in children.

[Application Value of Plasma vWF and ADAMTS-13 in Evaluating Postoprative Vascular Endothelial Injury and Prognosis in Children with VSD].

OBJECTIVE: To dynamically observe the levels and activities of von Willebrand factor (vWF) and ADAMTS-13 (a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13) in plasma of children with congenital ventricular septal defect (VSD) during perioperative period, and explore the value of plasma vWF antigen (vWF:Ag) and ADAMTS-13 activity (ADAMTS-13: AC) in evaluating vascular endothelial injury and prognosis in children with VSD. METHODS: In this cross-sectional study, a total of 74 children with VSD who underwent surgical treatment in TEDA International Cardiovascular Hospital from September 2018 to March 2019 were enrolled in the observation group. Among them, there were 28 cases of pure VSD, 32 cases of VSD combined with pulmonary hypertension, and 14 cases of VSD combined with valvular heart disease. 31 healthy children who underwent physical examination in Tianjin Children's Hospital during the same period were collected as the control group. The biochemical indexes of the children at admission were recorded. Peripheral plasma was collected at admission, postsurgery day 0 and day 1, respectively, and the levels of vWF activity (vWF:AC), vWF:Ag, ADAMTS-13 antigen (ADAMTS-13:Ag) and ADAMTS-13:AC were detected. RESULTS: The level of plasma vWF:Ag and vWF:AC in the observation group before surgery were significantly lower than those in the control group (P<0.001), and increased continuously, on postsurgery day 0 and day 1 (P<0.001). The level of ADAMTS-13:Ag in the observation group before surgery was significantly higher than that in the control group (P<0.001), which decreased significantly on postsurgery day 0 (P<0.001), and increased significantly on postsurgery day 1 compared with postsurgery day 0 (P=0.033). The level of ADAMTS-13:AC in the observation group before surgery was significantly lower than that in the control group (P=0.015), which decreased significantly on postsurgery day 0 (P=0.037), and increased on postsurgery day 1, but the difference was not statistically significant (P=0.051). The changes of vWF and ADAMTS-13 in the three subgroups were basically similar to the observation group. vWF: Ag/ADAMTS-13: AC ratio on postsurgery day 0 and day 1 had high diagnostic value in vascular endothelial injury (AUC=0.80, P<0.001; AUC=0.93, P<0.001). Preoperative vWF and ADAMTS-13 levels, and related baseline indicators were not correlated with postoperative infection, bleeding, thrombosis,etc. CONCLUSION: Preoperative vWF: Ag, vWF: AC and ADAMTS-13: AC levels in children with VSD are low, while the level of ADAMTS-13: Ag is high. After surgery, the levels of vWF: Ag and vWF: AC are increased and the level of ADAMTS-13: Ag is decreased. The postoperative vWF: Ag/ADAMTS-13: AC ratio shows high diagnostic value in evaluating vascular endothelial injury. There is no correlation between preoperative vWF and ADAMTS-13 levels with perioperative clinical events.

[Clinical implication and association between local and systemic levels of interleukin-1ß and interleukin-10 in patients with coronary artery disease].

