Effect of quercetin on granulosa cells development from hierarchical follicles in chicken.

1. The bioflavonoid quercetin is a biologically active component, but its functional regulation of granulosa cells (GCs) during chicken follicular development is little studied. To investigate the effect of quercetin on follicular development in laying hens, an in vitro study was conducted on granulosa cells from hierarchical follicles treated with quercetin.2. The effect of quercetin on cell activity, proliferation and apoptosis of granulosa cells was detected by CCK-8, EdU and apoptosis assays. The effect on progesterone secretion from granulosa cells was investigated by enzyme-linked immunosorbent assay (ELISA). Expression of proliferating cell nuclear antigen (PCNA) mRNA and oestrogen receptors (ERs), as well as the expression of steroid acute regulatory protein (StAR), cytochrome P450 cholesterol side chain cleavage enzyme (P450scc) and 3ß-hydroxysteroid dehydrogenase (3ß-HSD) mRNA during progesterone synthesis, were measured by real-time quantitative polymerase chain reaction (RT-qPCR). PCNA, StAR and CYP11A1 protein expression levels were detected using Western blotting (WB).3. The results showed that treatment with quercetin in granulosa cells significantly enhanced cell vitality and proliferation, reduced apoptosis and promoted the expression of gene and protein levels of PCNA. The levels of progesterone secretion increased significantly following quercetin treatment, as did the expression levels of StAR and CYP11A1 using the Western Blot (WB) method.4. The mRNA expression levels of ERα were significantly upregulated in the 100 ng/ml and 1000 ng/ml quercetin-treated groups, while there was no significant difference in expression levels of ERß mRNA.

[The effect of Wendler Glottoplasty to elevate vocal pitch in transgender women].

Objective: To evaluate the effect of Wendler Glottoplasty to elevate vocal pitch in transgender women. Methods: The voice parameters of pre-and 3-month post-surgery of 29 transgender women who underwent Wendler Glottoplasty in department of otorhinolaryngology head and neck surgery of Beijing Friendship Hospital from January, 2017 to October, 2020 were retrospectively analyzed. The 29 transgender women ranged in age from 19-47 (27.0±6.3) years old. Subjective evaluation was performed using Transsexual Voice Questionnaire for Male to Female (TVQMtF). Objective parameters included fundamental frequency (F0), highest pitch, lowest pitch, habitual volume, Jitter, Shimmer, maximal phonation time (MPT), noise to harmonic ratio (NHR) and formants frequencies(F1, F2, F3, F4). SPSS 25.0 software was used for statistically analysis. Results: Three months after surgery, the score of TVQMtF was significantly decreased [(89.9±14.7) vs. (50.4±13.6), t=11.49, P<0.001]. The F0 was significantly elevated [(152.7±23.3) Hz vs. (207.7±45.9) Hz, t=-6.03, P<0.001]. Frequencies of F1, F2 and F3 were significantly elevated. No statistical difference was observed in the frequencies of F4. The highest pitch was not significantly altered while the lowest pitch was significantly elevated [(96.8±17.7) Hz vs. (120.0±28.9) Hz, t=-3.71, P=0.001]. Habitual speech volume was significantly increased [(60.0±5.2) dB vs. (63.6±9.6) dB, t=-2.12, P=0.043]. Jitter, Shimmer, NHR and MPT were not obviously altered (P>0.05). Conclusions: Wendler Glottoplasty could notably elevate the vocal pitch, formants frequencies and degree of vocal femininity in transgender women without affecting phonation ability and voice quality. It can be an effective treatment modality for voice feminization.

[Construction of a nomogram model for predicting 2-year survival rate of small cell lung cancer based on more comprehensive variables].

