Objective To explore the effect of melatonin ( MLT) on the initiation of puberty in female mice and on the expression level of phosphatidylinositol-3-kinases ( PI3K)/protein kinase B ( Akt)/mammalian target of rapamycin (mTOR) signaling pathway in the frypothalamus. Methods Seventy-eight 20-day-old female KM mice were randomly divided into melatonin (MLT) group and normal saline (NS) group, with 39 mice in each group. Starting at 22 days of age, the MLT group was given a subcutaneous injection of 1 mg/kg melatonin and the NS group was given an equal volume of saline. Thirty-two days of age were selected as the sampling point before puberty initiation and 13 mice were executed in each of the two groups, while 37 and 42 days of age were selected as the sampling point after puberty initiation and 13 mice were executed in each of the two groups. Observation of vaginal opening time in mice, weighing of ovaries and uterus to calculate organ indices. HE staining to observe the number of ovarian corpora lutea. The levels of serum luteinizing hormone (LH)were determined by ELISA. The mRNA and protein expression levels of PI3K/Akt/mTOR pathway in frypothalamus were detected by Real-time PCR and Western blotting. Results Compared with the normal saline group, mice in the melatonin group had significantly delayed vaginal opening time ( P < 0. 05 ) , decreased significantly ovarian and uterine volume and index (P<0. 05) , decreased significantly serum LH levels (P<0. 05) , and decreased significantly mRNA and protein expression levels of the frypothalamic PI3K/Akt/mTOR pathway (P<0. 05). Conclusion Melatonin delays puberty initiation in mice by a mechanism that ma)' be related to inhibition of the hypothalamic PI3K/Akt/mTOR signalling pathway.

Objective: To investigate the levels of Th9 cells and interleukin-9 (IL-9), and to assess the regulatory activity of Th9/IL-9 to anti-tumor immune response in patients with gastric cancer. Methods: Thirty-four patients with gastric cancer who received operation in the First Affiliated Hospital of Xinxiang Medical University between October 2018 and August 2019 were included. Twenty individuals who received physical examination in the same period were also enrolled. Peripheral blood was collected, and then plasma and peripheral blood mononuclear cells (PBMCs) were isolated. Tumor-infiltrating lymphocytes (TILs) and autologous gastric cancer cells were isolated from resected gastric cancer tissues. CD4(+) T cells, CD8(+) T cells, and CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells were purified from PBMCs and TILs. Plasma IL-9 level was measured by enzyme linked immunosorbent assay (ELISA). The percentage of CD3(+) CD4(+) IL-9(+) Th9 cells in PBMCs and TILSs was assessed by flow cytometry. The mRNA levels of IL-9 and transcriptional factors purine-rich nucleic acid binding protein 1 (PU.1) were semi-quantified by real-time quantitative polymerase chain reaction (RT-qPCR). PBMCs and TILs from gastric cancer patients were stimulated with recombinant human IL-9. Cellular proliferation was measured by cell counting kit-8. The phosphorylation levels of signal transducer and activator of transcription 3 (STAT3) and STAT6 were investigated by western blot. Cytokine production was measured by ELISA. Purified CD8(+) T cells from TILs of gastric cancer patients were stimulated with recombinant human IL-9. CD8(+) T cells and autologous gastric cancer cells were cocultured in direct contact and indirect contact manner. The percentage of target cell death was calculated by measuring the lactate dehydrogenase (LDH) level. These cretion of γ-Interferon (γ-IFN) and tumor necrosis factor-α (TNF-α) was measured by ELISA. CD4(+) CCR4(-)CCR6(-)CXCR3(-)cells, CD8(+) T cells, and autologous gastric cancer cells were directly cocultured, and anti-IL-9 neutralizing antibody was added. The target cell death was measured. Results: The percentages of CD3(+) CD4(+) IL-9(+) Th9 cells in PBMCs of control group and PBMCs of gastric cancer group were (1.21±0.25)% and (1.14±0.19)%, respectively. The difference was not statistically significant (P=0.280). The percentage of CD3(+) CD4(+) IL-9(+) Th9 cells in TILs of gastric cancer group was (2.30±0.55)%, which was higher than those in PBMCs of control group and PBMCs of gastric cancer group (P<0.001). The plasma IL-9 level in control group and gastric cancer group were (5.04±1.51) and (4.93±1.25) ng/ml. The difference was not statistically significant (P=0.787). The relative levels of IL-9 mRNA in PBMCs of control group and PBMCs of gastric cancer group were 1.33±0.39 and 1.36±0.27. The difference was not statistically significant (P=0.691). The relative level of IL-9 mRNA in TILs of gastric cancer group was 2.90±0.75, which was higher than those in PBMCs of control group (P<0.001) and PBMCs of gastric cancer group (P<0.001). The relative levels of PU.1 mRNA in PBMCs of control group and PBMCs of gastric cancer group were 1.21±0.12 and 1.20±0.11. The difference was not statistically significant (t=0.21, P=0.833). PU.1 mRNA relative level in TILs of gastric cancer group was 2.81±0.65, which was higher than those in PBMCs of control group (P<0.001) and PBMCs of gastric cancer group (P<0.001). Recombinant human IL-9 stimulation did not affect the proliferation of PBMCs and TILs of gastric cancer patients (P>0.05), but elevated the phosphorylation level of STAT6 and induced the secretions of γ-IFN, IL-17, and IL-22 by TILs (P<0.05). In direct contact culture system, IL-9 stimulation promoted tumor-infiltrating CD8(+) T cells-induced autologous gastric cancer cell death [(20.62±2.27)% vs. (16.08±2.61)%, P<0.01)]. In indirect contact culture system, IL-9 stimulation did not increase CD8(+) T cell-induced autologous gastric cancer cell death [(5.21±0.70)% vs. (5.31±1.22)%, P=0.998)]. However, the secretion levels of γ-IFN were elevated in response to IL-9 stimulation in both culture systems [direct contact culture system: (100.40±12.05) pg/ml vs. (76.45±8.56) pg/ml; indirect contact culture system: (78.00±9.98) pg/ml vs. (42.09±10.71) pg/ml; P<0.01]. The TNF-α secretion level did not significantly changed (P>0.05). In direct contact culture system, the percentage of target cells was (22.01±3.05) % and γ-IFN secretion level was (104.5±12.84) pg/ml in CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells+ CD8(+) T cells+ gastric cancer cells group, which was higher than (16.08±2.61)% and (76.45±8.56) pg/ml in CD8(+) T cells+ gastric cancer cells group (P<0.01). However, the percentage of target cells was (14.47±3.14)% and γ-IFN secretion level was (70.45±19.43) pg/ml in CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells+ CD8(+) T cells+ gastric cancer cells+ anti-IL-9 neutralizing antibody group, which were lower than those in CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells+ CD8(+) T cells+ gastric cancer cells group (P<0.01). Conclusion: Tumor-infiltrating Th9 cells and the secreting IL-9 promote the activity of CD8(+) T cells in gastric cancer patients, and enhance anti-tumor immune response.
Humans ; CD8-Positive T-Lymphocytes ; Stomach Neoplasms/pathology* ; Tumor Necrosis Factor-alpha/metabolism* ; Lymphocytes, Tumor-Infiltrating/pathology* ; Interferon-gamma/metabolism* ; RNA, Messenger/metabolism* ; Antibodies, Neutralizing/metabolism*

