[Establishment and preliminary application of organoids in ovarian cancer].

Objective: To explore the establishment and application of ovarian cancer organoids. Methods: Fresh ovarian tumor tissues, obtaining from patients underwent surgery in the First Affiliated Hospital of Nanjing Medical University between October 2021 and March 2022, were collected, enzymatic degraded, digested, and embedded into matrigel to establish organoids. A total of 32 ovarian cancer samples were collected. Hematoxylin eosin (HE) staining and immunofluorescence (IF) procedure were used to verify the morphological structure of organoids and their expression of molecular markers. 3D cyto-live or dead assay was used to detecte the live or dead cells in organoids. Carboplatin with a concentration ranging from 5 to 80 µmol/L (5, 10, 20, 40, 80 µmol/L) was added to organoids to calculate the 50% inhibitory concentration (IC50) in different organoids. Results: (1) Organoids from a total of 32 patients were established, of which 18 cases could be passaged stably in the long term in vitro, while 14 could be passaged in the short time. The average amplification time of long-term passage in vitro was over 3 months, and the longest reached 9 months. (2) In HE staining, significant nuclei atypia and local micropapillary structures were observed in organoids. IF staining revealed that ovarian cancer organoids expressed molecular markers similar to primary tumor tissues, such as Pan cytokeratin (Pan-CK), p53, paired box gene 8 (PAX8), and Wilms tumor gene 1 (WT1). (3) In 3D cyto-live or dead assay, a large number of apoptotic cells were observed inside and around the organoids after added carboplatin. The sensitivity to carboplatin varied in 18 organoids could amplify in the long term, with an average IC50 of (29.5±15.8) µmol/L. Moreover, IC50 values of 4 organoids derived from patients received neoadjuvant chemotherapy were much higher than the 14 organoids which did not received neoadjuvant chemotherapy [(48.7±11.3) µmol/L vs (24.0±12.1) µmol/L; t=3.429, P=0.022]. Conclusions: Organoids recapitulate ovarian cancers in vitro and could be stably passaged. Organoids derived from patients received neoadjuvant chemotherapy have higher resistance to carboplatin.

[Six cases of acute toilet cleaners].

The oral toilet cleaners poisoning can result in devastating gastrointestinal tract injuries with the risk of perforation and/or hemorrhage, and sometimes potentially fatal systemic complications and sequelas. It should be given positive treatment. In this paper, six cases of acute toilet cleaners poisoning were analyzed, and the clinical characteristics and treatment effect were summarized, so as to improve the understanding, diagnosis and treatment level of the disease.

[Oncologic outcomes of early stage cervical cancer performed operation by different laparoscopic surgical procedures: analysis of clinical data from mutiple centers].