Objective: To construct a novel prognostic nomogram model based on more comprehensive variables for patients with small-cell lung cancer (SCLC). Methods: The data of 722 patients with SCLC confirmed by pathology in Affiliated Cancer Hospital of Shanxi Medical University from January 2015 to December 2018 were retrospectively analyzed [including 592 males and 130 females, aged from 23 to 82(61±9) years]. A random seed count of 133 was used to divide those patients into training set (n=422) and validation set (n=300). Kaplan-Meier was used for survival curves analysis and univariate Log-rank test was used for evaluating the influence of clinical variables on the prognosis of sclc, variables with P<0.05 in univariate analysis were included in a multivariate Cox regression model. The nomogram was constructed based on the variables which P<0.05 in multivariate analysis. Receiver operating characteristic (ROC) curve, calibration by Integrated Brier score (IBS) and clinical net benefit by decision curve analysis (DCA) were used to evaluate model discriminative power, prediction error value, and clinical net benefit, and compared with the American Joint Committee on Cancer 8th TNM. Results: Male, abnormal monocyte (MON) counts, abnormal neuron specific enolase (NSE), abnormal cytokeratin 19 fragment (Cyfra211), M1a stage, M1b stage, M1c stage, radiotherapy (RT), chemotherapy ≥4 cycles and prophylactic cranial irradiation (PCI) were prognostic factors for SCLC[HR(95%CI)=1.39(1.00-1.92), 1.29(1.02-1.63), 1.41(1.11-1.80), 2.02(1.48-2.76), 1.09(0.77-1.55), 1.44(0.94-2.22), 2.01(1.49-2.71), 0.75(0.57-0.98), 0.40(0.31-0.51)and 0.42(0.26-0.68), respectively, all P<0.05]. The area under ROC curve (AUC) of the nomogram in training set and validation set were 0.814(95%CI: 0.765-0.862)and 0.787 (95%CI: 0.725-0.849), which were higher than TNM [0.616(95%CI: 0.558-0.674) and 0.648(95%CI: 0.581-0.715)].The calibration curve showed a good correlation between the nomogram prediction and actual observation for the 2-year overall survival (OS). IBS indicted a lower prediction error rate (training set: 0.132 vs 0.169; validation set: 0.138 vs 0.169). DCA showed a wider threshold range than TNM (training set: 0.01-0.96 vs 0.01-0.85, validation set: 0.01-0.94 vs 0.01-0.86) and a greater improvement of the clinical net benefit (in training set the nomogram had a greater clinical benefit than TNM in the range of 0.19-0.96, and remained in validation set in the range of 0.19-0.94). Conclusion: The established nomogram model for predicting 2-year OS in patients with SCLC based on 8 variables, including gender, MON, NSE, Cyfra211, M stage, RT, CT cycles and PCI can be used for an more accurately prognosis prediction and reference for therapeutic regimen selection.

Short-term lactation and mammary metabolism responses in lactating goats to graded removal of methionine from an intravenously infused complete amino acid mixture.

To investigate the possible pathways of Met deficiency to depress milk protein synthesis, 4 lactating goats fitted with jugular vein, mammary vein, and carotid artery catheters and transonic blood flow detectors on the external pudic artery were used in a 4 × 4 Latin square experiment. Goats were fasted for 24 h followed by a 9-h intravenous infusion of an AA mixture plus glucose. Milk yield was recorded and samples were taken in h 2 to 8 of the infusion period, and mammary biopsy was performed in the last hour. Treatments were graded removal of Met from the infused AA mixture to achieve Met content in the infusate of 100 (complete), 60, 30, or 0% of that in casein. Graded Met removal decreased yield of milk, milk protein, and lactose linearly and tended to decrease yield of milk fat linearly. Milk protein yield decreased to 82, 78, and 69% that of complete mixture infusion, respectively, when the 60, 30, and 0% Met infusate was infused. Circulating Met decreased linearly with graded Met removal. Arterial and venous Met decreased to 36 and 23% that of complete mixture infusion, respectively, when all Met was removed out of the mixture. Concomitant with the decreased circulating concentration was a similar increase in mammary Met affinity as reflected by the linearly increased mammary Met clearance rate. The increased affinity plus the linearly increased mammary blood flow totally offset the negative effect of decreased circulating Met concentration on mammary Met uptake. The overall result was similar mammary Met uptakes across treatments ranging from 285.9 to 339.5 µmol/h. Mammary uptakes of the other AA measured were generally not affected by treatments except for a linearly decreased Thr uptake and a trend of linearly increased Glu uptake. Consistent with the behavior of an AA mainly catabolized in the liver and mainly used for protein synthesis in peripheral tissues, mammary uptake to milk output ratios of Met measured in the present study ranged from 1.25 to 1.49 and was not affected by treatments. For the other AA measured, the ratio of Thr was linearly decreased and that of Glu was linearly increased by graded Met removal. Graded Met removal linearly elevated circulating urea N and glucose concentrations, indicating enhanced whole-body catabolism of AA and hepatic gluconeogenesis. Treatments had no significant effects on circulating insulin, growth hormone, and the other hormones and metabolites measured. Phosphorylation status of eIF4E binding protein 1 tended to decrease linearly and that of p70S6k was linearly decreased by graded Met removal, indicating depressed signal in the intracellular mechanistic target of rapamycin complex 1 (mTORC1) signaling pathway. In conclusion, results of the present study indicated that the mTORC1 pathway and whole-body AA catabolism rather than mammary uptake appeared the drivers for changes in milk protein synthesis in response to varying Met supply.

