[Long-term outcomes of childhood steroid-sensitive nephrotic syndrome].

Objective: To investigate the long-term outcomes and risk factors in children with steroid-sensitive nephrotic syndrome (SSNS). Methods: A retrospective cohort study was conducted on newly onset SSNS admitted to the Department of Pediatrics of the First Affiliated Hospital of Sun Yat-sen University from January 2006 to December 2010 and 105 cases with follow-up for more than 10 years were included. Clinical data including general characteristics, clinical manifestation, laboratory tests, treatment and prognosis. The primary outcome was the clinical cure, and the secondary outcomes were relapse or ongoing immunosuppressive treatment within the last 1 year of follow-up and complications at the last follow-up. According to the primary outcome, the patients were divided into clinical cured group and uncured group. Categorical variables were compared between 2 groups using the χ2 or Fisher exact test, and continuous variables by t or Mann-Whitney U test. Multiple Logistic regression models were used for multivariate analysis. Results: Of the 105 children with SSNS, the age of onset was 3.0 (2.1, 5.0) years, and 82 (78.1%) were boys, 23(21.9%) were girls. The follow-up time was (13.1±1.4) years; 38 patients (36.2%) had frequently relapsing or steroid-dependent nephrotic syndrome (FRNS or SDNS) and no death or progression to end-stage kidney disease. Eighty-eight patients (83.8%) were clinically cured. Seventeen patients (16.2%) did not reach the clinical cure criteria, and 14 patients (13.3%) had relapsed or ongoing immunosuppressive treatment within the last year of follow-up. The proportion of FRNS or SDNS (12/17 vs. 29.5% (26/88), χ2=10.39), the proportion of treatment with second-line immunosuppressive therapy (13/17 vs. 18.2% (16/88), χ2=21.39), and the level of apolipoprotein A1 at onset ((2.0±0.5) vs. (1.7±0.6) g/L, t=2.02) in the uncured group were higher than those in the clinical cured group (all P<0.05). Multivariate Logistic regression analysis showed that patients treated with immunosuppressive therapy had an increased risk of not reaching clinical cure in the long term (OR=14.63, 95%CI 4.21-50.78, P<0.001). Of the 55 clinically cured patients who had relapsed, 48 patients (87.3%) did not relapse after 12 years of age. The age at last follow-up was 16.4 (14.6, 18.9) years, and 34 patients (32.4%) were ≥18 years of age. Among the 34 patients who had reached adulthood, 5 patients (14.7%) still relapsed or ongoing immunosuppressive treatment within the last year of follow-up. At the last follow-up, among the 105 patients, 13 still had long-term complications, and 8 patients were FRNS or SDNS. The proportion of FRNS or SDNS patients with short stature, obesity, cataracts, and osteoporotic bone fracture was 10.5% (4/38), 7.9% (3/38), 5.3% (2/38), and 2.6% (1/38), respectively. Conclusions: The majority of SSNS children were clinically cured, indicating a favorable long-term prognosis. History of treatment with second-line immunosuppressive therapy was the independent risk factor for patients not reaching the clinical cure criteria in the long term. While it is not uncommon for children with SSNS to persist into adulthood. The prevention and control of long-term complications of FRNS or SDNS patients should be strengthened.

[Pathological evaluation of endoscopic submucosal dissection for early gastric cancer and precancerous lesion in 411 cases].