OBJECTIVE: To compare the systemic and local near atherosclerosis lesion levels of pro-inflammatory factor interleukin-1ß (IL-1ß) and anti-inflammatory factor IL-10 in patients with coronary artery disease (CAD). METHODS: Plasma samples were collected from 30 individuals without angiographical coronary artery stenosis (control group), 90 patients with CAD (stable angina pectoris, SA, n = 30, unstable angina pectoris/non-ST-segment elevation myocardial infarction, UA/NSTEMI, n = 30 and ST-segment elevation myocardial infarction, STEMI, n = 30). During diagnostic coronary angiography or interventional procedures, systemic samples were obtained from aorta root in all patients (n = 120), local samples from distal of the coronary lesion in patients with CAD (n = 90), and samples from coronary sinus of 14 patients with STEMI. IL-1ß and IL-10 were determined by ELISA method. RESULTS: The result showed systemic levels of IL-1ß were lg(-1) (0.97 ± 0.42), lg(-1) (0.98 ± 0.43), lg(-1) (1.21 ± 0.42), lg(-1) (1.30 ± 0.43) ng/L in the control, SA, UA/NSTEMI and STEMI groups, were significantly higher in UA/NSTEMI and STEMI groups compared with the control group (P < 0.05, P < 0.01); systemic IL-10 levels were lg(-1) (0.77 ± 0.29), lg(-1) (0.73 ± 0.45), lg(-1) (0.75 ± 0.35), lg(-1) (1.14 ± 0.36) ng/L in the four groups and was significantly higher in STEMI group than the control group (P < 0.01). The local concentration of IL-1ß and IL-10 were similar as the systemic levels in SA group [lg(-1) (0.98 ± 0.41), lg(-1) (0.67 ± 0.47) ng/L], local IL-1ß [lg(-1) (1.22 ± 0.48) ng/L] was similar while local IL-10 [lg(-1) (0.89 ± 0.46) ng/L] was significantly higher than the systemic levels in UA/NSTEMI group. The local levels of IL-1ß and IL-10 [lg(-1) (1.45 ± 0.45), lg(-1) (1.35 ± 0.31) ng/L] were both significantly higher than the systemic levels in STEMI group (all P < 0.01). The IL-1ß levels of systemic, local and coronary sinus in STEMI patients with acute totally occluded left coronary artery [lg(-1) (1.47 ± 0.37), lg(-1) (1.65 ± 0.34), lg(-1) (1.53 ± 0.35)ng/L] and the IL-10 levels [lg(-1) (1.06 ± 0.48), lg(-1) (1.34 ± 0.39), lg(-1) (1.34 ± 0.23) ng/L] were similar. The level of IL-1ß in coronary sinus was significantly lower than in culprit lesion (P < 0.05) while IL-10 levels were similar at these two sites (P > 0.05). CONCLUSION: The systemic level of pro-inflammatory marker IL-1ß and anti-inflammatory marker IL-10 could not reliably reflect the local inflammatory status near the atherosclerosis plaque locations.

[Efficiency and safety of thrombus aspiration plus intra-infarct-related artery administration of tirofiban during primary angioplasty].

OBJECTIVE: To investigate whether thrombus aspiration plus intra-infarct-related artery bolus administration of tirofiban via the aspiration catheter is superior to thrombus aspiration alone in improving myocardial perfusion in patients with ST-elevation myocardial infarction (STEMI) undergoing primary angioplasty. METHODS: In this single center retrospective study, 108 patients with STEMI who underwent angioplasty after thrombus aspiration plus intra-infarction related artery 500 µg tirofiban administration, with subsequent 12-hour intravenous infusion of 0.1 µg×kg(-1)×min(-1) after angioplasty (thrombus aspiration + tirofiban group) and 108 matched control patients with STEMI who underwent angioplasty after thrombus aspiration (thrombus aspiration group). The primary end points included thrombolysis in myocardial infarction (TIMI) flow immediately after angioplasty, complete ST-segment elevation resolution (> 70%) at 90 minutes after angioplasty and the peak of creatine kinase-MB (CK-MB) and troponin I (TnI). The secondary end points were the left ventricular ejection fraction (LVEF) in the hospital and at 9 months follow-up as well as major adverse cardiac events (MACE: cardiac death, target vessel revascularization, re-infarction) at 9 months and any bleeding events. RESULTS: Baseline characteristics of the two groups were well-balanced. The TIMI 3 flow rate (97.22% vs. 87.04%, P = 0.011) and the complete ST-segment resolution rate (66.67% vs. 50.91%, χ(2) = 6.129, P = 0.047)were significantly higher in the thrombus aspiration + tirofiban group than in the thrombus aspiration group. The peak of CK-MB (83.9 U/L vs. 126.1 U/L, P = 0.034) and TnI (42.7 ng/ml vs. 72.5 ng/ml, P = 0.029) were significantly lower in the thrombus aspiration + tirofiban group than in the thrombus aspiration group. LVEF in the hospital favored thrombus aspiration + tirofiban the group (45.7% ± 10.8%, 42.9% ± 9.9%, t = 1.99, P = 0.049). There was a tendency to decreased MACE rate at 9-month follow-up, which favored thrombus aspiration + tirofiban the group (logrank χ(2) = 2.865, P = 0.09). Bleeding events were similar between the two groups. CONCLUSION: Thrombus aspiration plus intra-infarct-related artery bolus administration of tirofiban in patients with STEMI undergoing primary angioplasty may improve myocardium perfusion, attenuate myocardial ischemia and result in a better clinical prognosis compared to thrombus aspiration alone.

