[Exploration of clinical pathway-oriented optimal management diagnosis and treatment model for rare diseases].

This study takes Cushing's syndrome, a rare disease, as a model, and adopts the path of "Plan, Do, Check, Action" (PDCA) to explore new methods to optimize the clinical path, can improve the quality and efficiency of diagnosis and treatment of rare diseases. After sorting out the problems existing in the previous diagnosis and treatment mode, our team optimizes the path in various ways and establishes a standard operation procedure (SOP) for the new path. In the evaluation of the optimized mode, 55 patients with Cushing's syndrome were admitted to the Department of Endocrinology, Peking Union Medical College Hospital, including 19 males and 36 females, aged (41.8±14.4) years (6-68 years). The pathway group (28 cases) and the control group (27 cases) were divided according to whether they were included in the new path management at the time of admission, and the effect of path optimization was assessed in terms of time, efficacy, safety and cost. The results showed that compared with the control group, the pathway group had a shorter time of hospitalization in the Department of Endocrinology and critical tests, such as blood cortisol rhythm, low-dose dexamethasone inhibition test, and bilateral inferior petrosal sinus sampling (all P<0.05). There was no significant differences in the decrease of total cortisol after operation, the incidence of postoperative complications, and hospitalization expenses (all P>0.05). The optimized path improves the medical efficiency while ensuring medical quality, safety and no increase in cost. This study proposes PDCA path optimization for complex diseases and establishes SOP process, which provides experience in management optimization for the patient-centered and clinical path-oriented diagnosis and treatment mode of rare diseases.

[Preliminary study on the relationship between biological rhythm changes and pregnancy complications during pregnancy].

To study the characteristics of heart rate rhythm in pregnant women at different trimester of pregnancy, and to explore the relationship between the basic rhythm of heart rate and pregnancy complications. Thirteen pregnant women who were diagnosed with normal early pregnancy in the Reproductive Center of the Second Affiliated Hospital of Soochow University from June 2018 to December 2019 were prospectively selected. Personal files were created and the heart rate data of pregnancy women was collected 24 hours a day by wearable devices until delivery. Prenatal examination and pregnancy outcomes were surveyed at follow-up. The cosine analysis method and the designed statistical module were used to analyze the long-term rhythm of pregnant women's heart rate. The heart rate of pregnant women showed a significant rhythm at different gestational weeks. Compared with the gestational week of 12, the midline-estimating statistic of rhythm(MESOR) increased significantly at the gestational week of 28 and 32 (t=-2.751,P=0.013;t=-2.314,P=0.032).The phase of rhythm shifted from 14∶00 pm in the first trimester of pregnancy (12 weeks) to 16∶00 pm in the second trimester (24 weeks) (t=2.613,P=0.018) and returned to 14∶00 pm at the third trimester (32 weeks) (t=-2.176,P=0.046). Season had no significant effect on the changes of MESOR, amplitude and phase of maternal heart rate in the first trimester (t=-0.356,P=0.729;t=-0.777,P=0.464;t=-0.434,P=0.673), while season had no significant effect on the changes of MESOR, amplitude and phase in the third trimester (t=-0.663,P=0.532;t=-0.209,P=0.841;t=0.625,P=0.592). The heart rate of one pregnant woman with natural delivery had rhythm disorder from the start of labor to delivery. The heart rate of one pregnant woman with premature rupture of membranes showed rhythm disorder before and after the rupture of membranes, and smaller amplitude. Rhythm disturbance may play a suggestive role in preterm delivery and labor initiation. In conclusion, pregnancy may cause changes in the internal heart rate rhythm. Maternal internal rhythm disturbance may occur when delivery or premature rupture of membranes occurs. The heart rate rhythm of pregnant women may be related to some common complications of pregnancy such as premature rupture of membranes.

[The regulatory activity of interleukin-35 on CD14+monocytes in patients with chronic heart failure].