OBJECTIVE: To evaluate the pathological characteristics of endoscopic submucosal dissection (ESD) specimens for early gastric cancer and precancerous lesions, accumulating experience for clinical management and pathological analysis. METHODS: A total of 411 cases of early gastric cancer or precancerous lesions underwent ESD. According to the Japanese guidelines for ESD treatment of early gastric cancer and classification of gastric carcinoma, the clinicopathological data, pathologic evaluation, concordance rate of pathological diagnosis between preoperative endoscopic forceps biopsies and their ESD specimens (in 400 cases), as well as the risk factors of non-curative resection of early gastric cancer, were analyzed retrospectively. RESULTS: 23.4% (96/411) of the 411 cases were adenoma/low-grade dysplasia and 76.6% (315/411) were early gastric cancer. The latter included 28.0% (115/411) non-invasive carcinoma/high-grade dysplasia and 48.7% (200/411) invasive carcinoma. The concordance rate of pathological diagnosis between endoscopic forceps biopsies and ESD specimens was 66.0% (264/400), correlating with pathological diagnosis and lesion location (P < 0.01). The rate of upgraded diagnosis and downgraded diagnosis after ESD was 29.8% (119/400) and 4.2% (17/400), respectively. Among the 315 cases of early gastric cancer, there were 277 cases (87.9%) of differentiated type and 38 cases (12.1%) of undifferentiated type. In the study, 262 cases (83.2%) met with absolute indication, while 53 cases (16.8%) met relative indication. En bloc and curative resection rates were 98.1% and 82.9%, respectively. Risk factors for non-curative resection included a long diameter >20 mm (OR=3.631, 95%CI: 1.170-11.270, P=0.026), tumor infiltration into submucosa (OR=69.761, 95%CI: 21.033-231.376, P < 0.001)and undifferentiated tumor histology (OR=16.950, 95%CI: 4.585-62.664, P < 0.001). CONCLUSION: Several subjective and objective factors, such as the limitations of biopsy samples, the characteristics and distribution of the lesions, different pathological understanding, and the endoscopic sampling and observation, can lead to the differences between the preoperative and postoperative pathological diagnosis of ESD. In particular, the pathological upgrade of postoperative diagnosis was more significant and should receive more attention by endoscopists and pathologists. The curative resection rate of early gastric cancer in ESD was high. Non-curative resection was related to the long diameter, the depth of tumor invasion and histological classification. ESD can also be performed in undifferentiated early gastric cancer if meeting the indication criteria. The comprehensive and standardized pathological analysis of ESD specimens is clinically important to evaluate the curative effect of ESD operation and patient outcomes.

Virulence assessment of four Glaesserella parasuis strains isolated in Liaoning province of China.

Glaesserella parasuis (G. parasuis) is a part of the normal upper respiratory microbiota of healthy swine. In many studies, the serovars 1, 4, 5, and 12 of G. parasuis are considered to be highly virulent and its serovars 3, 6, 7, 9, and 11 are considered to be non-virulent. Until now, researchers have found that non-virulent strains of G. parasuis cause an increasing number of diseases. However, little is known concerning why non-virulent strains cause disease with the virulence changes. In present study, four G. parasuis strains were evaluated for their cytotoxicity property, which aims to compare their virulence. The results showed that highly virulent strains XX0306 and CY1201, as well as, non-virulent strains HLD0115 and YK1603 caused a series of pathological changes, increased lactate dehydrogenase (LDH) release, and decreased cell activity. In addition, compared to the control group, both highly and non-virulent strains showed similar trends, demonstrating that the method of classifying the virulence of G. parasuis based on its serovar is worth further deliberation. Hence, we investigated the adhesion capacity and invasion rate of G. parasuis, the results indicated that XX0306 and HLD0115 had the strongest adhesion and invasion ability, which contradicts the classification of the virulence of G. parasuis based on its serovar. The apoptosis degree induced by highly virulent strains was more intensive than non-virulent strains, as measured by annexin V and propidium iodide (PI) double staining. Through testing the expression of apoptosis-related genes Bcl-2 and Bax, we found highly virulent strains induced apoptosis by inhibiting the expression of Bcl-2.

Transcriptomics analysis reveals key lncRNAs and genes related to the infection of porcine lung macrophages by Glaesserella parasuis.

Glaesserella parasuis (G. parasuis) is the pathogen of Glässer's disease in pig herds, which can cause severe inflammatory responses. However, at present, the pathogenic mechanism of G. parasuis is not very clear. LncRNAs can regulate the expression of mRNA in a variety of ways, thereby causing host cells to produce a variety of functional changes in response to bacterial infection. Here, we detected the changes in lncRNAs and mRNAs of 3D4/21 cells after G. parasuis CY1201 strain (serotype 13) infection. A total of 876 lncRNAs and 2166 mRNAs were differentially expression in 3D4/21 cells after G. parasuis infection. GO and KEGG enrichment analysis showed that the differentially up-regulated lncRNA target genes were mainly involved in the response to extracellular stimuli, cell receptor signaling pathways and chemokine signaling pathways. The differentially down-regulated lncRNA target genes were mainly involved in ERK1/ERK2 cascade reaction and adhesion junctions. 44 lncRNAs were screened that might be related in inflammation. CeRNA regulatory network of the top five difference inflammation-related lncRNAs showed that the up-regulated lncRNA group involved 5 lncRNAs, 50 miRNAs and 49 mRNAs. Meanwhile, there were 26 miRNAs and 36 mRNAs in the top five down-regulated lncRNA group. Our results contribute to understand the basic role of lncRNAs in 3D4/21 cells during G. parasuis infection, and lay the foundation for following research.

The effect of air temperature, velocity and humidity on respiration rate and rectal temperature as an expression of heat stress in gestating sows.