[Pathological analysis of aspirated materials from the culprit lesion in patients with acute myocardial infarction].

OBJECTIVE: To analyze the components of retrieved materials from the culprit lesion in ST-segment elevation myocardial infarction (STEMI) patients by manual aspiration during primary percutaneous coronary intervention (PCI). METHODS: Visible retrieved materials were collected, fixed in formalin and processed for paraffin embedding, sectioned and stained with hematoxylin and eosin (HE). The retrieved materials were microscopically divided into erythrocyte-rich thrombi, platelet/fibrin-rich thrombi, combined thrombi (similar proportions of erythrocytes and platelet/fibrin components), atherosclerotic plaque materials and edematous components. Based on pathological findings, thrombus materials were classified into fresh (< 1 d), lytic (1-5 d), fresh/lytic and organized thrombi (> 5 d) after formation. All patients were further classified into plaque positive and plaque negative groups. Clinical and angiographic data were also obtained for analyzing possible association between pathological findings and surrogates of myocardial reperfusion, including myocardial blush grade (MBG), enzymatic estimated infarction size (peak CK and CK-MB levels), left ventricular end diastolic diameter (LVEDD) and left ventricular ejection fraction (LVEF) which were assessed 16 h after procedure. RESULTS: Visible samples were collected from 49 patients by manual catheter aspiration (thrombus components in 46 patients, atherosclerotic plaque only in 3 patients). Frequency of erythrocyte-rich thrombi, platelet/fibrin-rich thrombi and combined thrombi were 41.3% (19/46), 30.4% (14/46) and 28.2% (13/46), respectively. The incidence of fresh, lytic, fresh/lytic and organized thrombi were 47.8% (22/46), 32.6% (15/46), 10.9% (5/46) and 8.7% (4/46), respectively. Plaque materials were found in 57.1% (28/49) patients, including ruptured plaque accompanied by thrombus formation [8.2% (4/49)], fibrous plaque [6.1% (3/49)] and thickened intima [2.0% (1/49)]. Baseline characteristics did not differ between plaque positive (n = 28) and plaque negative (n = 21) groups. Ratios of MBG 3 were higher in plaque positive group than in plaque negative group [82.1% (23/28) vs. 52.4% (11/21), P = 0.025]. Peak CK and CK-MB levels were lower in the former than in the later [(1705 +/- 1647) U/L vs. (2629 +/- 2013) U/L, P = 0.042; (146 +/- 136) microg/L vs. (258 +/- 215) microg/L, P = 0.016; respectively]. Furthermore, LVEF were higher in plaque positive group than in plaque negative group (0.59 +/- 0.10 vs. 0.52 +/- 0.08, P = 0.012). CONCLUSION: Manual catheter aspiration during primary PCI in STEMI patients is an effective way for removing thrombus and plaque materials, and plaque debulking before stenting or pre-dilation and this procedure might probably improve myocardial reperfusion, limit infarction size and improve cardiac function.

Predictors for New Native-Vessel Occlusion in Patients with Prior Coronary Bypass Surgery: A Single-Center Retrospective Research.

OBJECTIVES: Chronic total occlusion (CTO) is prevalent in patients with prior coronary artery bypass grafting (CABG). However, data available concerning the prevalence of new-onset CTO of native vessels in patients with prior CABG is limited. Therefore, the objective of the study is to determine predictors for new native-vessel occlusion in patients with prior coronary bypass surgery. METHODS: 354 patients with prior CABG receiving follow-up angiography are selected and analyzed in the present study, with clinical and angiographic variables being analyzed by logistic regression to determine the predictors of new native-vessel occlusion. RESULTS: The overall new occlusion rate was 35.59%, with multiple CTOs (42.06%) being the most prevalent (LAD 24.60% and RCA 18.25%, respectively). Additionally, current smoking (OR: 2.67; 95% CI: 2.60 to 2.74; p=0.01), reduced ejection fraction (OR: 1.76; 95% CI: 1.04 to 2.97; p=0.04), severe stenosis (OR: 3.65; 95% CI: 2.55 to 5.24; p=0.01), and diabetes mellitus (OR: 1.86; 95% CI: 1.34 to 2.97; p=0.04) serve as the independent predictors for new native-vessel occlusion. CONCLUSION: As to high incidence of postoperative CTO, appropriate revascularization strategies and postoperative management should be taken into careful consideration.

