Molecular recognition of two endogenous hormones by the human parathyroid hormone receptor-1.

Parathyroid hormone (PTH) and PTH-related peptide (PTHrP) are two endogenous hormones recognized by PTH receptor-1 (PTH1R), a member of class B G protein- coupled receptors (GPCRs). Both PTH and PTHrP analogs including teriparatide and abaloparatide are approved drugs for osteoporosis, but they exhibit distinct pharmacology. Here we report two cryo-EM structures of human PTH1R bound to PTH and PTHrP in the G protein-bound state at resolutions of 2.62 Å and 3.25 Å, respectively. Detailed analysis of these structures uncovers both common and unique features for the agonism of PTH and PTHrP. Molecular dynamics (MD) simulation together with site-directed mutagenesis studies reveal the molecular basis of endogenous hormones recognition specificity and selectivity to PTH1R. These results provide a rational template for the clinical use of PTH and PTHrP analogs as an anabolic therapy for osteoporosis and other disorders.

Effects of site-directed mutagenesis of GLP-1 and glucagon receptors on signal transduction activated by dual and triple agonists.

The paradigm of one drug against multiple targets, known as unimolecular polypharmacology, offers the potential to improve efficacy while overcoming some adverse events associated with the treatment. This approach is best exemplified by targeting two or three class B1 G protein-coupled receptors, namely, glucagon-like peptide-1 receptor (GLP-1R), glucagon receptor (GCGR) and glucose-dependent insulinotropic polypeptide receptor for treatment of type 2 diabetes and obesity. Some of the dual and triple agonists have already shown initial successes in clinical trials, although the molecular mechanisms underlying their multiplexed pharmacology remain elusive. In this study we employed structure-based site-directed mutagenesis together with pharmacological assays to compare agonist efficacy across two key signaling pathways, cAMP accumulation and ERK1/2 phosphorylation (pERK1/2). Three dual agonists (peptide 15, MEDI0382 and SAR425899) and one triple agonist (peptide 20) were evaluated at GLP-1R and GCGR, relative to the native peptidic ligands (GLP-1 and glucagon). Our results reveal the existence of residue networks crucial for unimolecular agonist-mediated receptor activation and their distinct signaling patterns, which might be useful to the rational design of biased drug leads.

A high-throughput screening campaign against PFKFB3 identified potential inhibitors with novel scaffolds.

The growth of solid tumors depends on tumor vascularization and the endothelial cells (ECs) that line the lumen of blood vessels. ECs generate a large fraction of ATP through glycolysis, and elevation of their glycolytic activity is associated with angiogenic behavior in solid tumors. 6-Phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) positively regulates glycolysis via fructose-2/6-bisphosphate, the product of its kinase activity. Partial inhibition of glycolysis in tumor ECs by targeting PFKFB3 normalizes the otherwise abnormal tumor vessels, thereby reducing metastasis and improving the outcome of chemotherapy. Although a limited number of tool compounds exist, orally available PFKFB3 inhibitors are unavailable. In this study we conducted a high-throughput screening campaign against the kinase activity of PFKFB3, involving 250,240 chemical compounds. A total of 507 initial hits showing >50% inhibition at 20 µM were identified, 66 of them plus 1 analog from a similarity search consistently displayed low IC50 values (<10 µM). In vitro experiments yielded 22 nontoxic hits that suppressed the tube formation of primary human umbilical vein ECs at 10 µM. Of them, 15 exhibited binding affinity to PFKFB3 in surface plasmon resonance assays, including 3 (WNN0403-E003, WNN1352-H007 and WNN1542-F004) that passed the pan-assay interference compounds screening without warning flags. This study provides potential leads to the development of new PFKFB3 inhibitors.

[Baihe Yuzi Prescription improves asthenospermia by up-regulating the expression of CFTR in the sperm].