Global warming combined with increased production (i.e. more piglets, more milk and consequently more heat) means that sows are more often challenged by heat stress. The objective was to develop an effective temperature (ET) equation to predict how air temperature, velocity and humidity affect the respiration rate (RR), rectal temperature (RT) and skin temperature (ST) as an expression of heat stress in gestating sows in order to elucidate the relationship between the thermal parameters and the sows' perception of the environment. The experimental room was equipped with a negative pressure ventilation system with diffuse air inlet through the ceiling, electrical heaters, steam generators and dehumidifiers. An air distribution unit was constructed to generate vertical air velocity. A total of 16 gestating sows were exposed to three temperatures (25°C, 29°C and 33°C), two levels of relative humidity (30% and 70%) and three levels of air velocity (0.2 ms-1, 1 ms-1 and 2.5 ms-1). The RR, RT and ST were recorded every 30 min throughout the three 2-h test periods. The estimated effects of humidity and velocity in relation to effect of temperature was nearly independent of whether it was determined from RR or RT, whereas the effect of humidity was much smaller when determined from ST. High coefficients of determination (>0.97) were found for the second order relationship between the estimated ET and RR, RT and ST. An increase in relative humidity from 50 to 70% corresponded to an increase in ET of 0.9°C, while an increase in air velocity from 0.2 to 1.0 ms-1 corresponded to a decrease in ET of 1.2°C. The applied ET equation was useful for expressing the combined effect of temperature, humidity and velocity on animals exposed to heat stress. However, multiplying the effect of velocity by the temperature gradient between the animal and the surrounding air did not improve the estimation.

LncRNA-599547 contributes the inductive property of dermal papilla cells in cashmere goat through miR-15b-5p/Wnt10b axis.

The lncRNA-599547 (619-nt in length) is identified in secondary hair follicle (SHF) of cashmere goat, but its functional roles in regulating the inductive property of dermal papilla cells (DPCs) remains unknown. We found that lncRNA-599547 had significantly higher expression in dermal papilla of cashmere goat SHF at anagen than its counterpart at telogen. The overexpression of lncRNA-599547 led to a significant increase of ALP and LEF1 expression in DPCs (p < 0.05), whereas, the siLncRNA-1 mediated silencing of lncRNA-599547 significantly down-regulated the expression of ALP and LEF1 in DPCs (p < 0.05). Based on biotin-labeled RNA pull-down assay, we found that lncRNA-599547 directly interacted with chi-miR-15b-5p in DPCs. Based on both overexpression and silencing analysis of lncRNA-599547, our results indicate that lncRNA-599547 promotes the expression of Wnt10b in DPCs but without modulating its promoter methylation level. Using the mRNA-3'UTR fragments of goat Wnt10b containing the predicted binding sites of chi-miR-15b-5p in Dual-luciferase Reporter Assays, we show that lncRNA-599547 modulates the expression of Wnt10b at the chi-miR-15b-5p mediated post-transcriptional level. Taken together, our results indicate that lncRNA-599547 sponges miR-15b-5p to positively regulate the expression of Wnt10 gene, and thereby contributes the inductive property of DPCs in cashmere goat.

[Treatment effectiveness and long-term prognosis of childhood-onset lupus nephritis].