Objective: To investigate the changes of interleukin-35 (IL-35) level and CD14+monocytes function in patients with chronic heart failure (CHF). Methods: A total of 74 patients with CHF who were hospitalized in the Department of Cardiology of the First Affiliated Hospital of Zhengzhou University between July 2018 and June 2019 as well as 29 healthy controls (HC) were continuously enrolled. 20 ml fasting anticoagulant peripheral blood was collected in the morning, and plasma was separated. IL-35 level was measured by ELISA. Peripheral CD14+monocytes were purified, and the IL-35 receptor subunits (IL-12Rß2 and gp130 mRNA) relative levels were semi-quantified by real-time PCR. CD14+monocytes were stimulated with IL-35, and were cultured in direct contact or indirect contact with human umbilical vein endothelial cells (HUVEC). Cytokines and granzyme B secretion in the supernatants was measured by ELISA. The percentage of HUVEC death was calculated by measuring lactate dehydrogenase level. The difference of the above indicators were compared between the CHF group and the HC group. Results: The age for the CHF group was (59.4±12.1) years, and 58.1% (43 cases) of them were males. The age for the HC group was (53.9±9.8) years, and 65.5% (19 cases) of them were males. The plasma IL-35 level was higher in the CHF group than the HC group ((22.89±7.58) mg/L vs (16.42±5.47) mg/L, P<0.001). The gp130 mRNA relative level was also higher in the CHF group than the HC group (1.07±0.19 vs 0.98±0.15, P=0.022). CD14+monocytes induced HUVEC death in the CHF group was lower in both direct contact and indirect contact culture system than the HC group (both P<0.001). The granzyme B secretion was also lower in the CHF group than the HC group (P<0.001). The CD14+monocytes induced HUVEC death was down-regulated in response to granzyme B inhibition (P=0.011). Both the CD14+monocytes induced HUVEC death and the granzyme B secretion were reduced in response to IL-35 stimulation (both P<0.001). Conclusion: CHF patients have the elevated IL-35 expression. IL-35 induces CD14+monocytes dysfunction via the inhibition of granzyme B secretion. This process promoted the progression of heart failure.

[Correlation between the primary tumor size of endometrial carcinoma and lymph node metastasis and recurrence].

Objective: To investigate the clinical significance of the primary tumor size in patients with endometrial carcinoma (EC). Methods: A total of 385 patients with EC admitted to Peking University People's Hospital from January 2006 to December 2016 with complete follow up data were selected, whose tumor size data before biopsy were retrospectively studied. Results: (1) The mean diameter of the primary tumor was (3.6±1.8) cm (range: 1-15 cm). And 48 cases were 0-<2 cm, 78 cases were 2-<3 cm, 92 cases were 3-<4 cm, 73 cases were 4-<5 cm, 94 cases were ≥5 cm. The diameter of the tumor was associated with age <60 years old, premenopause, CA125≥35 kU/L, non-parturition, poor differentiation, stage Ⅲ-Ⅳ, depth of myometrial infiltration ≥1/2, cervical interstitial involvement, adnexal metastasis and lymph node metastasis (all P<0.05), but not associated with body mass index, hypertension, diabetes mellitus, pathology, lymph-vascular space invasion (all P>0.05). (2) Among the 334 patients underwent lymphadenectomy, 45 (13.5%, 45/334) cases with lymph node metastasis were observed. Stratified analysis showed that lymph node metastasis and recurrence rate of patients with EC gradually increased with the increase of tumor size (P<0.05). Adopting 2, 3, 4 and 5 cm as cut-off values of tumor size, there were significant differences in the rate of lymph node metastasis and recurrence among them observed (P<0.05), except for lymph node metastasis rate and recurrence rate when the cut-off value was 2 cm (P>0.05). (3) An receiver operating characteristic (ROC) curve analysis showed that a tumor diameter of 4.25 cm was the cut-off prognostic value to predict lymph node metastasis and recurrence of EC. Conclusions: Tumor diameter is significantly correlated with lymph node metastasis and recurrence in patients with EC. Tumor size should be considered in determining the scope of surgery and adjuvant therapy.

[The modulatory activity of T cell immunoglobulin and mucin domain-containing protein 3 on T lymphocytes in patients with chronic heart failure].