OBJECTIVE: To observe the therapeutic efficacy of Baihe Yuzi Prescription (BYP) in the treatment of clinical syndrome-free asthenospermia and its effects on semen parameters, sperm DNA fragmentation index (DFI) and the expression of the cystic fibrosis transmembrane conductance regulator (CFTR) in the sperm. METHODS: We randomly divided 112 patients with clinical syndrome-free asthenospermia into a control group (n = 55) and an experimental group (n = 57), the former treated orally with L-carnitine liquid combined with vitamin E capsules and the latter with BYP in addition, both for 3 months. After treatment, we obtained the total sperm count, sperm motility, percentages of progressively motile sperm (PMS) and morphologically normal sperm (MNS), sperm DFI and expression of CFTR in the sperm, and compared the above parameters between the two groups of patients before and after medication. RESULTS: The total effectiveness rate was significantly higher in the experimental group (82.46%) than in the control (65.45%) (P < 0.05). Compared with the baseline, the patients in the experimental group showed significant improvement after treatment in the total sperm count (��53.5��3.5�� vs ��86.5��3.9�� ��106, P < 0.05), sperm motility (��23.5��3.5��% vs ��38.8��3.7��%, P < 0.05), PMS (��20.1��3.2��% vs ��30.3��3.3��%, P < 0.05), MNS (��2.3��0.3��% vs ��3.9��0.4��%, P < 0.05), sperm DFI (��37.3��3.1��% vs ��25.2��3.4��%, P < 0.05) and the expression of CFTR (P < 0.05), and even better improvement than the controls in sperm motility (��23.8��3.7��% vs ��30.2��3.4��%, P < 0.05), PMS (��19.6��3.1��% vs ��25.3��2.9��%, P < 0.05), MNS (��2.4��0.4��% vs ��3.1��0.3��%, P < 0.05), and sperm DFI (��36.6��3.3��% vs ��30.3��3.1��%, P < 0.05). The total sperm count and the expression of CFTR, however, were not significantly improved in the control group after treatment (P > 0.05). CONCLUSION: Baihe Yuzi Prescription can increase sperm count and motility, improve sperm morphology and DFI in patients with clinical syndrome-free asthenospermia, which may be related to the up-regulated expression of CFTR in the sperm.

GLP-1 mimetics as a potential therapy for nonalcoholic steatohepatitis.

Nonalcoholic steatohepatitis (NASH), as a severe form of nonalcoholic fatty liver disease (NAFLD), is characterized by liver steatosis, inflammation, hepatocellular injury and different degrees of fibrosis. The pathogenesis of NASH is complex and multifactorial, obesity and type 2 diabetes mellitus (T2DM) have been implicated as major risk factors. Glucagon-like peptide-1 receptor (GLP-1R) is one of the most successful drug targets of T2DM and obesity, and its peptidic ligands have been proposed as potential therapeutic agents for NASH. In this article we provide an overview of the pathophysiology and management of NASH, with a special focus on the pharmacological effects and possible mechanisms of GLP-1 mimetics in treating NAFLD/NASH, including dual and triple agonists at GLP-1R, glucose-dependent insulinotropic polypeptide receptor or glucagon receptor.

Signaling profiles in HEK 293T cells co-expressing GLP-1 and GIP receptors.

Glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) are regarded as 'incretins' working closely to regulate glucose homeostasis. Unimolecular dual and triple agonists of GLP-1R and GIPR have shown remarkable clinical benefits in treating type 2 diabetes. However, their pharmacological characterization is usually carried out in a single receptor-expressing system. In the present study we constructed a co-expression system of both GLP-1R and GIPR to study the signaling profiles elicited by mono, dual and triple agonists. We show that when the two receptors were co-expressed in HEK 293T cells with comparable receptor ratio to pancreatic cancer cells, GIP predominately induced cAMP accumulation while GLP-1 was biased towards ß-arrestin 2 recruitment. The presence of GIPR negatively impacted GLP-1R-mediated cAMP and ß-arrestin 2 responses. While sharing some common modulating features, dual agonists (peptide 19 and LY3298176) and a triple agonist displayed differentiated signaling profiles as well as negative impact on the heteromerization that may help interpret their superior clinical efficacies.

Robot-assisted thoracoscopic plication for diaphragmatic eventration.