Objectives: To analyze the clinical characteristics, treatment effectiveness and long-term prognosis of childhood-onset lupus nephritis (LN), and to explore the risk factors for progression to end-stage renal disease (ESRD). Methods: In this retrospective study, the clinical data including general conditions, clinical manifestations, laboratory examinations, treatment, following up (till December 31st, 2020) and prognosis of 343 children with LN who were treated and followed up in the First Affiliated Hospital of Sun Yat-sen University from January 1, 2003 to December 31, 2019 were analyzed. Complete remission rates were compared between different pathological types according to renal biopsies and flare rates were compared between complete remission group and partial remission group according to the treatment effectiveness after 6 months of induction treatment. To investigate the risk factors of ESRD, the prognosis of flare and non-flare cases, and of cases with normal and elevated serum creatinine levels at baseline, was compared. Chi-squared tests were used for comparison between groups, and cumulative survival rate and renal survival rates were calculated by Kaplan-Meier survival analysis. Risk factors for ESRD were analyzed by COX regression model. Results: Among the 343 children, 68 were males (19.8%) and 275 were females (80.2%) with a median age of 13.0 (11.0, 16.0) years. Regarding the renal symptoms, 305 (88.9%) children had proteinuria and 245 (71.4%) had hematuria; while for extra-renal manifestations, 273 (79.6%) had anemia, 183 (53.4%) had rashes and 165 (48.1%) had fever. A total of 212 (61.8%) children had severely active SLE at initial presentation. After 6 months of induction treatment, the complete remission rate was 63.8% (219/343) and the partial remission rate was 27.1% (93/343). The complete remission rate was significantly higher in type Ⅰ and type Ⅱ LN compared to type Ⅳ LN (10/12 vs. 82/135 (60.7%), χ²=3.936, P=0.047). One hundred and ten children who achieved remission, including complete remission and partial remission, experienced renal flare with a flare rate of 35.3% and a mean time to flare was (43.2±28.4) months. There was no significant difference in flare rates between complete and partial remission group (36.1% (79/219) vs. 33.3% (31/93), χ²=3.394, P=0.065). The follow-up time of all the children was 60.4 (32.3, 100.9) months. During the follow-up period, 15 children died and the cumulative survival rates at 3, 5 and 10 years were 97.2%, 96.4% and 93.3%, respectively; 14 children progressed to ESRD and the cumulative renal survival rates at 3, 5, and 10 years were 99.2%, 97.1%, and 93.4%, respectively. COX multivariate analysis demonstrated that elevated serum creatinine at baseline, nephritic flare and nephrotic flare were independent risk factors for progression of ESRD (hazard ratio (HR)=3.575, 21.550 and 8.590, 95%CI 1.127-11.341, 2.394-194.027 and 1.042-70.823, P=0.031, 0.006, and 0.046, respectively). Conclusions: Children with LN are characterized by high SLE disease activity and multi-system involvement at onset. After 6 months of induction treatment, most of LN children could achieve clinical remission but some would experience renal flare. Nephritic flare, nephrotic flare and elevated serum creatinine at onset are independent risk factors for the progression of ESRD in children with LN.

[Diagnostic and predictive values of three criterias of acute kidney injury in children with chronic kidney disease].

Objective: To compare the consistency in diagnosing and staging acute kidney injury (AKI) in children with chronic kidney disease (CKD) according to three criterias. Methods: Children with CKD hospitalized in the First Affiliated Hospital of Sun Yat sen University from January 2013 to December 2019 were analyzed retrospectively. These patients underwent serum creatinine examination more than twice during hospitalization. The AKI diagnosis and staging were performed for each patient according to the 2007 pRIFLE, 2012 KDIGO and 2018 pROCK criteria respectively. All the children were followed up for 1 year after discharge through outpatient visit, re-hospitalization or online consultation. The clinical characteristics and prognosis of CKD children with or without AKI that were diagnosed by 3 criteria were compared. Analysis of variance and chi-squared tests were used for the comparison among groups. Concordance between the different diagnostic criteria was evaluated using Cohen's kappa coefficient. Result: A total of 2 551 children with CKD were included in this study, with an age of (8±4) years. There were 1 628 boys and 923 girls. Nephrotic syndrome was the most prevalent primary disease (55.4%), followed by lupus nephritis (11.2%) and purpura nephritis (8.2%). Among all stages of CKD, CKD category G1 was the most common type (2 146 cases, 84.1%), followed by CKD category G2 (221 cases, 8.7%). AKI occurence rates according to pRIFLE, KDIGO and pROCK criteria were 33.9% (866/2 551), 26.2%(669/2 551) and 19.5% (498/2 551) respectively (χ²=136.3,P<0.01). The diagnostic consistency within three criteria for AKI was high in children with CKD (κ=0.702), but AKI staging consistency was low (κ=0.329). Both the diagnosis and staging consistency of three AKI criteria were poor in children with CKD category G5 (all κ<0.400). The length of hospital stay (LOS), hospitalization costs, the occurence of intensive care unit (ICU) admission and in-hospital mortality were significantly higher in children with AKI diagnosed by different criteria (P<0.05). After 1-year follow-up, the repeated admission rate and CKD staging progress significantly increased in children with AKI (P<0.05). In children with baseline serum creatinine≥200 µmol/L, compared with children who did not experience AKI during hospitalization, the LOS and the hospitalization costs in children who were diagnosed AKI according to pRIFLE or pROCK criteria was significantly higher (P<0.05). However, there was no significant difference in the LOS and hospitalization costs between children with or without AKI who were diagnosed according to KDIGO criteria (all P>0.05). Conclusions: AKI diagnosed by all of the three criteria (pRIFLE, KDIGO and pROCK criteria) was associated with the poor prognosis in children with CKD. However, in those whose baseline serum creatinine≥ 200 µmol/L, AKI diagnosed by pRIFLE and pROCK criteria could better reflect the poor outcomes than by KDIGO criteria.