Objective: To evaluate the oncologic outcomes of different laparoscopic radical hysterectomy. Methods: From January 2011 to December 2014, the laparoscopic operation cases of cervical cancer at stage Ⅰb1, Ⅰb2, Ⅱa1 and Ⅱa2, including the histologic subtypes of squamous-cell carcinoma, adenocarcinoma and adenosquamous carcinoma, were collected in five clinical centers. The data were divided into two groups according to the surgical procedures, that is, modified laparoscopic-vaginal radical hysterectomy (mLVRH) and total laparoscopic radical hysterectomy (TLRH). The overall survival rate (OS), disease-free survival rate (DFS) at 5 years were retrospectively analyzed in this study. Results: There were 674 cases in total, including 377 cases of mLVRH, 297 cases of TLRH. (1) The OS at 5 years: the mLVRH was 96.1% and the TLRH was 92.0%, and the mLVRH was higher than that of TLRH （P=0.010）. Stratify analysis， including stage of disease （Ⅰb1 and Ⅱa1）， histologic subtypes （squamous-cell carcinoma, adenocarcinoma）， lymph node metastasis， revealed that, ① Stage of disease: in stage Ⅰb1, the OS at five years of mLVRH was higher than that in TLRH group （98.6% vs 93.6%, P=0.012）. In stage Ⅱa1, there was significant difference between the two groups, the OS at five years of mLVRH and TLRH were 93.6% and 77.6% （P=0.007）. ② Histologic subtypes: for the OS at five years of squamous-cell carcinoma, mLVRH and TLRH were 96.1% and 92.3%, and there was significant difference （P=0.046）； for adenocarcinoma, the OS at five years were 91.0% and 88.6%， and there was no difference between two groups （P=0.230）. ③ Lymph node metastasis： the mLVRH and TLRH with lymph node metastasis, the OS at five years were 98.6% and 96.4%; the mLVRH and TLRH without lymph node metastasis, the OS at five years were 89.3% and 80.8%. There were no significant differences between the two groups,respectively (P=0.156， P=0.093）. (2) The DFS at 5 years: there was no significant difference between mLVRH and TLRH (94.1% vs 90.9%, P=0.220). Stratify analysis for stage of disease, the mLVRH group was higher than that in the TLRH group in stage Ⅰb1 (97.0% vs 92.8%, P=0.039). However, for stage Ⅱa1, there was no significant difference between mLVRH and TLRH group （88.2% vs 75.8%, P=0.074）. Conclusions: The results of this retrospective study indicated that different laparoscopy surgical procedures had diverse oncologic outcomes. The OS at 5 years of the mLVRH is superior to the TLRH. The DFS at 5 years in Ⅰb1 stage, the mLVRH is higher than the TLRH. Therefore, the modified laparoscopy is still an alternative surgery for early cervical cancer patients when following the principle of no-tumor-exposure.

[The attentional bias of emotion in medical workers of emergency department in one hospital and it's relationship with mood state].

Objective: To study the characteristics of attentional bias to emotional stimulus in medical workers of emergency department and explore the relationship between the emotional Stroop effect and mood states, thus providing a reference to the assessment and intervention of emergency medical workers' mental health. Methods: Two kinds of emotional Stroop tasks with words and pictures and the Profile of Mood States (POMS) were administered to thirty-six medical workers of emergency department and thirty-six medical workers of other departments. Results: The reaction time to negative and positive words and pictures were all higher than the reaction time to neutral stimulus, thus showing the significant Stroop effect in medical workers. The Stroop effect to negative emotional stimulus in medical workers of emergency department was significantly smaller than that in medical workers of other departments, while the difference of Stroop effect to positive stimulus was not significant between the two groups. Correlation analyses revealed that the correlations between Stroop effect size to positive pictures and tension and angry of POMS were all negatively significant (r value was -0.314 and -0.330 respectively) , and the correlation between Stroop effect size to negative pictures and fatigue was negatively significant (r=-0.303) . Conclusions: There is a significant emotional Stroop effect in medical workers. Although the ability of anti-interference to emotional stimulus in medical workers of emergency department was better than medical workers of the other departments, the long-term avoidance to negative emotional stimulus was still has a negative effect on the mood states of individuals.

Lowering the high rate of caesarean delivery in China: an experience from Shanghai.

OBJECTIVE: To examine the trends of caesarean delivery (CD) after an intervention to lower the high rate of CD at a Chinese maternity hospital. DESIGN: Retrospective cohort study. SETTING: A large tertiary obstetric centre in Shanghai, China, from 2007 to 2014. SAMPLE: 81 459 nulliparous women who delivered a term singleton infant. METHODS: Logistic regression was used to calculate the odds of CD while adjusting for confounders. MAIN OUTCOME MEASURE: Rate of CD before and after the intervention. RESULTS: The rate of CD decreased from 51.5% in 2008 to 36.1% in 2014, mostly due to a reduction in non-indicated antepartum CD from 27.9% in 2010 to 11.9% in 2014. After adjustment, a period effect remained with delivery between 2011 and 2014 associated with a 31% reduction in the odds of CD compared with delivery between 2007 and 2010 [odds ratio (OR): 0.69, 95% CI: 0.66-0.71)] and a 33% reduction in the odds of antepartum CD (OR: 0.67, 95% CI: 0.64-0.69). The frequencies of perinatal mortality (0.5 versus 0.4/1000), hypoxic ischaemic encephalopathy (0.9 versus 1.2/1000), meconium aspiration syndrome (0.5/1000), birth trauma (0.6/1000), respiratory distress syndrome (0.5% versus 0.4%) and necrotising enterocolitis (0.9 versus 0.6/1000) were similar. The frequency of neonatal infection increased slightly (0.6% versus 0.8%), although this could be explained by other factors. CONCLUSIONS: A marked reduction in CD has occurred at an urban tertiary care centre as a result of efforts to reduce the high rate of caesarean delivery. No notable differences in neonatal outcomes were observed. TWEETABLE ABSTRACT: High rates of caesarean delivery can be lowered without increases in neonatal morbidity and mortality.