BACKGROUND: Though conventional thoracoscopic plication is a favorable option of diaphragmatic eventration (DE), ribs limited the movement of trocars, making it difficult to suturing, knot-tying and time-consuming. The purpose of this study was to evaluate delicate surgical maneuvers and suturing time for the management of DE in robot-assisted thoracoscopic plication (RATP). METHODS: From January 2015 to November 2019, 20 patients (14 males; mean age: 10.5 ±â€¯5.2 months; mean weight: 8.6 ±â€¯4.5 kg) who underwent diaphragmatic plication for DE were reviewed at our institution. There were 13 patients with congenital diaphragmatic eventration and 7 patients with acquired diaphragm eventration after congenital heart surgery. RATP was performed on 9 patients (3 on the left and 6 on the right), and conventional thoracoscopic plication (CTP) was applied to 11 patients (5 on the left and 6 on the right). Demographics, the suturing time and complications were respectively evaluated. RESULTS: There was no difference between 2 groups with respect to gender, age at surgery and weight (p > 0.05). No conversion to thoracotomy was needed. The suturing time in RATP group was shorter than CTP group (27.7 ±â€¯3.4 min vs 48.1 ±â€¯4.2 min, p < 0.001). One patient (9.09%) experienced recurrence in CTP group and none was found in RATP group. CONCLUSIONS: Diaphragmatic plication with robot-assisted thoracoscopy or conventional thoracoscopy in DE has minimally invasive and good effect on children. RATP overcome the intercostal limitations to complete delicate suturing and free knot-tying, and has better ergonomics. LEVEL OF EVIDENCE: Level III.

Identification of catalytic and non-catalytic activity inhibitors against PRC2-EZH2 complex through multiple high-throughput screening campaigns.

Enhancer of zeste homolog 2 (EZH2) is the catalytic subunit of the polycomb repressive complex 2 (PRC2) along with embryonic ectoderm development (EED) and suppressor of zeste 12 (SUZ12), which implements transcriptional repression mainly by depositing trimethylation marks at lysine 27 of histone H3 (H3K27me3). Its catalytic activity is closely correlated with the stability of PRC2, and somatic activating mutation of EZH2 Y641F within the catalytic SET domain drives tumor aggressiveness, drug resistance, and poor prognosis. Here, we report two high-throughput screening (HTS) campaigns targeting EZH2 Y641F and EZH2-EED interaction, respectively. For the EZH2 Y641F mutant, the HTS campaign involved a library of 250,000 compounds using a homogenous time-resolved fluorescence (HTRF) assay and identified 162 hits, while 60,160 compounds were screened against EZH2-EED interaction with a fluorescence polarization (FP) assay resulting in 97 hits. Among the 162 EZH2 Y641F inhibitors, 38 also suppressed EZH2-EED interaction and 80 showed inhibitory effects on the wide-type (WT) EZH2. Meanwhile, 10 of the 97 EZH2-EED interaction inhibitors were active against WT EZH2. These hit compounds provide useful tools for the development of novel PRC2-EZH2 inhibitors targeting its catalytic and non-catalytic activities.

An in vitro Förster resonance energy transfer-based high-throughput screening assay identifies inhibitors of SUMOylation E2 Ubc9.

SUMOylation is one of the posttranslational modifications that mediate cellular activities such as transcription, DNA repair, and signal transduction and is involved in the cell cycle. However, only a limited number of small molecule inhibitors have been identified to study its role in cellular processes. Here, we report a Förster resonance energy transfer (FRET) high-throughput screening assay based on the interaction between E2 Ubc9 and E3 PIAS1. Of the 3200 compounds screened, 34 (1.1%) showed higher than 50% inhibition and 4 displayed dose-response inhibitory effects. By combining this method with a label-free surface plasmon resonance (SPR) assay, false positives were excluded leading to discovering WNN0605-F008 and WNN1062-D002 that bound to Ubc9 with KD values of 1.93 ± 0.62 and 5.24 ± 3.73 µM, respectively. We examined the effect of the two compounds on SUMO2-mediated SUMOylation of RanGAP1, only WNN0605-F008 significantly inhibited RanGAP1 SUMOylation, whereas WNN1062-D002 did not show any inhibition. These compounds, with novel chemical scaffolds, may serve as the initial material for developing new SUMOylation inhibitors.