Objective: To investigate the expression of T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) in patients with chronic heart failure and its modulatory activity on T lymphocytes. Methods: Eighty-six patients with chronic heart failure (CHF group) who were hospitalized in Department of Cardiology, the First Affiliated Hospital of Zhengzhou University between January and October 2018 were enrolled in the study. Meanwhile, thirty-two healthy controls (HC group) who received healthy examination were also selected. Peripheral blood mononuclear cells were isolated, and TIM-3 expression of CD4(+)and CD8(+)T cells was investigated by flow cytometry. CD4(+)and CD8(+)T cells were purified, and were stimulated by anti-TIM-3 antibody. Interferon-γ (IFN-γ), interleukin (IL)-4, IL-10, IL-35, IL-17, and IL-22 expressions in the supernatants of cultured CD4(+)T cells and tumor necrosis factor-α (TNF-α) and IFN-γ expressions in the supernatants of cultured CD8(+) T cells were measured by enzyme-linked immunosorbent assay. mRNA expressions of T-bet, GATA-3, FoxP3, and RORγt in CD4(+)T cells and perforin and granzyme B in CD8(+)T cells were semi-quantified by real-time PCR. Student t test or paired t test was used for comparisons between the two groups. Results: TIM-3(+) CD4(+) T cell percentage significantly increased in CHF group than that of HC group (3.47%±1.06% vs 0.92%±0.27%, P<0.001). TIM-3(+)CD8(+)T cell percentage also notably elevated in CHF group compared to HC group (6.12%±1.91% vs 1.77%±0.63%, P<0.001). CD4(+)T and CD8(+)T cells were dysfunctional in chronic heart failure. The levels of IFN-γ, IL-17, and IL-22 secreted by purified CD4(+) T cells significantly reduced in CHF group, while IL-10 and IL-35 expressions elevated in CHF group (all P<0.05). The relative mRNA expression levels of T-bet and RORγt in CD4(+) T cells remarkably decreased in CHF group than those of HC group (all P<0.01), while relative expression level of FoxP3 mRNA increased in CHF group (1.93±0.88 vs 0.97±0.28, P=0.031). The levels of TNF-α and IFN-γ produced by purified CD8(+)T cells notably reduced in CHF group than those of HC group (all P<0.05), and relative mRNA expression levels of perforin and granzyme B also decreased in CHF group (all P<0.05). The levels of anti-TIM-3 antibody stimulation-produced IFN-γ, IL-17, and IL-22 increased (all P<0.05) but IL-35 secretion reduced [(61±13) ng/ml vs (72±17) ng/ml, P=0.029] by CD4(+)T cells in CHF group. The relative mRNA expression levels of T-bet and RORγt also elevated in response to anti-TIM-3 antibody stimulation (all P<0.05). Anti-TIM-3 antibody stimulation promoted TNF-α and IFN-γ production by CD8(+)T cells in CHF group (all P<0.01). The relative mRNA expression levels of perforin and granzyme B also increased (all P<0.05). Conclusion: TIM-3 was increasingly expressed in T cells from patients with chronic heart failure, and might take part in the regulation of T cell dysfunction in chronic heart failure.

[Outcome of negative pressure wound therapy on diabetic foot ulcers: an observational study].

Objective: To compare the treatment effects between negative pressure wound therapy (NPWT) and traditional therapy in diabetic patients with foot ulcers. Methods: Diabetic patients with foot ulcers were selected from Endocrinology Ward between January 2015 and December 2016. All the patients were consecutively selected to receive NPWT in addition to the traditional therapy. Each patient receiving NPWT was individually matched with a patient receiving only traditional therapy and selected as the control group. Patients were controlled in both groups for age, sex and history of diabetes, hemoglobin A1c (HbA1c) level, and severity of the ulcer. Each patient received therapy for 3 months and was evaluated at the end of treatment. Treatment efficacy was evaluated by examining the lesion area before and after the therapy. Outcomes were compared between the two groups using the Chi square test. Results: A total of 62 patients were included in the study, with 31 cases in each group.Three months later, in the NPWT group, 26 patients had good therapeutic effects, 5 patients had poor efficacy, and 4 patients underwent the surgical treatment. While in the control group, only 18 patients had good therapeutic effects, 13 patients had poor efficacy, and 6 patients underwent surgical treatment. The effective percentage was significantly higher in NPWT group than that of the control group(P<0.05). Conclusion: NPWT can effectively improve the healing process of patients with diabetic foot ulcers in 3 months.

Surface air plasma-induced cell death and cytokine release of human keratinocytes in the context of psoriasis.

BACKGROUND: Cold atmospheric plasma (CAP) has shown promise for wound healing, although little is understood of the underpinning mechanisms. Little has been reported so far of its potential use in the treatment of immune-mediated diseases such as psoriasis. OBJECTIVES: To study CAP-induced cell death and cytokine release in human keratinocytes as a first assessment of possible CAP use for psoriasis. METHODS: Using a CAP generator free of energetic ions, we observed its effects on keratinocytes in terms of morphology, cell viability and apoptosis, intracellular and mitochondrial reactive oxygen species (ROS), lysosomal integrity and mitochondrial membrane potential; and on secretion and expression of eight cytokines at protein and gene levels. RESULTS: CAP-induced reduced cell viability, apoptotic death and production of intracellular and mitochondrial ROS in dose-dependent manner. Mitochondrial dysfunction and lysosomal leakage were found in CAP-treated cells. It also induced release of interleukin (IL)-6, IL-8, tumour necrosis factor (TNF)-α and vascular endothelial growth factor (VEGF), and enhanced the mRNA expression of IL-1ß, IL-6, IL-8, IL-10, TNF-α, interferon-γ and VEGF. By contrast, IL-12 declined monotonically. CONCLUSIONS: The results suggest that with appropriate control of its dose, physical plasma could induce cell death via apoptotic pathways and enable simultaneous reduction in IL-12. These effects may be used to suppress keratinocyte hyperproliferation and to target T-cell activation to control amplification of inflammation. This provides an initial basis for further studies of CAP as a potential therapeutic option for inflammatory and immune-related diseases in dermatology, including psoriasis.