Microsatellite markers reveal genetic divergence among wild and cultured populations of Chinese sucker Myxocyprinus asiaticus.

Studies of genetic diversity and genetic population structure are critical for the conservation and management of endangered species. The Chinese sucker Myxocyprinus asiaticus is a vulnerable monotypic species in China, which is at a risk of decline owing to fluctuations in effective population size and other demographic and environmental factors. We screened 11 microsatellite loci in 214 individuals to assess genetic differentiation in both wild and cultured populations. The single extant wild population had a higher number of alleles (13) than the cultured populations (average 7.3). High levels of genetic diversity, expressed as observed and expected heterozygosity (HO = 0.771, HE = 0.748, respectively), were found in both wild and cultured populations. We also report significant differentiation among wild and cultured populations (global FST = 0.023, P < 0.001). Both STRUCTURE analysis and neighbor-joining tree revealed three moderately divergent primary genetic clusters: the wild Yangtze population and the Sichuan population were each identified as an individual cluster, with the remaining populations clustered together. Twenty-two samples collected from the Yangtze River were assigned to the cultured population, demonstrating the efficacy of artificial propagation to avoid drastic reduction in the population size of M. asiaticus. These genetic data support the endangered status of the M. asiaticus and have implications for conservation management planning.

Utilization of International Association of Diabetes and Pregnancy Study Groups criteria vs. a two-step approach to screening for gestational diabetes mellitus in Chinese women with twin pregnancies.

AIM: To evaluate prevalence and pregnancy outcomes using the International Association of Diabetes and Pregnancy Study Groups (IADPSG) criteria and screening protocol vs. a standard two-step screening approach for gestational diabetes mellitus in Chinese twin pregnancies. METHODS: A retrospective cohort study for pregnancies during 2007-2013 was performed in a tertiary hospital in Shanghai, China. Data were abstracted from the medical records of twin pregnancies delivered at the hospital. During the period 2007-2011, this hospital used a two-step approach with a 50 g screening with a cut-off value of ≥ 7.8 mmol/l followed by a 100 g diagnostic oral glucose tolerance test (OGTT) utilizing Carpenter-Coustan criteria. In 2012-2013, the hospital switched to the IADPSG protocol of universal 75 g OGTT. RESULTS: Among 1461 twin pregnancies, 643 were screened utilizing IADPSG criteria and 818 using the two-step protocol. Gestational diabetes mellitus was diagnosed more frequently in the IADPSG group than in the two-step group [20.4% and 7.0%, respectively; adjusted odds ratio (aOR) = 3.22; 95% confidence interval (CI) = 2.30-4.52]. During the IADPSG period, the incidence of pre-eclampsia was 38% lower in non-gestational diabetes mellitus affected pregnancies compared with the two-step period (aOR = 0.62; 95% CI = 0.44-0.87). We observed no significant differences in most perinatal outcomes between the two groups. CONCLUSION: Compared with a standard two-step approach to screening and diagnosis, the IADPSG screening method resulted in a three-fold increase in the incidence of gestational diabetes mellitus in twin pregnancies, with a 38% lower risk of pre-eclampsia but no significant difference in most perinatal outcomes in non-gestational diabetes mellitus affected pregnancies.