Transumbilical enterostomy for Hirschsprung's disease with a two-stage laparoscopy-assisted pull-through procedure.

BACKGROUND: A one-stage laparoscopic operation has recently been considered a favorable option for the management of patients with Hirschsprung's disease (HD) due to its superior cosmetic results. One-stage transanal endorectal pull-through for the treatment of rectosigmoid HD has been widely used in newborns without complications. However, enterostomy is required in some HD cases for enterocolitis and dilated colon. Our transumbilical enterostomy (TUE) and two-stage laparoscopy-assisted anorectoplasty were effective and achieved a similar cosmetic effect to one-stage laparoscopy on the abdominal wall in patients with anorectal malformation, but the effect in patients with HD is unclear. AIM: To evaluate the safety, efficacy and cosmetic results of TUE in two-stage laparoscopy-assisted pull-through for HD. METHODS: From June 2013 to June 2018, 53 patients (40 boys, 13 girls; mean age at enterostomy: 5.5 ± 2.2 mo) who underwent enterostomy and two-stage laparoscopy-assisted pull-through for HD with stoma closure were reviewed at our institution. Two enterostomy approaches were used: TUE in 24 patients, and conventional abdominal enterostomy (CAE) in 29 patients. Eleven patients with rectosigmoid HD had severe preoperative enterocolitis or a dilated colon. 26 patients had long-segment HD, and 16 patients had total colonic aganglionosis (TCA). The patients with left-sided HD underwent the two-stage laparoscopic Soave procedure, and the patients with right-sided HD and TCA underwent the laparoscopic Duhamel procedure. Demographics, enterostomy operative time, complications and cosmetic results were respectively evaluated. RESULTS: There were no differences between the groups with respect to gender, age at enterostomy, weight and clinical type (P > 0.05). No conversion to open technique was required. Two patients experienced episodes of stomal mucosal prolapse in the TUE group and 1 patient in the CAE group (8.33% vs 3.45%, P > 0.05). No parastomal hernia was observed in either of the two groups. Wound infection at the stoma was seen in 1 case in the TUE group, and 2 cases in the CAE group (4.17% vs 6.90%, P > 0.05). No obstruction was noted in any of the patients in the TUE group, whereas obstruction was found in 1 patient in the CAE group. Enterocolitis was observed in 3 and 5 patients in the TUE and CAE group, respectively (12.50% vs 17.24%, P > 0.05). There was no significant difference between the TUE group and CAE group in terms of the incidence of soiling and constipation (P > 0.05). The cosmetic result using the scar score in the TUE group was better than that in the CAE group (6.83 ± 0.96 vs 13.32 ± 1.57, P < 0.05). CONCLUSION: TUE is a safe and feasible method for the treatment of HD, and the staged enterostomy and two-stage laparoscopy-assisted pull-through achieved a similar cosmetic effect to the one-stage laparoscopic procedure.

Identification, structure modification, and characterization of potential small-molecule SGK3 inhibitors with novel scaffolds.

The serum and glucocorticoid-regulated kinase (SGK) family has been implicated in the regulation of many cellular processes downstream of the PI3K pathway. It plays a crucial role in PI3K-mediated tumorigenesis, making it a potential therapeutic target for cancer. SGK family consists of three isoforms (SGK1, SGK2, and SGK3), which have high sequence homology in the kinase domain and similar substrate specificity with the AKT family. In order to identify novel compounds capable of inhibiting SGK3 activity, a high-throughput screening campaign against 50,400 small molecules was conducted using a fluorescence-based kinase assay that has a Z' factor above 0.5. It identified 15 hits (including nitrogen-containing aromatic, flavone, hydrazone, and naphthalene derivatives) with IC50 values in the low micromolar to sub-micromolar range. Four compounds with a similar scaffold (i.e., a hydrazone core) were selected for structural modification and 18 derivatives were synthesized. Molecular modeling was then used to investigate the structure-activity relationship (SAR) and potential protein-ligand interactions. As a result, a series of SGK inhibitors that are active against both SGK1 and SGK3 were developed and important functional groups that control their inhibitory activity identified.