Cloning and characterization of a novel betaine aldehyde dehydrogenase gene from Suaeda corniculata.

Glycine betaine is an important quaternary ammonium compound that is produced in response to several abiotic stresses in many organisms. The synthesis of glycine betaine requires the catalysis of betaine aldehyde dehydrogenase (BADH), which can convert betaine aldehyde into glycine betaine in plants, especially in halotolerant plants. In this study, we isolated the full-length cDNA of BADH from Suaeda corniculata (ScBADH) using reverse transcriptase-polymerase chain reaction and rapid amplification of cDNA ends. Next, we analyzed the expression profile of ScBADH using real-time PCR. The results showed that ScBADH expression was induced in the roots, stems, and leaves of S. corniculata seedlings under salt and drought stress. Next, ScBADH was overexpressed in Arabidopsis, resulting in the transgenic plants exhibiting enhanced tolerance over wild-type plants under salt and drought stress. We then analyzed the levels of glycine betaine and proline, as well as superoxide dismutase (SOD) activity, during salt stress in WT and transgenic Arabidopsis. The results indicated that overexpression of ScBADH produced more glycine betaine and proline, and increased SOD activity under NaCl treatment. Our results suggest that ScBADH might be a positive regulator in plants during the response to NaCl.

Expression of recombinant human anti-TNF-α scFv-Fc in Arabidopsis thaliana seeds.

Recombinant human anti-tumor necrosis factor (TNF)-α scFv-Fc was expressed in TKO mutant Arabidopsis thaliana seeds using plant-specific codons. Immunoblotting using a human IgG1 antibody detected the expression of anti-TNF-α proteins in plants. Results from qRT-PCR analysis demonstrated that the time of harvest significantly affected the protein yield and quality. Our results indicate that the Phaseolus vulgaris ß-phaseolin promoter directed anti-TNF-α scFv-Fc expression in A. thaliana seeds, with a maximum yield obtained at 20-days of development. Although the yield of anti-TNF-α scFv-Fc protein was not very high, accumulation of recombinant proteins in seeds is an attractive and simple method that can be used to purify biologically active anti-TNF-α scFv-Fc.

[Laparoscopic reoperation for recurred antireflux surgery of gastroesophageal reflux disease].

OBJECTIVE: To investigate the safety and effectiveness of laparoscopic reoperation for patients with gastroesophageal reflux disease (GERD) recurred form previous anti-reflux surgery. METHODS: Totally 19 patients received laparoscopic reoperation for symptomatic and anatomic recurred GERD in Department of Gastroesophageal Reflux Disease, Rocket Force General Hospital from January 2008 to September 2015 were retrospectively analyzed. There were 12 male and 7 female patients. The average reoperation age was (48±14) years, the average duration of reoperation from original ones was (43±38) months. The patients underwent preoperative barium, endoscopy, manometry and 24-hour pH studies. Laparoscopic hiatal hernia repair plus fundoplication was carried out for reoperation. Gastroesophageal reflux related symptoms (reflux, heartburn, chest pain, chough, wheezing, chest tightness and globus sensation) before and after surgery were compared by a questionnaire. The patients' medication consumption, complications and satisfaction of the reoperation were investigated as well. The repeated measures analysis of variance was used for statistical comparison of data preoperatively and postoperatively. RESULTS: No major complication and death occurred. Six cases (32%) had complications such as diarrhea, increased passing wind, flatulence, dysphagia and abdominal pain. The GERD related symptom score of reflux, heartburn, chest pain, chough, wheezing, chest tightness and globus sensation all significantly decreased (F: 25.0 to 56.7; P: 0.000 to 0.001) after the reoperation, with 68% good outcome of all the patients. After a follow-up of (33±22) months after reoperation, 1 case had partial recurrence at the 3(rd) month after reoperation. For all the patients, 12 cases felt very satisfied or satisfied with the reoperation. CONCLUSION: Laparoscopic reoperation is generally effective with acceptable morbidity rates for patients with esophageal and extraesophageal symptoms recurred form previous hiatal repair and (or) fundoplication.

rbcS SRS4 promoter from Glycine max and its expression activity in transgenic tobacco.