Application of mouse model for evaluation of recombinant LpxC and GmhA as novel antigenic vaccine candidates of Glaesserella parasuis serotype 13.

Glaesserella parasuis (G. parasuis) has been one of the bacteria affecting the large-scale swine industry. Lack of an effective vaccine has limited control of the disease, which has an effect on prevalence. In order to improve the cross-protection of vaccines, development on subunit vaccines has become a hot spot. In this study, we firstly cloned the lpxC and gmhA genes from G. parasuis serotype 13 isolates, and expressed and purified their proteins. The results showed that LpxC and GmhA can stimulate mice to produce IgG antibodies. Through testing the cytokine levels of interleukin 4 (IL-4), IL-10 and interferon-γ (IFN-γ), it is found that recombinant GmhA, the mixed LpxC and GmhA can stimulate the body to produce Th1 and Th2 immune responses, while recombinant LpxC and inactivated bacteria can only produce Th2 immune responses. On the protection rate for mice, recombinant LpxC, GmhA and the mixture of LpxC and GmhA can provide 50%, 50% and 60% protection for lethal dose of G. parasuis infection, respectively. The partial protection achieved by the recombinant LpxC and GmhA supports their potential as novel vaccine candidate antigens against G. parasuis.

LncRNA-MEG3 Regulates the Inflammatory Responses and Apoptosis in Porcine Alveolar Macrophages Infected with Haemophilus parasuis Through Modulating the miR-210/TLR4 Axis.

Haemophilus parasuis (H. parasuis, HPS) can elicit serious inflammatory responses and cause enormous economic loss to swine industry worldwide. However, the factors responsible for systemic infection and inflammatory responses of HPS have not yet been fully clarified. In this study, we found that lncRNA-MEG3 was significantly up-regulated in porcine alveolar macrophages (PAMs) infected with HPS. The gain- and loss-of-function analysis confirmed that lncRNA-MEG3 participated in the inflammatory responses and apoptosis in HPS-infected PAMs, which was assessed via several inflammatory cytokine genes (TNF-α, IL-1ß, and IL-6) and apoptotic factors (Bcl-2, Bax, and C-caspase-3). Based on biotin-labeled RNA pull-down assay, we found that lncRNA-MEG3 bound with miR-210 in HPS-infected PAMs. Based on both overexpression and knockdown analysis of lncRNA-MEG3, our results indicated that lncRNA-MEG3 promoted the expression of TLR4 in HPS-infected PAMs. Using dual-luciferase reporter assays, we showed that lncRNA-MEG3 positively regulated the expression of TLR4 gene in HPS-infected PAMs through miR-210 pathway. Taken together, our results indicated that lncRNA-MEG3 participated in the inflammatory responses and apoptosis in HPS-infected PAMs through modulating the miR-210/TLR4 axis. The results from this investigation provided significant information for a novel target to control HPS infection in swine.

Comparative Transcriptomic Analyses of Haemophilus parasuis Reveal Differently Expressed Genes among Strains with Different Virulence Degrees.

Haemophilus parasuis is commonly found in the upper respiratory tract of the pigs. Some isolates of H. parasuis can lead to both pneumonia and Glässer's disease of pigs with severe clinical symptoms. The virulence-associated genes for the various degrees of virulence observed in H. parasuis remains poorly understood. In the present study, we identified the differentially expressed genes between YK1603 (non-virulent strain) and XM1602 (moderately virulent strain) or CY1201 (highly virulent strain) of H. parasuis using Illumina sequencing technique. In comparison to YK1603, a total of 195 genes were significantly changed in CY1201, of which 71 genes were up-regulated and 124 genes were down-regulated, whereas 705 genes were significantly changed in XM1602, of which 415 genes were up-regulated and 290 genes were down-regulated. The enriched analysis of Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways on the differentially expressed genes showed that both enriched main GO terms and KEGG pathways appear to be different between the two kinds of comparision: CY1201 versus YK1603, and XM1602 versus YK1603. Based on real-time PCR technique, on the whole, it was confirmed that the expression of ten genes: lpxL, tbpB, kdtA, waaQ, oapA, napA, ptsH, mmsA, lpxM, and lpxB were agreement with the findings in Illumina sequencing analysis. These identified genes might participate in the regulation of a wide range of biological process involved in virulence of H. parasuis, such as phosphotransferase system and ABC transporters. Our results from this study provide a new way to gain insight into the virulent mechanisms of H. parasuis.