Propofol induces apoptosis and inhibits the proliferation of rat embryonic neural stem cells via gamma-aminobutyric acid type A receptor.

We investigated the effect of propofol on the proliferation and viability of rat embryonic neural stem cells (rENSCs) and the potential mechanisms involved. rENSCs were isolated and cultured in vitro and treated with 1, 10, or 50 µM propofol, while the control group was treated with 0.1 µM dimethyl sulfoxide. The effect of propofol on the proliferation and viability of rENSCs was examined by proliferation and apoptosis assays. Real-time polymerase chain reaction was employed to analyze the mRNA expression of checkpoint kinase 1 (Chk1) and p53 in rENSCs exposed to propofol. Immunoprecipitation assay and western blotting analysis were performed to analyze the effect of propofol on Chk1 and p53 activity. The gamma-aminobutyric acid type A (GABAA) receptor antagonist securinine was added to the rENSCs before being treated with propofol to investigate the role of the GABAA receptor in propofol-triggered effects on rENSCs. rENSCs specifically expressing nestin protein were successfully isolated and cultured for experiments. The inhibitory effect of propofol on rENSCs increased dose-dependently. The percentage of apoptotic cells increased to 11.7% and the activity of Chk1 and p53 enhanced after treatment with 50 µM propofol. However, addition of securinine abrogated propofol-induced apoptosis and activation of Chk1. The GABAA receptor mediates propofol-induced apoptosis and proliferation inhibition of rENSCs, possibly by modulating the Chk1/p53 signaling pathway.

A rare cause of severe diarrhoea diagnosed by urine metabolic screening: aromatic L-amino acid decarboxylase deficiency.

A 15-year-old Chinese male with infantile-onset hypotonia, developmental delay, ptosis, and oculogyric episodes presented with a history of chronic diarrhoea since the age of 5 years. At presentation, he had an exacerbation of diarrhoeal symptoms resulting in dehydration and malnutrition with a concurrent severe chest infection. In view of his infantile-onset hypotonia, oculogyric crises, and protracted diarrhoea, an autonomic disturbance related to neurotransmitters was suspected. Urine organic acid profiling was compatible with aromatic L-amino acid decarboxylase deficiency. The diagnosis was confirmed based on cerebrospinal fluid analysis and genetic mutation analysis. The patient was treated with a combination of bromocriptine, selegiline, and pyridoxine; a satisfactory reduction in diarrhoea ensued. Our report highlights the importance of urine organic acid screening in infantile-onset hypotonia, especially when accompanied by oculogyric crises, and severe diarrhoea which could manifest as a result of autonomic disturbance.

ATP5PO levels regulate enteric nervous system development in zebrafish, linking Hirschsprung disease to Down Syndrome.

Hirschsprung disease (HSCR) is a complex genetic disorder characterized by the absence of enteric nervous system (ENS) in the distal region of the intestine. Down Syndrome (DS) patients have a >50-fold higher risk of developing HSCR than the general population, suggesting that overexpression of human chromosome 21 (Hsa21) genes contribute to HSCR etiology. However, identification of responsible genes remains challenging. Here, we describe a genetic screening of potential candidate genes located on Hsa21, using the zebrafish. Candidate genes were located in the DS-HSCR susceptibility region, expressed in the human intestine, were known potential biomarkers for DS prenatal diagnosis, and were present in the zebrafish genome. With this approach, four genes were selected: RCAN1, ITSN1, ATP5PO and SUMO3. However, only overexpression of ATP5PO, coding for a component of the mitochondrial ATPase, led to significant reduction of ENS cells. Paradoxically, in vitro studies showed that overexpression of ATP5PO led to a reduction of ATP5PO protein levels. Impaired neuronal differentiation and reduced mitochondrial ATP production, were also detected in vitro, after overexpression of ATP5PO in a neuroblastoma cell line. Finally, epistasis was observed between ATP5PO and ret, the most important HSCR gene. Taken together, our results identify ATP5PO as the gene responsible for the increased risk of HSCR in DS patients in particular if RET variants are also present, and show that a balanced expression of ATP5PO is required for normal ENS development.