The regulatory region of the ribulose-1,5-bisphosphate carboxylase small subunit gene SRS4 from soybean (Glycine max) was cloned using TAIL-PCR and general PCR, and named the rbcS promoter. The promoter was fused with the GUS gene and introduced into Nicotiana tabacum via Agrobacterium-mediated leaf disk transformation. In 4-week-old transgenic tobacco plants, the highest GUS expression levels were observed in the leaves, GUS activity was 7.13- and 7.40-fold higher in leaves than in stems and roots, respectively. Moreover, GUS activity was stimulated by light. In conclusion, spatial and light regulation of the soybean rbcS promoter was observed in N. tabacum, thus illustrating a leaf-specific and light-induced promoter.

Meta-analysis of confocal laser endomicroscopy for the detection of colorectal neoplasia.

AIM: Confocal laser endomicroscopy (CLE) is a recently developed technique used to image colorectal neoplasia. Trials have shown varied results when it is compared with conventional colonoscopy. A meta-analysis was performed to determine the diagnostic accuracy of CLE in the detection of colorectal neoplasia. METHOD: A search was performed for studies assessing the accuracy of CLE in colorectal neoplasia. Studies comparing CLE diagnostic accuracy with conventional endoscopy in the detection of colorectal neoplasia were included. Exclusion criteria included case reports or case series, reviews, duplicate reports or insufficient data in the paper. Seventy-eight titles came up in the initial search and six studies were selected. These were subjected to a meta-analysis. In all, 284 patients with 1030 lesions were included. Each patient underwent conventional colonoscopy and CLE. Per-lesion sensitivity and specificity with 95% CI were calculated. RESULTS: In the individual studies, the sensitivity ranged from 33.3% to 100% and specificity from 71.6% to 99.4%. The weighted and total pooled result (random effects model) for sensitivity was 81% (95% CI 77-85) and for specificity was 88% (95% CI 85-90). The area under the weighted symmetric summary receiver operating curve was 0.9186. In the endoscope-based CLE subgroup, the sensitivity was 82% (95% CI 69-91) and specificity was 94% (95% CI 91-96). In the probe-based CLE subgroup, the sensitivity was 81% (95% CI 76-85) and the specificity was 75% (95% CI 69-81). CONCLUSION: CLE, using either the endoscope-based CLE or probe-based CLE technique, has high sensitivity and specificity. It could therefore be considered as an alternative endoscopic method to distinguish neoplastic from non-neoplastic lesions.

Does the use of tranexamic acid increase the risk of VTE in patients with hemoptysis?

OBJECTIVE: The aim of this study was to create a predictive nomogram that can accurately identify the risk factors of venous thromboembolism (VTE) in hospitalized patients exhibiting hemoptysis. PATIENTS AND METHODS: The present study gathered clinical and demographic data of 1,052 hospitalized patients with hemoptysis at Dongyang Hospital between January 2016 and January 2021 through the Lejiu database. The patients were categorized into two groups: the thrombotic event group (n=123) and the non-thrombotic event group (n=929), based on the presence of VTE before discharge. The study utilized univariable and multivariable logistic regression analyses to identify the independent risk factors for VTE, with the occurrence of thrombotic events serving as the dependent variable. Furthermore, a nomogram prediction model was formulated to verify the findings. RESULTS: In hospitalized patients with hemoptysis, the risk of VTE was found to be independently associated with the administration of tranexamic acid (TXA), the presence of D-dimer, and the Charlson Comorbidity Index (CCI) score (p<0.05). CONCLUSIONS: A nomogram model was constructed to evaluate the probability of VTE in patients hospitalized with hemoptysis. This model allows for the timely detection of early VTE warning signs, which may ultimately reduce its occurrence.

[Biological function and clinical significance of long non-coding RNA LINC00342 in head and neck squamous carcinoma].