[Clinical analysis of 234 esophageal foreign bodies].

Objective: To explore the clinical characteristics and treatment methods of esophageal foreign body. Method: The clinical data of 234 patients with esophageal foreign bodies admitted to our department from January 2015 to August 2018 were retrospectively analyzed, including course time, foreign body types, surgical methods, imaging manifestations and treatment related complications. Result: The diagnosis of esophageal foreign bodies was confirmed by esophageal CT or esophageal barium meal X-ray examination before operation in 234 patients. Course time varied from 3 hours to 7 days, and the jujube nucleus was the most common food-borne foreign body.223 patients underwent esophagoscopic exploration and foreign body removal under general intravenous anesthesia, 11 of them had no definite esophageal foreign body, 22 had esophageal perforation and periesophagitis. After removal of foreign body, the nasogastric feeding tube was inserted. About 10 days later, the nasogastric feeding tube was removed when they got healthy. Nine cases underwent cervical abscess incision and drainage under general anesthesia. The average postoperative hospital day was 11 days. Conclusion: The rigid esophagoscopy is a safe and effective method for the esophageal foreign bodies. And neck abscess incision must be necessary,when they suffered from esophageal perforation with neck abscess and other serious complications.

CalliSpheres® drug-eluting beads (DEB) transarterial chemoembolization (TACE) is equally efficient and safe in liver cancer patients with different times of previous conventional TACE treatments: a result from CTILC study.

PURPOSE: To assess the efficacy and safety of drug-eluting beads transarterial chemoembolization (DEB-TACE) in liver cancer patients with different times of previous conventional transarterial chemoembolization (cTACE) treatments. METHODS: 367 liver cancer patients about to receive DEB-TACE treatment were enrolled in this prospective cohort study. All patients were divided into no previous cTACE group (NPC group), 1-2 times previous cTACE group (PC group) and triple or above previous cTACE group (TPC group) according to the times of previous cTACE treatments. RESULTS: There was no difference in complete response (CR) (P = 0.671) and objective response rate (ORR) (P = 0.062) among three groups. Additionally, no difference in overall survival (OS) among groups (P = 0.899) was found. As to liver function, most liver function indexes were deteriorative at 1 week after DEB-TACE operation, but returned to baseline at 1-3 months after DEB-TACE operation in all three groups, while percentage of abnormal total bile acid (TBA) patients was higher in TPC group than NPC and PC groups at 1-3 month post-DEB-TACE (P = 0.018). As for safety profiles, the incidence of pain during DEB-TACE operation was lower in TPC group compared to NPC and PC groups (P = 0.005), while no difference of other adverse events was found during and 1 month post-DEB-TACE treatment among three groups. CONCLUSION: DEB-TACE treatment was equally efficient and tolerated in liver cancer patients with different times of previous cTACE treatments.

Subtype-specific inherited predisposition to pemphigus in the Chinese population.

BACKGROUND: Pemphigus is a group of rare life-threatening mucocutaneous autoimmune diseases, presenting mainly as two subtypes: pemphigus vulgaris (PV) and pemphigus foliaceus (PF). Inherited predispositions to pemphigus have long been speculated but they remain poorly understood. OBJECTIVES: To identify common and specific nongenetic and genetic factors associated with pemphigus and its subtypes in the Chinese population. METHODS: A genome-wide association study (GWAS) was performed in 496 unrelated patients with pemphigus (including 365 with PV and 104 with PF) and 1105 controls without pemphigus. RESULTS: A sex preference was observed only in PV (57·5% female) and not in PF (47·1% female). For male patients only, the mean age at diagnosis was significantly lower for PV than for PF (P < 0·001). The strongest associated single-nucleotide polymorphisms are in the human leucocyte antigen (HLA) region: rs70993900 (PV; P = 1·5 × 10-45 ) and rs9469220 (PF; P = 1·1 × 10-8 ). HLA-DQB1*05:03 ranks at the top (P = 4·7 × 10-40 ; odds ratio 12·4) in both subtypes, with significantly different risk allele frequency (RAFPV = 34·2% vs. RAFPF = 18·8% vs. RAFcontrol = 4·4%), whereas HLA-DRB1*14:01 and HLA-DRB1*04:06 are PV specific. HLA-DQB1*03:03 and HLA-DQB1*03:02 show significant subtype specificity in opposite directions. All of these associations were validated in the replication series with 147 cases of pemphigus and 604 controls. Multiple novel non-HLA susceptibility loci were also identified in the GWAS. CONCLUSIONS: This study represents the largest GWAS on pemphigus in the Chinese population published to date, and has allowed us to identify HLA haplotypes significantly shared between or specific to the two main subtypes of pemphigus.