Olanzapine Versus Risperidone in Children and Adolescents with Psychosis: A Meta-Analysis of Randomized Controlled Trials.

OBJECTIVE: To compare the efficacy and safety of olanzapine and risperidone in children and adolescents (aged ≤18 years) with psychosis by conducting a meta-analysis of randomized controlled trials (RCTs). METHODS: Several English and Chinese databases were searched for studies published before February 8th, 2017. Two independent investigators screened the studies according to prespecified criteria and extracted the data. Review Manager 5.3 was used to conduct the data synthesis. RESULTS: Eight RCTs involving 457 participants (225 participants in the olanzapine group and 232 participants in the risperidone group) were included. No significant differences were observed in the mean scores on the Positive and Negative Syndrome Scale/Brief Psychiatric Rating Scale (standard mean difference [SMD] = -0.06, 95% confidence intervals [CI] = [-0.31, 0.19], p = 0.63), the positive symptom scores (SMD = -0.09, 95% CI = [-0.32, 0.15], p = 0.48), or the negative symptom scores (SMD = -0.11 95% CI = [-0.34, 0.13], p = 0.38) between the two groups. Regarding adverse effects, the mean increases in weight (MD = 2.90, 95% CI = [1.41, 4.39], p = 0.0001), body mass index (MD = 0.90, 95% CI = [0.42, 1.38], p = 0.0003), and incidence of hypersomnia (risk ratios [RR] = 1.98, 95% CI = [1.15, 3.43], p = 0.01) were higher in the olanzapine group, while the incidence of insomnia (RR = 0.31, 95% CI = [0.11, 0.85], p = 0.02), prolactin elevation (RR = 0.11, 95% CI = [0.01, 0.85], p = 0.03), myotonia (RR = 0.12, 95% CI = [0.03, 0.49], p = 0.003), tremor (RR = 0.22, 95% CI = [0.08, 0.63], p = 0.005), and akathisia (RR = 0.27, 95% CI = [0.12, 0.57], p = 0.0007) was higher in the risperidone group. CONCLUSIONS: There is no significant difference in efficacy between olanzapine and risperidone for the treatment of children and adolescents with psychosis, but the side effect profiles of these two medications differ. High-quality RCTs are needed before recommending clinical treatment in children and adolescents.

[Oral ferric ammonium citrate solution in MRCP for diagnosing low-level obstructive jaundice].

OBJECTIVE: To evaluate the value of oral ferric ammonium citrate solution as a gastrointestinal contrast agent in diagnosing low-level obstructive jaundice. METHODS: Thirty-six patients who were suspected of low-level obstructive jaundice were performed with magnetic resonance cholangiopancreatography (MRCP) and conventional MRI before and after the administration of oral ferric ammonium citrate solution. The diagnostic accuracy for evaluating the site and the cause of obstruction was compared with other diagnostic modalities. RESULTS: The image qualities of single-slice and multi-slice MRCP were improved markedly. The accuracy of MRCP for evaluating the site of obstruction was 97.22%, which was superior to US (P<0.05). There were no significant differences among the MRCP, CT, and ERCP. CONCLUSION: Oral ferric ammonium citrate solution can significantly improve the image quality of MRCP. FAC-MRCP is a simple, safe, and noninvasive technique with excellent accuracy in the diagnosis of low-level obstructive jaundice.

[Expression of Ang2 and Tie2 and their relation with the angiogenesis of hepatocellular carcinoma in rats].