Objective: To investigate the relationship between the long-non-coding RNA LINC00342 expression and the clinicopathological parameters of head and neck squamous cell carcinoma (HNSCC) and the biological function of LINC00342 in HNSCC cells. Methods: The expression level of LINC00342 in the HNSCC was analyzed using transcriptome sequencing data from TCGA (The Cancer Genome Atlas) database, and the expressions of LINC00342 in laryngeal squamous cell carcinoma tissues (LSCC) of 27 patients in the First Hospital of Shanxi Medical University were detected by transcriptome sequencing. The expression levels of LINC00342 in human embryonic lung diploid cells 2BS, HNSCC cell lines FD-LSC-1, CAL-27 and Detroit562 were determined by real-time quantitative polymerase chain reaction (qPCR). RNAi (RNA interference) was used for LINC00342 knockdown in HNSCC cell lines, and the changes of malignant phenotype in the tumor cells after LINC00342 knockdown were examined by cell counting kit-8 (CCK-8), colony formation, flow cytometry, transwell invasion and migration assays. Bioinformatics analysis was performed to construct a LINC00342-centered competing endogenous RNA (ceRNA) regulatory network, and GO (Gene Ontology) enrichment analysis was performed. Statistical analysis and graphing were performed using SPSS 25.0 software and GraphPad Prism 6 software. Results: Mean LINC00342 levels in HNSCC tissues and TCGA database were higher than that in normal control tissues, but with no significantly statistical difference (P=0.522). LINC00342 expression levels were positively correlated with cervical lymph node metastasis and pathological grade in patients with HNSCC, with higher expression in male patients than in female patients (P<0.05). Transcriptome sequencing analysis showed that mean expression level of LINC00342 in LSCC tissues of 27 patients was significantly higher than that in the paired adjacent normal mucosa tissues (t=1.56, P=0.036). LINC00342 expression was significantly upregulated in HNSCC cell lines FD-LSC-1, CAL-27 and Detroit562 (t-values of -12.17, -23.26 and -388.57, respectively; all P<0.001). Knockdown of LINC00342 by transfecting si-LINC00342-1 and si-LINC00342-2 inhibited HNSCC cell proliferation (t-values of 8.95 and 4.84, 2.70 and 5.55, 2.02 and 3.70, respectively), colony formation (t-values of 6.66 and 6.17, 7.38 and 11.65, 4.90 and 5.79, respectively), migration (t-values of 8.21 and 7.19, 5.76 and 6.46, 6.28 and 9.92, respectively) and invasion abilities (t-values of 9.29 and 10.25, 11.30 and 11.36, 8.02 and 8.66, respectively), but promoting apoptosis in cell lines FD-LSC-1 and CAL-27 (t-values of -2.21 and -5.83, -3.05 and -5.25 respectively) (all P-values<0.05). The LINC00342-centered ceRNA network consists of 10 downregulated microRNA and 647 upregulated mRNA nodes. GO analysis results indicated that LINC00342-regulated mRNAs were enriched in 22 biological processes, 32 molecular functions, and 12 cellular components. Conclusion: High level of LINC00342 is associated with the malignant progression of HNSCC. LINC00342 promotes the proliferation, migration, invasion, and antagonizes apoptosis of HNSCC cells, which serves as a potential molecular marker in HNSCC.

Risk factors for prolongation of ileostomy closure operation.

OBJECTIVE: Understanding factors for prolonged operative time is essential for surgery. This study aims at identifying the factors related to prolonged ileostomy closure operation time. PATIENTS AND METHODS: The data of 88 patients who underwent ileostomy reversal in the Department of Gastrointestinal Surgery of Jiaxing First Hospital between January 2018 and October 2021 were retrospectively analyzed. Prolonged operation time was defined as operative time >65 minutes. The Chi-square test was used to compare data between patients with normal operative time (≤65 minutes) and patients with prolonged operative time. Univariate and multivariate logistic regression analyses were performed to identify the factors associated with prolonged ileostomy closure operation time. RESULTS: Among the 88 patients (mean age: 62.78 years), median ileostomy retention time was 127.50 (range: 61.00-1,192.00) days. The operation time ranged from 35.00 minutes to 125.00 minutes. Prolonged ileostomy closure time occurred in 41 (46.6%) patients. In univariate analysis, the factors associated with prolonged operation time were body mass index (BMI) ≥25 kg/m2, previous history of abdominal surgery, and manual suture. History of serious complications after the primary operation was associated with shorter operation time. In multivariate analysis, the independent risk factors for prolonged operation time were BMI ≥25 kg/m2 (OR = 4.552, 95% CI: 1.369-15.136, p = 0.013), previous history of abdominal surgery (OR = 4.377, 95% CI: 1.394-13.739, p = 0.011), and manual suture (OR = 3.941, 95% CI: 1.181-13.154, p = 0.026). CONCLUSIONS: Overweight, previous history of abdominal surgery, and manual suture appear to be risk factors for prolonged operative time for ileostomy closure.