High-throughput screening campaigns against a PI3Kα isoform bearing the H1047R mutation identified potential inhibitors with novel scaffolds.

The phosphatidylinositol 3-kinase (PI3K) pathway is involved in many cellular functions including cell growth, metabolism, and transformation. Hyperactivation of this pathway contributes to tumorigenesis, therefore, PI3K is a major target for anticancer drug discovery. Since the PI3Kα isoform is implicated mostly in cancer, we conducted a high-throughput screening (HTS) campaign using a 3-step PI3K homogenous time-resolved fluorescence assay against this isoform bearing the H1047R mutation. A total of 288,000 synthetic and natural product-derived compounds were screened and of which, we identified 124 initial hits that were further selected by considering the predicted binding mode, relationship to known pan-assay interference compounds and previous descriptions as a lipid kinase inhibitor. A total of 24 compounds were then tested for concentration-dependent responses. These hit compounds provide novel scaffolds that can potentially be optimized to create novel PI3K inhibitors.

Laparoscopic-assisted Duhamel procedure with ex-anal rectal transection for total colonic aganglionosis.

PURPOSE: Laparoscopic-assisted Duhamel procedure has a larger anastomosis and a reservoir which allows early recovery of defecation frequency, but concerns have been raised regarding the long operative time, high incidence of pouchitis and Hirschsprung associated enterocolitis (HAEC). The purpose of this study was to evaluate the postoperative complications and functional outcomes for patients with TCA undergoing modified laparoscopic-assisted Duhamel procedure (MLDP) with ex-abdominal partial colectomy and ex-anal rectal transection. METHODS: From 2011 to 2014, 16 patients with TCA who underwent MLDP were reviewed at our institution. Main modified techniques were to mobilize partial bowel through abdominal stoma opening, mobilize remaining colon, and dissect the retro-rectal space using laparoscopy, pull out and transect rectum ex-anally using a linear stapling device for creation of a short rectal pouch of 35~45mm. Seven patients who underwent classical laparoscopic Duhamel procedure (CLDP) with a long rectal pouch of 50-60mm between 2009 and 2011 were used as control group. Data were collected including demographics, laparoscopic technique, operative time, stool frequency, complications and continence outcomes. RESULTS: The operative time in MLDP group was significantly shorter than control group (3.0h vs. 4.7h, p=0.02). The incidence of postoperative HAEC in MLDP group was lower than control group (12.5% versus 42.9%; p=0.03) within the second postoperative year. Two patients (28.6%) experienced episodes of pouchitis in CLDP group and none was found in MLDP group. There was no significant difference in overall functional outcome between two groups, but the performance of MLDP group was better in terms of diapers required than CLDP group (1.80±0.45 vs. 1.00±0.64; p=0.02). All patients after 4years of age had a normal defecation frequency in both groups. CONCLUSIONS: MLDP is a safe, simple, and reliable technique for TCA. It has fewer postoperative complications due to the short rectal pouch. However, longer follow-up and a larger sample size are necessary to prove the efficacy in the treatment of TCA. LEVEL OF EVIDENCE: Level 3.

Laparoscopic finding of a hepatic subcapsular spider-like telangiectasis sign in biliary atresia.

AIM: To assess the diagnostic value of a laparoscopic finding of a hepatic subcapsular spider-like telangiectasis (HSST) sign in biliary atresia. METHODS: A retrospective study was conducted first and then a validation set was used to investigate the value of an HSST sign in predicting biliary atresia (BA). In the retrospective study, laparoscopic images of the liver surface were reviewed in 126 patients with infantile cholestasis (72 BA patients and 54 non-BA cholestasis patients) and a control group of 38 patients with non-hepatic conditions. Analysis was first made by two observers separately and finally, a consensus conclusion was achieved. Then, the diagnostic value of the HSST sign was validated in an independent cohort including 45 BA and 45 non-BA patients. RESULTS: In the retrospective investigation, an amplified HSST sign was found in all BA patients, while we were unable to detect the HSST sign in 98.1% of the 54 non-BA patients. There was no HSST sign in any of the control subjects. In the first review, the sensitivity and specificity from one reviewer were 100% and 98.1%, respectively, and the results from the other reviewer were both 100%. The consensus sensitivity and specificity were 100% and 98.1%, respectively. The HSST sign was defined as being composed of several enlarged tortuous spider-like vascular plexuses with two to eight branches distributed on all over the liver surface, which presented as either a concentrated type or a dispersed type. In the independent validation group, the sensitivity, specificity, positive predictive value and negative predictive value of the HSST sign were 100%, 97.8%, 97.8% and 100%, respectively. CONCLUSION: The HSST sign is characteristic in BA, and laparoscopic exploration for the HSST sign is valuable in the diagnosis of BA.