Removal of epicardial adipose tissue after myocardial infarction improves cardiac function.

BACKGROUND: The aim of this study was to assess the role of lipolysis of epicardial adipose tissue (EAT) in cardiac function after myocardial infarction (MI). METHODS: We used a rat model of MI with or without EAT removal to study the effects of EAT lipolysis on cardiovascular function. Echocardiography and cardiac catheterization were used to determine cardiac function, and infarct size and histopathology specimens were analyzed in postmortem sections. Inflammatory responses were evaluated via flow cytometry and Elisa analyses. RESULTS: We found that the lipolysis of EAT increased significantly after MI. Removal of the EAT after MI (MI-EAT) improved cardiac function by nearly 10% and decreased the infarct area by 6% when compared with rats retaining EAT after MI (MI+EAT). Furthermore, the removal of EAT reduced the number of CD45-positive leukocytes (50 vs. 34.8%) and increased the ratio of macrophage/leukocytes (56 vs. 75%) in the infarcted heart. Compared with the MI+EAT group, the concentration of tumor necrosis factor-alpha and interleukin 1­beta were reduced in the MI-EAT group. CONCLUSION: Lipolysis of EAT increased significantly after MI. Removal of EAT improved cardiac function, in part, by weakening the inflammatory response.

[Effect of Δ40p53 isoform on enhancing the pro-apoptotic function of p53 in tumor cells].

Objective: To investigate the effect of Δ40p53, an alternative spliced isoform of p53 lacking the N-ter minus, on the pro-apoptotic function of p53. Methods: The wild-type p53 was ectopically expressed in HCT116-p53(-/-) (endogenous Δ40p53 expression), HCT116-p53(+ /+) (wild-type p53) and H1299 (p53-null) cells by adenoviral delivery, while Δ40p53 plasmid were transfected into these cells to overexpress Δ40p53. The levels of Δ40p53 and p53 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR) and quantitative PCR. The expression of related proteins was deter mined by Western blotting. The interaction of p53 and Δ40p53 was observed by co-immunoprecipitation assay. Calcein-AM/propidium iodide (PI) staining and flow cytometry were used to detect the apoptotic rate of tested cells in each group. Results: HCT116-p53(-/-) cells expressed endogenous Δ40p53 isoform. Neither transcription nor protein expression of wild-type p53 was interfered by the increased expression of Δ40p53. Full length p53 and Δ40p53 could bind to each other. Calcein-AM/PI staining showed that the apoptotic rates of H1299-Control, HCT116-p53(-/-) -Control, H1299+ p53, HCT116-p53(-/-)+ p53, H1299+ oxaliplatin (Oxa), HCT116-p53(-/-)+ Oxa, H1299+ p53+ Oxa and HCT116-p53(-/-)+ p53+ Oxa groups were (2.50±0.47)%, (2.40±0.32)%, (5.20±0.58)%, (4.10±0.18)%, (22.40±1.73)%, (19.30±1.11)%, (29.90±1.15)% and (39.30±2.26)%, respectively. It was statistically significant between H1299+ p53+ Oxa and HCT116-p53(-/-)+ p53+ Oxa groups (t=3.721, P=0.0205). Moreover, the apoptotic rates of H1299-Control, H1299+ Δ40p53, H1299+ p53, H1299+ p53+ Δ40p53, H1299+ Oxa, H1299+ Δ40p53+ Oxa, H1299+ p53+ Oxa and H1299+ p53+ Δ40p53+ Oxa groups were (2.60±0.35)%, (2.20±0.17)%, (4.80±0.49)%, (4.90±1.10)%, (20.30±1.10)%, (19.60±1.45)%, (27.90±1.39)%, (35.20±1.43)%, respectively. Furthermore, flow cytometry assay showed that the apoptotic rates of above cells were (2.70±0.32)%, (2.20±0.24)%, (4.60±0.48)%, (3.90±0.67)%, (19.30±1.11)%, (17.70±0.66)%, (28.30±2.76)% and (37.50±1.51)%, respectively. H1299+ p53+ Δ40p53+ Oxa cells showed higher cell apoptosis than H1299+ p53+ Oxa cells (t=2.930, P=0.042). Conclusion: Δ40p53 isoform can bind to full-length p53, and enhance its pro-apoptotic function in tumor cells.