[Analysis of the self-conscious health status and influencing factors of greenhouse agricultural workers in Beijing suburb].

Objective: To explore the self-conscious health status and related influencing factors of greenhouse agricultural workers in Beijing suburb, so as to provide scientific basis for developing strategies to improve and promote the occupational health conditions of those workers. Methods: According to the production characteristics and scale of the main agricultural districts or counties in the suburb of Beijing, 182 agricultural production personnel were randomly selected to investigate the general situation, occupational situation and self-conscious health status during June 2018 to December 2019. The relevant factors which may affect the self-conscious health conditions were also analyzed by statistical methods. Results: The detection or reported rate of self-conscious health problems was 51.6% (94/182) , among which 29.1% (53/182) workers reported musculoskeletal disorder diseases and 21.4% (39/182) workers reported nervous system diseases. And the self-conscious health status of greenhouse agricultural workers were statistically different in household registration, gender, marital status, working years, mixing or spraying pesticides, smoking or eating in the greenhouse (P<0.05) . Moreover, gender, mixing or spraying pesticide, eating and smoking behavior in the workplace all had an impact on the risk of self-conscious health status of greenhouse agricultural workers (P< 0.05) . Specifically, male is the protective factor to reduce the occurrence of self-conscious symptoms of greenhouse agricultural workers (OR=0.447, 95%CI: 0.234~0.852) , while mixing or spraying pesticides and smoking or eating behaviors in the workplace are the risk factors (OR=1.055, 2.524; 95%CI: 0.503~2.210, 1.107~5.755) . Conclusion: Reducing pesticide use from the source thus minimize related exposure opportunities, strengthening occupational health education thus foster good hygienic habits and improve individual protection consciousness are of great significance for the health protection of greenhouse agricultural workers in Beijing suburb.

Full Chromosomal Relationships Between Populations and the Origin of Humans.

A comprehensive description of human genomes is essential for understanding human evolution and relationships between modern populations. However, most published literature focuses on local alignment comparison of several genes rather than the complete evolutionary record of individual genomes. Combining with data from the 1,000 Genomes Project, we successfully reconstructed 2,504 individual genomes and propose Divided Natural Vector method to analyze the distribution of nucleotides in the genomes. Comparisons based on autosomes, sex chromosomes and mitochondrial genomes reveal the genetic relationships between populations, and different inheritance pattern leads to different phylogenetic results. Results based on mitochondrial genomes confirm the "out-of-Africa" hypothesis and assert that humans, at least females, most likely originated in eastern Africa. The reconstructed genomes are stored on our server and can be further used for any genome-scale analysis of humans (http://yaulab.math.tsinghua.edu.cn/2022_1000genomesprojectdata/). This project provides the complete genomes of thousands of individuals and lays the groundwork for genome-level analyses of the genetic relationships between populations and the origin of humans.

[Characteristics of lymph node metastasis and evaluating the efficacy of endoscopic submucosal dissection in early gastric cancer].

OBJECTIVE: To investigate the correlation between clinicopathological features and lymph node metastasis, and to evaluate the feasibility and efficacy of endoscopic submucosal dissection (ESD) in early gastric cancer (EGC) by comparing with surgery treatment. METHODS: The clinicopathological data of 320 patients with EGC who were treated in Peking University First Hospital between January 2010 and December 2017 were retrospectively reviewed, in which there were 198 cases of surgical procedure and 122 cases of ESD. Characteristics of lymph node metastasis in EGC were analyzed, and lymph node metastasis of EGC with ESD absolute and expanded indications were summarized. The long-term efficacy of ESD and surgical treatment of EGC were compared to evaluate the rationality of absolute and expanded indications of ESD. RESULTS: Lymph node metastasis was detected in 22 (11.1%) of 198 patients. Univariate analysis showed a positive relationship between tumor size (χ2=5.525, P=0.019), depth of invasion(χ2=8.235, P=0.004), histological type (χ2=6.323, P=0.012), lymphovascular invasion (χ2=12.273, P < 0.001) and lymph node metastasis in EGC. Multivariate analysis revealed that depth of invasion(Wald=7.575, P=0.006) and histological type (Wald=6.317, P=0.012) were independent relative factors of lymph node metastasis in EGC. The lymph node metastasis rates of the patients with absolute and expanded ESD indications were both 0%. The 5-year survival rates of the patients who met ESD absolute indication receiving surgery treatment and ESD were 97.6% and 97.9% respectively, and the difference between the two groups was not statistically significant(χ2=0.014, P=0.907).The 5-year survival rates of the patients who met ESD expanded indication receiving surgery treatment and ESD were 96.5% and 91.7% respectively, the difference between the two groups was not statistically significant(χ2=1.061, P=0.303). CONCLUSION: Lymph node metastasis in EGC is mainly correlated with depth of invasion and histological type. Our data indicate that ESD procedure for EGC is comparable to surgery in terms of long-term efficacy in both absolute and expanded indications. However, some studies of a large sample size are still needed for more confirmation.