Laparoscopic Duhamel Procedure with Ex-Anal Rectal Transection for Right-Sided Hirschsprung's Disease.

BACKGROUND: Increased defecation frequency and soiling are common complications of surgery for right-sided Hirschsprung's disease (HD). Though the laparoscopic Duhamel procedure is a favorable option in right-sided HD, the conventional laparoscopic technique is time consuming and has complications that are associated with the reservoir. In this study, we described a modified laparoscopic Duhamel technique with ex-anal rectal transection combined with the Deloyer's procedure for right-sided HD. MATERIALS AND METHODS: Between March 2010 and January 2015, 18 right-sided HD underwent this modified laparoscopic Duhamel procedure (MLDP). The main modifications were to mobilize the colon and to dissect the retrorectal space under laparoscopy, pull out, and transect ex-anally by using a linear stapling device through a transverse incision on posterior wall of the rectum. Seven age-matched patients who underwent a conventional laparoscopic Duhamel procedure (CLDP) between March 2008 and December 2010 were included as the control group. Demographics, surgical procedures, operative data, postoperative complications, and clinical outcomes were compared between these two groups. RESULTS: Operative time was significantly shorter in the MLDP group (150 minutes versus 195 minutes; P = .03). There was no significant difference in mean timespan to obtain a normal defecation frequency (3.8 months versus 3.6 months), incidence of soiling (11.1% versus 14.3%; P = .34), and constipation (5.6% versus 14.3%; P = .69). Pouchitis were not found in the MLDP group, whereas 2 patients in the CLDP group (28.6%) presented with pouchitis and intractable diarrhea, requiring spur division. The incidence of postoperative enterocolitis was not significantly different in the two groups (11.1% in MLDP versus 14.3% in CLDP; P = .68). All 22 patients had a normal defecation over 4 years of age. CONCLUSIONS: Laparoscopic Duhamel with ex-anal rectal transection is a simple, easy-to-learn, and effective procedure for right-sided HD. It has low postoperative pouchitis because of a short pouch.

Association of Ureaplasma urealyticum colonization with development of bronchopulmonary dysplasia: a systemic review and meta-analysis.

There is controversy regarding the roles of Ureaplasma urealyticum (U. urealyticum) colonization in the development of bronchopulmonary dysplasia (BPD). This study explored the association between U. urealyticum and bronchopulmonary dysplasia at 36 weeks post-menstrual age (BPD36). Studies published before December 31, 2013 were searched from Medline, Embase, Ovid, Web of Science, and Cochrane databases, with the terms "Ureaplasma urealyticum", "chronic lung disease", or "BPD36" used, and English language as a limit. The association between U. urealyticum colonization and BPD36 was analyzed with RevMan 4.2.10 software, using the odds ratio (OR) and relative risk (RR) for dichotomous variables. Out of the enrolled 81 studies, 11 investigated the BPD36 in total 1193 infants. Pooled studies showed no association between U. urealyticum colonization and subsequent development of BPD36, with the OR and RR being 1.03 (95% CI=0.78-1.37; P=0.84) and 1.01 (95% CI= 0.88-1.16, P=0.84), respectively. These findings indicated no association between U. urealyticum colonization and the development of BPD36.

[The study of different operative style for abdominal aortic aneurysm during surrounding operation].