Effects of graded removal of lysine from an intravenously infused amino acid mixture on lactation performance and mammary amino acid metabolism in lactating goats.

To investigate responses of milk protein synthesis and mammary AA metabolism to a graded decrease of postruminal Lys supply, 4 lactating goats fitted with jugular vein, mammary vein, and carotid artery catheters and transonic blood flow detectors on the external pudic artery were used in a 4 × 4 Latin square experiment. Goats were fasted for 24 h and then received a 9-h intravenous infusion of an AA mixture plus glucose. Milk yield was recorded and samples were taken in h 2 to 8 of the infusion period; a mammary biopsy was performed in the last hour. Treatments were graded decrease of lysine content in the infusate to 100 (complete), 60, 30, or 0% as in casein. Lysine-removed infusions linearly decreased milk yield, tended to decrease lactose yield, and tended to increase milk fat to protein ratio. Milk protein content and yield were linearly decreased by graded Lys deficiency. Mammary Lys uptake was concomitantly decreased, but linear regression analysis found no significant relationship between mammary Lys uptake and milk protein yield. Treatments had no effects on phosphorylation levels of the downstream proteins measured in the mammalian target or rapamycin pathway except for a tended quadratic effect on that of eukaryotic initiation factor 2, which was increased and then decreased by graded Lys deficiency. Removal of Lys from the infusate linearly increased circulating glucagon and glucose. Removal of Lys from the infusate linearly decreased arterial and venous concentrations of Lys. Treatments also had a significant quadratic effect on venous Lys, suggesting mechanisms to stabilize circulating Lys at a certain range. The 2 infusions partially removing Lys resulted in a similar 20% decrease, whereas the 0% Lys infusion resulted in an abrupt 70% decrease in mammary Lys uptake compared with that of the full-AA mixture infusion. Consistent with the abrupt decrease, mammary Lys uptake-to-output ratio decreased from 2.2 to 0.92, suggesting catabolism of Lys in the mammary gland could be completely prevented when the animal faced severe Lys deficiency. Mammary blood flow was linearly increased, consistent with the linearly increased circulating nitric oxide by graded Lys deficiency, indicating mechanisms to ensure the priority of the mammary gland in acquiring AA for milk protein synthesis. Infusions with Lys removed increased mammary clearance rate of Lys numerically by 2 to 3 fold. In conclusion, the decreased milk protein yield by graded Lys deficiency was mainly a result of the varied physiological status, as indicated by the elevated circulating glucagon and glucose, rather than a result of the decreased mammary Lys uptake or depressed signals in the mTOR pathway. Mechanisms of Lys deficiency to promote glucagon secretion and mammary blood flow and glucagon to depress milk protein synthesis need to be clarified by future studies.

[The research progress of LGR4 in the eye development and related diseases].

LGR4, also known as GPR48, is a member of the leucine-rich, G protein-coupled receptor family. It is widely expressed in tissues of the reproductive system, urinary system, sensory organs, digestive system, and central nervous system. LGR4 plays an important role in the development of various organs and cancer development and progression by modulating multiple signaling pathways. Recent studies have revealed that LGR4 is related with many kinds of human diseases such as gastrointestinal carcinomas. Eye development is a dynamic process regulated by a number of growth factors and cytokines. LGR4 is extensively expressed in the eyes in a finely tuned spatiotemporal pattern. Mice lacking LGR4 have been found to display anterior segment dysgenesis, including microphthalmia, iris hypoplasia, iridocorneal angle malformation and corneal dysgenesis, cataract and other defects. Here we review the role of LGR4 in the eye development and related molecular mechanisms. (Chin J Ophthalmol, 2017, 53: 236-240).

[Analysis of olfactory dysfunction after endoscopic and microscopic endonasal transsphenoidal surgery for pituitary adenoma].