[Identification of chemical constituents in Qizhi Tongluo Capsules by UPLC-Q-Orbitrap HRMS].

Qizhi Tongluo Capsules, which is composed of 26 herbal drugs, is mainly used as the assistant therapy for apoplexy sequelae. The chemical composition of Qizhi Tongluo Capsules was complex,but its chemical constituents and the pharmacodynamic material basis remain unreported. The ultrahigh performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry(UPLC-Q-Orbitrap HRMS) was applied to recognize the chemical constituents of Qizhi Tongluo Capsules. The analysis was performed on a Waters Acquity UHPLC HSS T3 column with acetonitrile-water as the mobile phase for gradient elution. Heated electrospray ionization(HESI) in both positive and negative ion modes was adopted to collect the data. The chemical constituents were identified and confirmed by analyzing the accurate molecular weight, the mass fragmentation pattern, and comparing with the mass data from the reference substances and literature. A total of 119 components were identified, including 22 flavones, 12 saponins, 10 salvianolic acids, 5 butylphthalides, 4 anthraquinones, 4 monoterpenoid glycosides, 2 caffeoyl-quinic acids, 2 coumarins, 2 alkaloids, and 1 stilbene, as well as the common constituents in the herb, such as amino acids and organic acids. The chemical constituents of Qizhi Tongluo Capsules were characterized rapidly for the first time in this study, laying a foundation for the further analysis of active compounds and quality control.

Endothelin-1 induces chondrocyte senescence and cartilage damage via endothelin receptor type B in a post-traumatic osteoarthritis mouse model.

OBJECTIVES: This study aimed to investigate the role of endothelin-1 (ET-1), originally known as the potent vasoconstrictor, and its receptors in chondrocyte senescence and osteoarthritis (OA) development. METHOD: Temporal changes of ET-1 and its receptors with OA development were characterized in a posttraumatic OA (PTOA) mouse model at time zero, 1-month and 4-month after surgical induction via destabilization of medial meniscus (DMM). A transgenic ET-1 overexpression (TET-1) mouse model was deployed to assess the impact of upregulated ET-1 on chondrocyte senescence and cartilage degradation. Effects of endothelin receptor blockade on chondrocyte senescence and OA development were further examined both in vitro and in vivo. RESULTS: Local expression of ET-1 in subchondral bone and synovium upregulated after DMM with an increase of plasma ET-1 level from 3.18 ± 0.21 pg/ml at time zero to 6.47 ± 0.34 pg/ml at 4-month post-surgery. Meanwhile, endothelin type B receptor (ETBR) (53.31 ± 2.42% to 83.8 ± 2.65%) and p16INK4a (10.91 ± 1.07% to 28.2 ± 1.0%) positve chondrocytes accumulated in articular cartilage since 1-month prior to cartilage loss at 4-month post-surgery. Overexpressed ET-1 promoted p16INK4a-positive senescent chondrocytes accumulation and cartilage degradation in TET-1 mice. Selective blockade of ETBR, but not ETAR, lowered the expression of p16INK4a in ET-1 or H2O2-induced chondrocyte senescence model, and mitigated the severity of murine PTOA. Intriguingly, reactive oxygen species (ROS) scavenger, Vitamin C, could rescue ET-1-induced chondrocyte senescence in vitro associated with restoration of mitochondrial dynamics. CONCLUSION: ET-1 could induce chondrocytes senescence and cartilage damages via ETBR in PTOA.

[The expression of p53 protein and its clinicopathological features and prognosis of esophageal spindle cell carcinoma].