[Association between child care environment and childhood early development].

Objective: To investigate the factors related to child care environment and the association between child care environment and children's early development. Methods: Using stratified cluster random sampling, a total of 22 509 children newly enrolled to kindergarten from 187 kindergartens of 16 districts in Shanghai in 2017 were enrolled. A survey was conducted by parent-reported questionnaire. The index of child care environment (ICCE) and the early human capacity index were used to evaluate family child care environment and children's early development respectively. The sample was divided into four groups based on the ICCE score: the lowest family childcare environment (ICCE≤10 scores), lower middle (ICCE=11 scores), upper middle (ICCE=12 scores), and the highest (ICCE=13 scores). The linear regression model and Logistic regression model were used to analyze the factors related to child care environment and the association between child care environment and children's early development respectively. Results: A total of 22 067 children aged (44±7) months, including 11 425 boys (51.8%) and 10 642 girls (48.2%), participated in this study. The multnomial linear regression revealed girls, higher maternal education, higher household annual income, single-child, non-divorced parents, and early breastfeeding experience were protective factors of child care environment for children newly enrolled to kindergarten (ß=0.064, 0.238, 0.119, 0.096, 0.113, and 0.032; 95% confidence interval (CI): 0.020-0.108, 0.175-0.302, 0.058-0.180, 0.046-0.146, -0.012-0.242, and -0.051-0.116; all P<0.01). The multinomial Logistic regression revealed compared with the highest child care environment group, the odds ratio of children's early development risks in upper midclle, lower middle, and the lowest child care environment groups were 1.543 (95%CI: 1.373-1.735, P<0.01), 2.537 (95%CI: 2.254-2.856, P<0.01), and 4.198 (95%CI: 3.757-4.690, P<0.01), respectively. Conclusions: The child care environment is not only significantly related to family structure and socioeconomic status, but also to early breastfeeding experience. The child care environment plays an important role in promoting childhood early development.

Dietary supplementation of methionine mitigates oxidative stress in broilers under high stocking density.

We aimed to investigate whether dietary supplementation of methionine could mitigate intestinal oxidative injury in broilers under high stocking density (HSD). In the grower phase (d 22-42), 576 broilers with similar body weight were randomly chosen and divided into 8 groups in a 2 × 4 factorial experiment. Two different stocking densities (14 and 20 broilers per m2) were tested with 4 different methionine levels: 0.35%, 0.4%, 0.45%, or 0.5%. Intestinal morphological and oxidative stress markers were assessed at the end of the test period. The results showed that mortality of broilers was significantly higher in the HSD group fed 0.35% methionine diet than the other groups, which was reversed by supplementation with 0.40% to 0.50% methionine. HSD significantly decreased feed intake and daily weight gain. HSD treatment significantly decreased T-AOC, activity of GPX (P < 0.01) and increased the level of PCO (P < 0.01), MDA (P = 0.052) of plasma. The decreased glutathione peroxidase activity in the liver and jejunum caused by HSD was alleviated by additional methionine. Supplementation of methionine increased the ration of GSH/GSSG in the plasma. The jejunum villus height and ratio of villus height to crypt depth under low stocking density conditions with 0.40% methionine diet were the highest, whereas the 0.45% methionine group was the highest under HSD conditions. Thus, additional dietary supplementation of methionine mitigates oxidative stress in broilers under HSD conditions and 0.40% to 0.45% methionine can be applied in cage rearing broiler production for amelioration of oxidative stress caused by HSD.

Low levels of organic compound trace elements improve the eggshell quality, antioxidant capacity, immune function, and mineral deposition of aged laying hens.