Meta-analysis of the effect of whole-body vibration training on the improvement of limb function in patients with Parkinson's disease.

OBJECTIVE: This study aimed to systematically evaluate the effect of whole-body vibration (WBV) training on the improvement of limb function in patients with Parkinson's disease (PD). MATERIALS AND METHODS: An electronic search was performed on the databases, including PubMed, Web of Science, Scopus, SCI-hub, ScienceDirect, Embase, IEEE, Medline, Wiley, ClinicaIKey, CNKI, Wanfang, VIP database, Chinese Medical Association, and CBM Database from inception to May 2022 to collect randomized controlled studies on whole-body vibration training for patients with Parkinson's disease. Two researchers independently screened the literature, extracted the data, and evaluated the quality of the literature, then used ReviewManager 5.4 software for quantitative statistical analysis, including heterogeneity test, sensitivity analysis, risk of bias assessment, combined outcome index effect size and effect size inspection. RESULTS: A total of 9 studies were included in the meta-analysis, including 346 patients, 174 patients in the control group and 172 patients in the observation group. Meta-analysis results showed that, compared with conventional physical therapy or drug therapy alone, whole-body vibration reduced the Unified Parkinson's Disease Rating Scale (UPDRSIII) score and significantly improved the motor function of patients with Parkinson's disease [MD=-2.39, 95% CI (-4.41, -0.37), Z=2.14 (p=0.23)]. Moreover, whole-body vibration significantly improved the walking stability of Parkinson's patients [MD=-1.96, 95% CI (-2.71-1.21), Z=1.17 (p=0.03)]. However, its improvement in balance ability [MD=-0.06, 95% CI (-0.77, 0.65), Z=1.07 (p=0.19)] and daily living ability [MD=0.03, 95% CI (-1.68, 0.74), Z=0.24 (p=0.87)] of patients, it was not t statistically significant. CONCLUSIONS: Compared with conventional therapy, WBV has certain advantages in improving the balance function and gait performance of PD patients, but the effect on balance ability and daily living ability is not significant. Thus, more high-quality research is required for further verification.

Selective regulation of osteoblastic OPG and RANKL by dehydroepiandrosterone through activation of the estrogen receptor ß-mediated MAPK signaling pathway.

The aim of the work was to investigate the differential regulation by dehydroepiandrosterone (DHEA) of the osteoblastic production via the estrogen receptor beta (ER ß)-mediated signaling pathway. Having developed hMG63-ER ß cells and hMG63-shER ß cells, we analyzed the regulation by DHEA of human osteoblastic viability, the receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG), and the differential expression of ER ß, ER α, or p-ERK1/2 (extracellular signal-regulated kinase) in hMG63, hMG63-shER ß, and hMG63-ER ß cells pretreated with or without U0126, flutamide, and ICI 182780, followed by DHEA culture. When the level of ER ß was high, DHEA (10 - 7 mol/l) could effectively amplify the proliferation and inhibit the etoposide-induced apoptosis of hMG63 cells (p<0.01 and p<0.05, respectively), which was blocked by U0126. When the expression of ER ß was silenced, DHEA could not significantly improve the viability of hMG63. In the presence of ER ß, DHEA activated the pERK1/2-MAPK signaling pathway but not p38 and JNK. Besides, the regulation of p-ERK1/2 upon DHEA treatment was mainly modulated by ER ß instead of androgen receptor and ER α. The secretion of OPG was declined following the silence of ER ß (p<0.05). RANKL and ER α, however, were unaffected by culture with or without DHEA and U0126, regardless of the ER ß level. DHEA seems to act selectively on osteoblasts via the dominant ER ß receptor, which mediates amplified cell viability through the MAPK signaling pathway involving pERK1/2 and upregulates the production of OPG rather than RANKL.