OBJECTIVE: To investigate the relationship between the expression of Ang2, Tie2 and the angiogenesis of hepatocellular carcinoma in rats. METHODS: Thirty-eight healthy male rats were randomly divided into 3 groups: 5 rats in the control group; 25 rats in the experimental group were equally divided into 5-day, 10-day, 15-day, 20-day, and 25-day groups; the other 8 rats were used as the supplement of the experimental group. An allogenic transplanted rat model of CBRH-7919 hepatocellular carcinoma in situ was established by immunosuppression. The expressions of Ang2 and Tie2 were detected by immunohistochemical staining in cancerous tissues of different developmental stages and liver tissues of the control group. At the same time, microvessel density was determined by anti-CD31 immunohistochemical staining. RESULTS: CBRH-7919 hepatocellular carcinoma models were successfully set up in 24 rats. The expression level of Ang2 and Tie2 in cancerous tissues was much higher than that of liver tissues of the control group (P <0.05). The overexpression of Ang2 was pristine and continuous in different developmental stages. The expressions of Ang2 and Tie2 positively correlated with microvessal density in hepatocellular carcinoma (P<0.05). CONCLUSION: The up-regulation of Ang2 and Tie2 may play important roles in the angiogenesis of hepatocellular carcinoma. Ang2 may participate in the start of angiogenesis of hepatocellular carcinoma.

Six generations of epidermolytic palmoplantar keratoderma, associated with a KRT9 R163W mutation.

Epidermolytic palmoplantar keratoderma (EPPK) is a rare autosomal dominant skin disorder characterized by diffuse hyperkeratosis on the palms and soles. Whole-exome sequencing (WES) has become a powerful tool for the detection of rare causal variants of Mendelian disorders. However, no causal gene for EPPK in the Uygur population has been identified until now, and no treatment exists than can address the underlying pathology.WES analysis was undertaken on two individuals from a large Uygur EPPK pedigree whose disease locus mapped to 17q21.2 (chr:38994621-39893408) following previous linkage analysis. KRT9 (NM_000226.3:c.487C>T, p.Arg163Trp), and KRT15 (XM_005257346.1:c.212G>T, XP_005257403.1:p.Gly71Val) located in this region, have been identified as two candidate causative genes for EPPK in the Uygur family. Sanger sequencing was conducted on this region in other affected individuals (n = 38) from this family, non-affected individuals (n = 56) from this family and 100 unrelated controls. The missense mutation KRT9 c.487C>T, identified in this large Uygur population, is a potential causative mutation. To date, EPPK has no effective therapy, and siRNA is a potential avenue for EPPK therapy. To investigate this, full-length wild-type Keratin9 (KRT9; pKRT9-WT) and p.Arg163Trp (pKRT9-R163W) were then transfected into HaCaT cells. The small interfering RNAs targeting the KRT9 R163W mutant and wildtype KRT9 were transfected into HaCaT cells, and total RNA isolated at 72 h post-transfection. Quantitative polymerase chain reaction and western blotting were used to analyse the effects of knock-down on KRT9 mRNA and protein levels, respectively. siRNA was shown to specifically inhibit mutant KRT9 mRNA and protein expression (p < 0.01, with 95% confidence limits). Our study suggests that KRT9 is a causal gene for EPPK. This information is helpful for understanding the pathogenesis of EPPK in the Uygur population and raises the possibility of designing a novel siRNA treatment strategy for this population of EPPK patients.

Simultaneous silencing of XIAP and survivin causes partial mesenchymal-epithelial transition of human pancreatic cancer cells via the PTEN/PI3K/Akt pathway.