Objective:To analyze the clinical data of patients with olfactory dysfunction after endoscopic or microscopic endonasal transsphenoidal approach,and to screen out the possible related factors,to guide clinical diagnosis and treatment,and to improve the patients' quality of life.Method:In the retrospective study,we analyze patients' data,and follow up them with telephone,records the olfactory levels before and after surgery. In the prospective study,five odors were applied to test the olfactory function before the surgery,then one week,3 weeks and 6 weeks after that. The results were statistically analyzed.Result:In the retrospective study,olfactory dysfunction occurred in 67.74%,47.37%in MTS group and ETS group. The olfactory disorder had no significant difference (P>0.05) among MTS group and ETS group after surgery. And the analysis of factors showed no significant difference. In the prospective study,there is significant difference between the preoperative results and postoperative results about the operative side and nonoperative side in ETS group and MTS group (P<0.05). In all results,there is no significant difference between operative side and nonoperative side in ETS group and MTS group (P>0.05).Conclusion:There is olfactory dysfunction after transsphenoidal surgery to resect pituitary adenomas. To prepare the nasal condition well preoperatively,protect the nasal structure intraoperatively and aplicate appropriate treatment postoperatively may reduce the incidence of olfactory disorder.

[Role of sleep apnea monitoring management platform in the treatment of patients with obstructive sleep apnea hypopnea syndrome].

Objective:To investigate the clinical significance and value of the sleep apnea monitoring management platform in the treatment of patients with obstructive sleep apnea hypopnea syndrome (OSAHS) by comparing with the traditional continuous positive airway pressure (CPAP) card reader mode.Method:A total of 48 severe adult OSAHS patients from Department of Otorhinolaryngology Head and Neck Surgery-Sleep Medical Center of the Third People's Hospital of Honghe during the period of Nov. 2015 to Aug. 2016 were collected in this prospective study. All of them were diagnosed by PSG and treated with nasal continuous positive airway pressure. They were randomly divided into group A (n=24) and group B (n=24). Group A and B were treated with the traditional CPAP card reader mode and the sleep apnea monitoring management platform respectively. During the follow-up, the compliance, mean blood oxygen saturation, titration pressure, Epworth sleepiness scale after 1, 3, 6 and 12 month treatment were compared between two groups. Ttest was used to analyze the difference. Statistical significance was set at P <0.05.Result:Statistical analysis showed that there were significant difference in the compliance of using CPAP, mean blood oxygen saturation and Epworth sleepiness scale score between the two groups (P < 0.05), but the titration pressure had no difference between the two groups (P>0.05).Conclusion:Compared the sleep apnea monitoring management platform with traditional CPAP card reader mode in the treatment of OSAHS patients, the former could solve the problems during the CPAP use in time, improve the compliance of using CPAP, and which could increase the efficacy of CPAP. Thus, the sleep apnea monitoring management platform is of more clinical value and deserve promotion.

Evolution and heterogeneity of non-hereditary colorectal cancer revealed by single-cell exome sequencing.

Recently single-cell whole-exome sequencing (scWES) has deeply expanded and sharpened our knowledge of cancer evolution and subclonality. Herein, with scWES and matched bulk whole-exome sequencing (bulk WES) on two colorectal cancer (CRC) patients with normal or adenomatous polyps, we found that both the adenoma and cancer were of monoclonal origin, and both shared partial mutations in the same signaling pathways, but each showed a specific spectrum of heterogeneous somatic mutations. In addition, the adenoma and cancer further developed intratumor heterogeneity with the accumulation of nonrandom somatic mutations specifically in GPCR, PI3K-Akt and FGFR signaling pathways. We identified novel driver mutations that developed during adenoma and cancer evolution, particularly in OR1B1 (GPCR signaling pathway) for adenoma evolution, and LAMA1 (PI3K-Akt signaling pathway) and ADCY3 (FGFR signaling pathway) for CRC evolution. In summary, we demonstrated that both colorectal adenoma and CRC are monoclonal in origin, and the CRCs further diversified into different subclones with heterogeneous mutation profiles accumulating in GPCR, PI3K-Akt and FGFR signaling pathways. ScWES provides evidence for the importance of mutations in certain pathways that would not be as apparent from bulk sequencing of tumors, and can potentially establish whether specific mutations are mutually exclusive or occur sequentially in the same subclone of cells.

[Clinical research of anxiety and depression state among patients suffered from sudden deafness with vertigo].