Serum amyloid A is elevated in the serum of lung cancer patients with poor prognosis.

BACKGROUND: Lung cancer is known as the top cancer killer in most developed countries. However, there is currently no promising diagnostic or prognostic biomarker for lung cancer. This study aims to discover non-invasive differential markers in the serum of lung cancer patients, to determine the protein identity of the candidate biomarker(s), and to investigate any clinical implication of the biomarker(s) concerned. METHODS: Blood specimens were collected from 154 pre-operative patients with lung cancer and 35 healthy blood donors with no evidence of lung cancer. Fractionated serum samples were processed by surface-enhanced laser desorption/ionisation time-of-flight mass spectrometry (MS). Candidate biomarker was identified using sodium dodecyl sulphate polyacrylamide gel electrophoresis and tryptic digestion followed by tandem MS fragmentation analysis, which was subsequently validated with immunoassay. RESULTS: A differential protein with m/z 11.6 kDa was detected and identified as an isoform of human serum amyloid A (SAA). It was significantly increased by 1822% in lung cancer patients when compared with the healthy controls, which gave an area under the receiver operator characteristic curve of 0.88. In addition, the protein was also significantly elevated by 77% in lung cancer patients with survival <5 years when compared with patients with survival > or =5 years. CONCLUSION: There are several functions of the SAA protein, described in the context of inflammation, that are compatible with the mechanism of tumour invasion and metastasis. Our study not only detected increased SAA level in the serum of lung cancer patients but also identified that elevated SAA level may be a non-invasive biomarker useful for the prediction of lung cancer prognosis.

Expression and purification of two anti-CD19 single chain Fv fragments for targeting of liposomes to CD19-expressing cells.

Antibody-targeted liposomal anticancer drugs combine the specificity of antibodies with large payloads of entrapped drugs. We previously showed that liposomal doxorubicin (DXR) targeted via anti-CD19 monoclonal antibodies (mAb) or their Fab' fragments against the B-cell antigen CD19 led to improved therapeutic effects in murine B-cell lymphoma models relative to non-targeted liposomal DXR. We now are examining the use of anti-CD19 single chain fragments of the antibody variable region (scFv) as a targeting moiety, to test the hypothesis that scFv have advantages over full-sized mAb or Fab' fragments. We expressed two different anti-CD19 scFv constructs, HD37-C and HD37-CCH in E. coli, and purified the scFvs using two different methods. The HD37-CCH construct was selected for coupling studies due to its relative stability and activity in comparison to HD37-C. When coupled to liposomes, the HD37-CCH scFv showed increased binding in vitro to CD19-positive Raji cells, compared to non-targeted liposomes. Cytotoxicity data showed that HD37-CCH scFv-targeted liposomes loaded with DXR were more cytotoxic than non-targeted liposomal DXR. Our results suggest that anti-CD19 scFv constructs should be explored further for their potential in treating B-lymphoid leukemias and lymphomas.

Finite-temperature scaling of trace distance discord near criticality in spin diamond structure.

In this work we explore the quantum correlation quantified by trace distance discord as a measure to analyze the quantum critical behaviors in the Ising-XXZ diamond structure at finite temperatures. It is found that the first-order derivative of the trace distance discord exhibits a maximum around the critical point at finite temperatures. By analyzing the finite-temperature scaling behavior, we show that such a quantum correlation can detect exactly the quantum phase transitions from the entan-gled state in ferrimagnetic phase to an unentangled state in ferrimagnetic phase or to an unentangled state in ferromagnetic phase. The results also indicate that the above two kinds of transitions can be distinguished by the different finite-temperature scaling behaviors. Moreover, we find that the trace distance discord, in contrast to other typical quantum correlations (e.g., concurrence, quantum discord and Hellinger distance), may be more reliable to exactly spotlight the critical points of this model at finite temperatures under certain situations.