Pancreatic cancer has one of the highest mortality rates among malignant tumors and is characterized by rapid invasion, early metastasis and chemoresistance. X-linked inhibitor of apoptosis (XIAP) and survivin are two of the most important members of the IAP family. Previous studies have shown that XIAP and survivin were overexpressed in pancreatic cancer and were closely associated with cell proliferation and chemoresistance to gemcitabine. In the present study, stable inhibition of XIAP and survivin in Panc-1 cells was performed using lentivirus-carried short hairpin RNAs. The expression of XIAP, survivin, E-cadherin, Slug, phosphatase and tensin homolog (PTEN) and phosphorylated Akt was then measured. In addition, cell proliferation, apoptosis, invasion and migration were assessed. The results showed that stable inhibition of XIAP and survivin expression in Panc-1 cells significantly reduced cell proliferation, increased apoptosis and partially reversed the epithelial-mesenchymal transition (EMT). Furthermore, the results of the present study demonstrated that the partial reversal of the EMT was accompanied by inhibited cell invasion and migration as well as increased chemosensitivity to gemcitabine in pancreatic cancer cells; this was indicated to be mediated via the PTEN/phosphatidylinositol 3-kinase/Akt signaling pathway. In conclusion, these results suggested that simultaneous inhibition of XIAP and survivin may be a promising strategy for the treatment of pancreatic cancer.

A rare case of extra-adrenal pheochromocytoma localized to the ovary and detected via abdominal computed tomography angiography.

Extra-adrenal pheochromocytomas are rare tumors that originate from the chromaffin tissue of the sympathetic nervous system. Ovarian extra-adrenal pheochromocytoma is even more rare. The present study reports a rare case of an extra-adrenal pheochromocytoma that was localized to the right ovary, but was gynecologically asymptomatic. Computed tomography angiography (CTA) detected the tumor and indicated that it was well defined, highly vascularized and obtained its blood supply from the right ovarian artery. This is the second case of ovarian extra-adrenal pheochromocytoma reported in the literature, and the first description of the CTA manifestations in the ovary. Gynecologists and radiologists should consider the possibility that an ovarian mass could be an extra-adrenal pheochromocytoma, which would allow time to prepare appropriately for the surgical removal of the mass.

HIV DNA and immune alteration during successful HAART.

BACKGROUND: Highly Active Antiretroviral therapy (HAART) can effectively reduce the viral load to undetectable levels in most HIV-infected patients. However, some patients may still experience impaired immunologic response associated with increased risk of disease progression and death. OBJECTIVE: The objective of this study was to assess the impact of the HIV DNA load on the immune alteration during successful HAART. METHODS: 40 chronic HIV-infected adults initiating HAART were followed for 24 months. The CD4 count, HIV viral load, HIV DNA load, and levels of γ-cytokines IL-2, IL-7 and IL-21 were monitored at baseline (month 0), month 6, 12, 18 and 24 following HAART initiation. RESULTS: The plasma viral load decreased significantly and remained below the detection limits after six months treatment. Likely the HIV DNA load decreased significantly in both cells during 12 months, was undetectable in CD14 monocytes after 18 months, but remained higher in CD3+ T cells during all the follow up. In addition, the HIV DNA load correlated positively between T cells and monocytes in 10 patients who maintained higher HIV DNA load in both cells during 12 months. The CD4 count, IL-2, and IL-21 levels increased significantly during 12 months, whereas IL-7 decreased significantly during 18 months, regardless of the HIV DNA load in T cells. Patients with CD4 count below 200/µl maintained higher HIV DNA load and showed lower increase in CD4 count compared to patients with CD4 count above 200/µl. In patients showing undetectable HIV DNA load in both cells, neither IL-2 nor IL-21 correlated with the CD4 count even after 24 months despite their partial restoration. CONCLUSION: These results suggest that the HIV DNA load could continuously hamper the CD4 restoration and γ-cytokines functional activities during HAART. This action seemed to be more severe in patients with pre-HAART CD4 count below 200/µl. The CD14 monocytes may contribute to this action as source of T cell infection both before and during HAART.

Comparison of safety and efficacy between fondaparinux and nadroparin in non-ST elevation acute coronary syndromes.