OBJECTIVE: To compare the the similarities and differences during the surrounding operation of endovascular repair (EVAR) and open surgical repair (OSR) for abdominal aortic aneurysm. METHODS: 112 patients with abdominal aortic aneurysms (AAA) were selected from 2004 to 2009: among them, 66 patients were treated with EVAR, 46 patients with OSR. Data of two groups were collected and analyzed during surrounding operation. RESULTS: Compared to OSR group, the mean blood lost, blood transfusion and intra-operative fluid in EVAR group were significantly less than OSR group (P < 0.05). The mean time of operation, observation period in ICU and being in hospital in EVAR group were shorter than OSR group (P < 0.05). But the cost of hospitalization in EVAR was far higher than that of OSR group (P < 0.05). In short term postoperative complications the OSR group was higher than the EVAR (P < 0.05), however, there was no statistically significant difference in death rate of the two groups during surrounding operation (P > 0.05). CONCLUSION: EVAR has the advantages of mild trauma, less blood loss, quicker recovery after operation, and less disturbance to internal environment. Especially, it is suitable for the patients who can not undergo open surgery repair, but its cost is still higher.

[Analysis of death-related factors of type B aortic dissection treated medically during the acute phase].

OBJECTIVE: To analyze the death-related risk factors of type B aortic dissection treated medically during the acute phase (symptoms presenting within 14 d), and to determine the predictors of surgical indications for acute type B aortic dissection. METHODS: Clinical data of 42 patients with acute type B aortic dissection admitted from January 2007 to May 2009 was retrospectively reviewed. There were 33 male and 9 female with a mean age of (50 +/- 12) years old. Therapy included analgesia, controlled hypotension and beta-receptor blocker, the mortality in acute phase was 33.3% (14/42). Univariate and multivariate logistic regression analyses were performed to identify the predictors of the death in acute phase. RESULTS: In univariate logistic regression analysis, the malperfusion of aortic branches (P = 0.018) and maximum aortic diameter (P = 0.002) were significant predictors of death. In the multivariate logistic regression model, the malperfusion of aortic branches (P = 0.041) and maximum aortic diameter (P = 0.005) were also considered as the significant death-related factors.Risk of death augmented significantly (P = 0.000) when the maximum aortic diameter over 40 mm. CONCLUSION: Malperfusion of aortic branches and the large maximum aortic diameter (> 40 mm) are the indications of surgery or endovascular therapy for acute type B aortic dissection.

[Expression of proto-oncogene Wip1 in breast cancer and its clinical significance].

OBJECTIVE: To investigate the expression of proto-oncogene Wip1 in breast cancer tissue and its clinical significance. METHODS: Through the uses of semi-RT-PCR, immunohistochemical technique and Western blot, the specimens from 70 patients of breast cancer and 20 normal controls were detected for Wip1 mRNA and protein expression. At the same time, the authors analyzed the relations between the expression of Wip1 in human breast cancer and different clinical pathologic parameters. RESULTS: RT-PCR: The values of gene expression of Wip1 mRNA in breast cancer tissue, pericancerous tissue and normal breast tissue were 0.715 +/- 0.087, 0.175 +/- 0.021 and 0.154 +/- 0.022 respectively. Thus the value of gene expression in breast cancer tissue was significantly higher than that in pericancerous tissue or normal breast tissue (P < 0.01). Immunohistochemistry: The high expression rates of Wip1 protein in breast cancer tissue, pericancerous tissue and normal breast tissue were 62.9% (44/70), 2.9% (2/70) and 0 (0/20) respectively and there was a significant difference among these three different tissues (P < 0.01). Western blot: The relative contents of Wip1 protein in breast cancer tissue, pericancerous tissue and normal breast tissue were 0.688 +/- 0.151, 0.251 +/- 0.043 and 0.234 +/- 0.044 respectively. The relative content of Wip1 protein in breast cancer tissue was significantly higher than that in pericancerous tissue or normal breast tissue (P < 0.01). The high expression of Wip1 protein was negatively correlated with the expression of p53, but it had nothing to do with tumor size, age, tumor staging, axillary lymph node metastasis and expressions of ER and PR. CONCLUSION: The high expression of Wip1 mRNA and its protein in breast cancer tissue may promote the growth of breast cancer. Wip1 may become a new target for therapy of breast cancer.