In the egg production industry, trace elements are required as additional dietary supplements to play vital roles in performance and egg quality. Compared to inorganic microelements (ITs), appropriate dose of organic trace microelements (OTs) are environmentally friendly and sufficient to satisfy the needs of hens. In order to evaluate the extent to which low-dose OTs replace whole ITs, the effects of organic copper, zinc, manganese, and iron compound on the performance, eggshell quality, antioxidant capacity, immune function, and mineral deposition of old laying hens were investigated. A total of 1 080 57-week-old Jing Hong laying hens were assigned to five groups with six replicates of 36 layers each for an 8-week experimental period. The birds were fed either a basal diet (control treatment (CT)) or the basal diet supplemented with commercial levels of inorganic trace elements (IT 100%) or the equivalent organic trace elements at 20%, 30%, and 50% of the inorganic elements (OT 20%, OT 30%, and OT 50%, respectively). Results showed that compared with those in the CT treatment, feeding hens with inorganic or organic microelement diet had significant effects on the eggshell quality, antioxidant capacity, immune function, and mineral deposition of old laying hens (P < 0.05). The eggshell strength and ratio between OT 30%, OT 50%, and IT 100% were similar at weeks 4 and 8, and the eggshell thickness of these groups was also similar at weeks 6 and 8. At week 8, the eggshell colour in OT 50% was darker than that in IT 100%. The mineral content in the eggshells of OT 50% and IT 100% significantly increased (P < 0.001), with no significant difference in effective thickness, mammillary thickness, and mammillary knob width between groups. There were no differences in the malondialdehyde content, total antioxidant capacity, and total superoxide dismutase activity in serum between OT 30%, OT 50%, and IT100%. While the catalase activities, the interleukin-1ß, interleukin-10, immunoglobulin G, and immunoglobulin M concentrations in serum were not significantly different between OT 50% and IT 100%. The mineral contents in the faeces of the organic groups were considerably reduced compared with those in IT 100% (P < 0.001). In conclusion, dietary supplementation with 30-50% organic compound microelements has the potential to replace 100% inorganic microelements in the hen industry for improving eggshell quality, mineral deposition in the eggshell, antioxidant capacity, and immune function, and reducing emissions to the environment without negative effects on laying performance.

MiR-200b-5p inhibits proliferation of ovarian cancer cells by targeting ATAD2 and regulating PI3K/AKT signaling pathway.

OBJECTIVE: The purpose of this study was to investigate the influences of micro ribonucleic acid (miR)-200b-5p on proliferation and apoptosis of ovarian cancer (OC) cells, and to explore its correlations with the target gene ATPase family, AAA domain containing 2 (ATAD2), and the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. MATERIALS AND METHODS: Human ovarian fibroblasts (HOFs) or human OC cell lines (A2780) were cultured in vitro, and then, A2780 cells were separately transfected with miR-200b mimics or miR-NC or cultured with ATAD2-specific inhibitor BAY-850. Thereafter, the expression levels of miR-200b and ATAD2 messenger RNA (mRNA) were measured via qRT-PCR, and the proliferative capacity of cells was detected by CCK-8 assay. Next, the cell apoptosis was determined by means of flow cytometry and one-step TUNEL assay. Finally, the targeted regulatory relationship between miR-200b and ATAD2 was examined using a Luciferase reporter assay system, and the protein expressions were detected through Western blot (WB) assay. RESULTS: It was found that the expression level of miR-200b was remarkably lower (p<0.05), while the mRNA expression level of ATAD2 was notably higher (p<0.05) in A2780 cells than those in HOFs. The transfection with miR-200b mimics markedly reduced the mRNA expression level of ATAD2 (p<0.05) and the proliferative capacity (p<0.05) and increased the apoptosis rate (p<0.05) of A2780 cells. Besides, it was detected via the Luciferase reporter assay system that miR-200b inhibited ATAD2. BAY-850 significantly decreased the expression level of ATAD2 protein (p<0.05) and the proliferative capacity (p<0.05) but improved the apoptosis rate (p<0.05) of cells. Moreover, both miR-200b mimics and BAY-850 could distinctly repress the protein expression levels of PI3K and p-Akt of the PI3K/Akt signaling pathway (p<0.05) and enhance the expression of suppressor gene p53 (p<0.05). CONCLUSIONS: MiR-200b-5p can inhibit the proliferation and promote the apoptosis of OC cells through targeted inhibition of ATAD2 expression and regulation of the PI3K/Akt signaling pathway.

[Study on the relationship between nasal and eye symptoms and psychological status in patients with allergic rhinitis].

Objective:The aim of this study is to analysis the correlation between nasal and eye symptom and psychological status in patients with allergic rhinitis, in order to provide a basis for improving the mental health of patients. Method:The nasal symptom score scale, eye symptom score scale and Kessler 10 were used in the study, and the survey data was analyzed. Result:At least one of 147 patients had a nasal symptom at a rate of 100% (147/147); at least one ocular symptom was 86.4% (127/147). Of the 147 patients, 29 had K10 scores more than 21.The nasal symptom including nasal congestion, itching, eye pain, eye swelling, tearing positively correlated with K10 total score. Conclusion:The clinical staffs must pay attention to the mental health status of AR patients, strengthen the assessment of nasal symptoms, eye symptoms and mental status, and give targeted intervention to patients with poor mental health.