Objective:To offer the objective evidences for the diagnosis by analyzing anxiety and depression among patients with sudden deafness and vertigo and their possible causes. Method:Eighty patients with sudden deafness and vertigo of single ear as the observation group; 80 patients with sudden deafness without vertigo and 60 healthy people as control group.Hamilton anxiety scale (HAMA), Hamilton depression scale (HAMD) are used for quantitative analysis and compare the results of each group score. Result:①The HAMA score of the deafness group with vertigo was significantly higher than that of the group without vertigo and the healthy group (P<0.01). The HAMD score of the deafness group with vertigo was significantly higher than that of the group without vertigo and the healthy group (P<0.01). ②There was correlation between HAMA and sex, age, duration of dizziness, tinnitus, DHI and efficacy grading (P<0.05). There was no correlation between HAMA and the mean hearing threshold, urban and rural distribution, ear fullness (P> 0.05). There was a correlation between HAMD and sex, age, tinnitus, DHI and efficacy grading (P<0.05). There was no correlation between HAMD and the mean hearing threshold, duration of dizziness, urban and rural distribution, ear fullness (P>0.05). Conclusion:There are anxiety and depression in patients with deafness and vertigo. It is mainly related to sex, age, dizziness duration, tinnitus, DHI and grade of efficacy. It is necessary to pay attention to the mental state of patients in clinical treatment. Psychological counseling or treatment can be used to improve clinical efficacy.

GADD45B mediates podocyte injury in zebrafish by activating the ROS-GADD45B-p38 pathway.

GADD45 gene has been implicated in cell cycle arrest, cell survival or apoptosis in a cell type specific and context-dependent manner. Members of GADD45 gene family have been found differentially expressed in several podocyte injury models, but their roles in podocytes are unclear. Using an in vivo zebrafish model of inducible podocyte injury that we have previously established, we found that zebrafish orthologs of gadd45b were induced upon the induction of podocyte injury. Podocyte-specific overexpression of zebrafish gadd45b exacerbated edema, proteinuria and foot-process effacement, whereas knockdown of gadd45b by morpholino-oligos in zebrafish larvae ameliorated podocyte injury. We then explored the role of GADD45B induction in podocyte injury using in vitro podocyte culture. We confirmed that GADD45B was significantly upregulated during the early phase of podocyte injury in cultured human podocytes and that podocyte apoptosis induced by TGF-ß and puromycin aminonucleoside (PAN) was aggravated by GADD45B overexpression but ameliorated by shRNA-mediated GADD45B knockdown. We also showed that ROS inhibitor NAC suppressed PAN-induced GADD45B expression and subsequent activation of p38 MAPK pathway in podocytes and that inhibition of GADD45B diminished PAN-induced p38 MAPK activation. Taken together, our findings demonstrated that GADD45B has an important role in podocyte injury and may be a therapeutic target for the management of podocyte injury in glomerular diseases.

A longitudinally split rabbit segmental gracilis to simulate penile erectile function: anatomic basis and animal models.

OBJECTIVE: The gracilis was once applied in reconstructing erectile function but its appearance was bulky. We aimed to design a model meeting the requirements of both reducing volume and retaining function. MATERIALS AND METHODS: The gracilis muscles of 6 rabbits were harvested, applied colorful vascular perfusion and modified Sihler's intramuscular nerve staining. According to their intramuscular nerves and blood vessels, 9 rabbit right gracilis muscles were then longitudinally split into two halves. The anterior muscle bundle was selected as the functional unit and blood supply. RESULTS: The intramuscular nerves and vessels were simultaneously presented on a same specimen. Their relationship suggested gracilis muscle to be composed of two relatively independent subunits. The reconstructed penis survived well, simulating erectile action satisfactorily. DISCUSSION: The penis model reconstructed with longitudinally split rabbit segmental gracilis myocutaneous flap had met the requirements of both restoring erectile function and improving the appearance.

The antibody-based CA125-targeted maintenance therapy for the epithelial ovarian cancer: a meta-analysis.