BACKGROUND: American College of Cardiology/American Heart Association/European Society of Cardiology (ACC/AHA/ESC) guidelines gave fondaparinux a class I recommendation for use in patients with non-ST elevation acute coronary syndromes (NSTE-ACS) undergoing invasive or conservative strategy. Nadroparin is one of the common anticoagulants used in NSTE-ACS in China. Accordingly, this study compared the safety and efficacy between fondaparinux and nadroparin in patients with NSTE-ACS. METHODS: In this prospective, randomized, open-label, and single center study, a total of 300 patients with NSTE-ACS were randomized to receive either fondaparinux (group F, n = 150, 2.5 mg/d) or nadroparin (group N, n = 150, 0.1 ml/10 kg q12 h) for a mean of 4 days. The primary safety endpoint was the incidence of major or minor bleeding at 9 days that was not related to coronary artery bypass grafting (CABG). The primary efficacy endpoints included death, myocardial infarction, or recurrent ischemia at 9 days. All patients underwent a 180-day follow-up. RESULTS: Baseline characteristics were well matched between the two groups. There was a non-significant 28% relative risk reduction in the primary safety endpoint in group F compared with group N (4.7% vs. 6.7%, HR 0.72, 95%CI 0.42-1.65, P = 0.38). The primary efficacy endpoint was 8.0% in group F and 10.0% in group N (HR, 0.82, 95%CI 0.54-1.71, P = 0.49). The composite of the safety and efficacy endpoints at 9 days (10.0% vs. 16.0%, HR 0.61, 95%CI 0.31-1.10, P = 0.10), 30 days (14.0% vs. 17.9%, HR 0.72, 95%CI 0.47-1.16, P = 0.21), or 180 days (18.7% vs. 27.3%, HR 0.65, 95%CI 0.38-1.11, P = 0.11) showed a non-significant trend toward a lower value in group F. CONCLUSION: Fondaparinux resulted in a nonsignificant risk reduction in patients with NSTE-ACS in both bleeding and ischaemic events during short- and long-term follow-up compared with nadroparin.

Thrombus aspiration plus intra-infarct-related artery administration of tirofiban improves myocardial perfusion during primary angioplasty for acute myocardial infarction.

BACKGROUND: We developed a new combined strategy of thrombus aspiration plus intra-infarct-related artery (IRA) bolus administration of tirofiban via the aspiration catheter in patients with ST-segment elevation myocardial infarction (STEMI). This strategy can reduce the distal embolism and achieve highly localized concentrations of tirofiban, which can improve myocardial reperfusion without increasing the risk of bleeding. The aim of this study was to investigate whether this combined strategy is superior to thrombus aspiration alone in improving myocardial perfusion in patients with STEMI undergoing primary angioplasty. METHODS: This single center study included 108 matched control patients with STEMI, angioplasty after thrombus aspiration, and 108 study patients with STEMI plus intra-IRA administration of 500 microg of tirofiban. Both groups had subsequent 12-hour intravenous infusion of 0.1 microg x kg(-1) x min(-1) of tirofiban after angioplasty. The primary end points were Thrombolysis in Myocardial Infarction (TIMI) flow immediately after angioplasty, ST-segment elevation resolution (STR) (> 70%) at 90 minutes after angioplasty, and the peak of creatine kinase-MB (CK-MB) and troponin I (TnI). The secondary end points were the left ventricular ejection fraction (LVEF) in the hospital and at nine months follow-up, cardiac death, target vessel revascularization (TVR), re-infarction and the combination of these three as major adverse cardiac events (MACE) within nine months and any bleeding events. RESULTS: Baseline characteristics of the two groups were well-balanced. The TIMI 3 flow showed a better tendency in the intra-IRA group than in the aspiration alone group (97.22% vs. 87.04%, chi(2) = 7.863, P = 0.049). The peak of CK-MB (83.9 (68.9 - 310.5) U/L vs. 126.1 (74.7 - 356.7) U/L, P = 0.034) and TnI (42.7 (14.7 - 113.9) ng/ml vs. 72.5 (59.8 - 135.3) ng/ml, P = 0.029) were lower in the intra-IRA group than in the aspiration alone group. LVEF in the hospital favored the intra-IRA group, (45.7 +/- 8.3)% to (42.9 +/- 12.1)%, t = 1.98, P = 0.049. There was a tendency towards a lower MACE at 9-month follow-up in the intra-IRA group although it did not reach statistical difference (Log-rank chi(2) = 2.865, P = 0.09). There was no statistical difference in any bleeding events between the two groups. CONCLUSIONS: Thrombus aspiration plus intra-IRA bolus administration of tirofiban combined with angioplasty may be related with improved myocardium perfusion, saved more myocardium, and resulted in a better clinical prognosis.