Objective: To evaluate the association of p53 protein expression with clinicopathological features and prognosis in esophageal spindle cell carcinoma. Methods: A total of 4 439 esophageal squamous cell carcinoma (ESCC) patients who underwent radical esophagectomy without neoadjuvant therapy between May 2010 and May 2019 were included. The HE slides and clinicopathological parameters were reviewed. Among these, there were 63 cases of esophageal spindle cell carcinoma; p53 protein expression was evaluated by immunohistochemistry (IHC) and its correlation with clinicopathological parameters and patients' outcome was analyzed. Results: The 63 esophageal spindle cell carcinoma accounted for 1.4% (63/4 439) of all ESCC. Of the 63 patients there were 55 males and 8 females, male to female ratio was 7∶1. The p53 protein mutation expression rate was 77.8% (49/63), including 14 cases with wild-type expression, 22 with nonsense mutation expression, and 27 with missense mutation expression. The concordance rate of p53 protein expression between carcinoma components and spindle cell components was 100%. Survival analysis showed that p53 protein mutation expression was significantly correlated with overall survival (OS, P=0.044), patients with p53 protein mutation expression had poorer OS. Conclusion: p53 protein expression is highly concordant in the squamous cell carcinoma components and spindle cell components of esophageal spindle cell carcinoma; its mutation expression is associated with poor outcome of the patients.

LncRNA-599554 sponges miR-15a-5p to contribute inductive ability of dermal papilla cells through positive regulation of the expression of Wnt3a in cashmere goat

BACKGROUND: Long non-coding RNAs (lncRNAs), as post-transcriptional regulators, were thought to function in the inductive property of dermal papilla cells (DPCs) in cashmere goat. Previously, lncRNA-599554 was identified in secondary hair follicle (SHF) of cashmere goat, but its functional significance is unknown. RESULTS: In the present investigation, we verified that lncRNA-599554 had significantly higher expression at the anagen dermal papilla of cashmere goat SHF than that at telogen. Based on overexpression and knockdown techniques, we found that lncRNA-599554 contributes the inductive property of DPCs of cashmere goat, which was assessed by detecting the changes in the expression of several typical indictor genes in DPCs including ET-1, SCF, Versican, ALP, Lef1 and Ptc-1. Based on RNA pull-down assay, we verified that lncRNA-599554 directly interacted with chi-miR-15a-5p. Also, we showed that lncRNA-599554 positively regulated the Wnt3a expression in DPCs but which did not appear to involve its modulating of promoter methylation. Based on the use of Dual-luciferase reporter assays, our data indicated that lncRNA-599554 regulated the Wnt3a expression through chi-miR-15a-5p-mediated post-transcriptional level. CONCLUSIONS: We showed that lncRNA-599554 contributes the inductive property of DPCs in cashmere goat which might be achieved through sponging chi-miR-15b-5p to promote the Wnt3a expression. The results from the present investigation provided a novel insight into the functional mechanism of lncRNA-599554 in the SHF regeneration of cashmere goat along with the formation and growth of cashmere fiber.

[Clinical short-term and long-term efficacy of endoscopic submucosal dissection in the treatment of early low rectal cancer and precancerous lesions].

Objective: To evaluate the short-term and long-term efficacy of endoscopic submucosal dissection (ESD) in the treatment of early low rectal cancer and precancerous lesions. Methods: Inclusion criteria: (1) Distance from the lower margin of tumor to the anal was ≤ 5 cm. (2) Early low rectal cancers were any size rectal epithelial tumors with infiltration depth limited to the mucosa and submucosa, which were diagnosed by postoperative pathology as high-grade intraepithelial neoplasia or adenocarcinoma of the rectum with infiltration depth of intramucosal or submucosal cancer (M or SM stage). (3) Precancerous lesions included adenoma and low-grade intraepithelial neoplasia of the rectum. (4) Patients received ESD treatment. Patients with tumor invasion depth over submucosa by pathology were excluded. From January 2008 to January 2018, 63 patients meeting the above criteria in Peking University First Hospital were enrolled in this descriptive cohort study. The disease characteristics, clinical manifestations, pathological types, treatment time, hospitalization time, en bloc resection rate (resection of the whole lesion), complete resection rate (both the horizontal and vertical incision margins were negative), postoperative complications and follow-up results were analyzed. Cummulative survival rate was calculated by Kaplan-Meier. Results: The diameter of the lesion was (29.0±23.4) mm and the distance from the lesion to the anus was (2.7±1.8) cm. The median operation time was 45.0 (range, 10.0 to 360.0) minutes, the median hospitalization time was 3.0 (range, 2.0 to 12.0) days, en bloc resection rate was 100%, complete resection rate was 96.8% (61/63), and 1 case (1.6%) had postoperative bleeding. The follow-up rate was 87.3% (55/63) and the median follow-up time was 57.9 (range, 15.6 to 121.1) months. No local recurrence was found during the follow-up period and the 5-year survival rate was 100%. Conclusion: Short- and long-term efficacy of ESD are quite good in the treatment of patients with early low rectal cancer and precancerous lesions.