OBJECTIVES: To assess the effect and toxicity of CA125-targeted antibody used as maintenance therapy for advanced epithelial ovarian cancer (EOC). MATERIALS AND METHODS: Two reviewers searched PubMed, Medline, Embase, VIP databases, and the references of selected articles for randomized controlled trials comparing maintenance CA125-targeted antibody treatment with placebo/observation. One-, two-, three-, and five-year overall survival (OS) and progression free survival (PFS) were collected. Incidence and severity of adverse events were extracted. Meta-analysis of combined risk ratio (RR) for OS , PFS, and toxicity were conducted. RESULTS: Four trials including 1,259 women were identified. Meta-analysis showed the combined RR was 1.02 (95% CI, 0.85-1.22) for three-year OS and 0.98 (95% CI, 0.70-1.39) for the three-year PFS. This review found that abagovomab and oregovomab caused toxicity no more than placebo. CONCLUSIONS: CA125-targeted antibody used as maintenance therapy alone is not more effective than placebo but they were safe as maintenance therapy.

The construction of cDNA library and the screening of related antigen of ascitic tumor cells of ovarian cancer.

OBJECTIVE: To construct the cDNA library of the ascites tumor cells of ovarian cancer, which can be used to screen the related antigen for the early diagnosis of ovarian cancer and therapeutic targets of immune treatment. MATERIALS AND METHODS: Four cases of ovarian serous cystadenocarcinoma, two cases of ovarian mucinous cystadenocarcinoma, and two cases of ovarian endometrial carcinoma in patients with ascitic tumor cells which were used to construct the cDNA library. To screen the ovarian cancer antigen gene, evaluate the enzyme, and analyze nucleotide sequence, serological analysis of recombinant tumor cDNA expression libraries (SEREX) and suppression subtractive hybridization technique (SSH) techniques were utilized. The detection method of recombinant expression-based serological mini-arrays (SMARTA) was used to detect the ovarian cancer antigen and the positive reaction of 105 cases of ovarian cancer patients and 105 normal women's autoantibodies correspondingly in serum. RESULTS: After two rounds of serologic screening and glycosides sequencing analysis, 59 candidates of ovarian cancer antigen gene fragments were finally identified, which corresponded to 50 genes. They were then divided into six categories: (1) the homologous genes which related to the known ovarian cancer genes, such as BARD 1 gene, etc; (2) the homologous genes which were associated with other tumors, such as TM4SFI gene, etc; (3) the genes which were expressed in a special organization, such as ILF3, FXR1 gene, etc; (4) the genes which were the same with some protein genes of special function, such as TIZ, ClD gene; (5) the homologous genes which possessed the same source with embryonic genes, such as PKHD1 gene, etc; (6) the remaining genes were the unknown genes without the homologous sequence in the gene pool, such as OV-189 genes. CONCLUSION: SEREX technology combined with SSH method is an effective research strategy which can filter tumor antigen with high specific character; the corresponding autoantibodies of TM4SFl, ClD, TIZ, BARDI, FXRI, and OV-189 gene's recombinant antigen in serum can be regarded as the biomarkers which are used to diagnose ovarian cancer. The combination of multiple antigen detection can improve diagnostic efficiency.

Association of the status of interstitial cells of Cajal and electrogastrogram parameters, gastric emptying and symptoms in patients with gastroparesis.

Our goal was to investigate associations between the status of interstitial cells of Cajal (ICC) and electrogastrogram (EGG) parameters, gastric emptying and symptoms in a large cohort of patients with gastroparesis. Forty-one patients with refractory gastroparesis who were referred for gastric electrical stimulation (GES) underwent full thickness gastric (antrum) biopsy during the surgery to place the GES device. The biopsy samples were stained with c-kit and scored for the presence of ICC based on criteria obtained from 10 controls. All patients underwent EGG recordings, a 4-h standardized scintigraphic gastric emptying study and symptom assessment prior to the surgery. Based on antral biopsy, 15 patients (36%) had almost no ICC (ICC- group) and 26 patients had adequate cell numbers (ICC+ group). EGG recordings in the ICC- group displayed significantly less normal slow waves than in the ICC+ group both in the fasting and fed states. Tachygastria in the ICC- group was significantly more than in the ICC+ group both in the fasting (32 +/- 8%vs 11 +/- 2%) and fed states (27 +/- 9%vs 12 +/- 2%). There was no statistical difference in gastric emptying, symptom severity of gastroparesis, aetiology, age and gender between the two groups. Severely depleted ICC occurs in up to 36% of gastroparetic patients and significantly correlates with an abnormal EGG. Severely depleted ICC does not correlate with the severity of gastroparesis as assessed by gastric emptying or symptom status but did result in a poorer symptomatic response to GES. These data suggest that the EGG may have a role for predicting ICC status during clinical evaluation of gastroparetic patients.