Kidney is an essential organ in human body with multiple physiological functions. However, there is 10 % population worldwide with renal disease. It is urgent to generate a model which is more similar with kidney at structural and functional level to study renal disease. The rise of in vitro differentiation technology from pluripotent stem cells gives regeneration medicine and precise medicine new energy. This study mimics kidney development in vitro by inducing human pluripotent stem cells including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) into kidney progenitor cells, and further forming nephrons, which is the structure and function unit in kidney. Human pluripotent stem cells were differentiated into primitive streak through activating WNT pathway while inhibiting TGF-(B signaling. Afterward, the primitive streak spontaneously differentiated into intermediate mesoderm. Then, we induced intermediate mesoderm cells into kidney progenitor cells through FGF pathway. The FACS analysis data indicated kidney progenitor cells were up to 51. 5%-61. 9% in total cell population. Immuno-stai-ning results showed these structures contained podocytes of glomerulus, proximal tubule, and distal tubule. This kidney differentiation protocol is stable, high-efficient, and well repeatable. This research provides a novel platform for early human kidney development study, kidney disease modeling, and drug screening.

【Objective】 To explore the influence of anti-HLA-Ⅰ with different mean fluorescence intensity (MFI) on the efficacy of HLA-A and -B gene matching platelet transfusion, so as to provide scientific data for clinical platelet gene matching transfusion strategy. 【Methods】 A total of 81 PTR patients had applied for HLA-Ⅰgene matched platelets from the platelet gene database established by our laboratory, and 28 (MFI <5 000) of them needed further avoiding of partial donor-specific antibodies and they were enrolled as the research subjects. According to the platelet MFI value of HLA-Ⅰ antibody-targeting antigen, they were divided into negative transfusion group (MFI <500) (group A) and positive transfusion groups (MFI≥500) ; the latter were further divided into group B (500≤MFI <1 000), group C (1 000≤MFI <3 000) and group D (MFI≥3 000) according to MFI value. Corrected count increment (CCI) in platelet count was used to compare the platelet transfusion effect in 4 groups. 【Results】 Among 28 platelet recipients with MFI <5 000, 19(67.86%) patients successfully received 72 effective transfusions. The first CCI (×10⁹/L) in groups A, B, C and D were 10.27±7.46, 7.58±4.75 (P>0.05), 17.36±7.63 (P>0.05) and -0.77±2.30 (P<0.05), respectively. There was no statistical difference among group A, B and C. 【Conclusion】 The application of HLA-Ⅰ gene matching platelets in PTR patients can adjust the MFI threshold(<2 000) appropriately according to the patient′s condition without compromising the platelet transfusion effect.

OBJECTIVE: To observe the clinical effect of high suspension and low incision (HSLI) surgery on mixed haemorrhoids, compared with Milligan-Morgan haemorrhoidectomy. METHODS: A multi-centre, randomized, single-blind, non-inferiority clinical trial was performed. Participants with mixed haemorrhoids from Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing Rectum Hospital, Air Force Medical Center of People's Liberation Army of China, and Puyang Hospital of Traditional Chinese Medicine were enrolled from September 2016 to March 2018. By using a blocked randomization scheme, participants were assigned to two groups. The experimental group was treated with HSLI, while the control group was treated with Milligan-Morgan haemorrhoidectomy. The primary outcome was the clinical effect evaluated at 12 weeks after operation. The secondary outcomes included the number of haemorrhoids treated during the operation, pain scores, use of analgesics, postoperative oedema, wound healing, incidence of anal stenosis, anorectal manometry after operation, as well as surgical duration, length of stay and total hospitalization expenses. A safety evaluation was also conducted. RESULTS: In total, 246 eligible participants were enrolled, with 123 cases in each group. There was no significant difference in the clinical effect between the two groups (100.00% vs. 99.19%, P>0.05). Compared with the control group, the number of external haemorrhoids treated during the operation and the pain scores after operation were significantly reduced in the experimental group (P<0.05 or P<0.01); the patient number with wound healing at 2 weeks after operation and the functional length of anal canal at 12 weeks after operation were significantly increased in the experimental group (P<0.05). There was no significant difference in the incidence of anal stenosis, the numbers of patients using analgesics and patients with postoperative oedema between the two groups after operation (P>0.05). The surgical duration and length of stay in the experimental group were significantly longer than those in the control group, and the total hospitalization expense was significantly higher than that in the control group (all P<0.05). No adverse events were reported in either group during the whole trial or follow-up period. CONCLUSION HSLI had the advantages of preserving the skin of anal canal completely, alleviating postsurgical pain and promoting rapid recovery after operation. (Registration No. ChiCTR1900022883).

Malocclusion is one of the three most common oral diseases reported by World Health Organization(WHO). In China, its incidence rate is rising. Malocclusion seriously affects the dental and maxillofacial function, facial appearance and growth development of nearly 260 million children in China, and what is more, it affects their physical and mental health development. Malocclusion occurrence is related to genetic and environmental factors. Early treatment of malocclusion can create a good dental and maxillofacial development environment, correct abnormal growth and control the adverse effects of abnormal genetic factors. It can effectively reduce the prevalence of children's malocclusion and enhance their physical and mental health. This is an urgent need from the economic perspective of our society, so it has great practical and social significance. Experts from the project group "standard diagnose and treatment protocols for early orthodontic intervention of malocclusions of children" which initiated by China National Health Institute of Hospital Administration wrote the "China Experts' Consensus on Preventive and Interceptive Orthodontic Treatments of Malocclusions of Children", which aims to guide and popularize the clinical practice, improve the clinical theory and practice level, and accelerate the disciplinary development of early treatment of children's malocclusion in China. The consensus elaborates the harmfulness of malocclusion and the necessity of early treatment, and brings up the principles and fundamental contents. Based on the law of dental and maxillofacial development, this paper puts forward the guiding suggestions of preventive and interceptive treatments in different stages of dental development ranging from fetus to early permanent dentition. It is a systematic project to promote and standardize the early treatment of malocclusion. Through scientific and comprehensive stratified clinical practice and professional training, the clinical system of early treatment of malocclusion in China will eventually be perfected, so as to comprehensively care for children's dental and maxillofacial health, and improve their oral and physical health in China.
Child ; China/epidemiology* ; Consensus ; Dental Care ; Humans ; Malocclusion/prevention & control* ; Orthodontics, Interceptive

We explored the mechanism of patchouli oil in the treatment of inflammatory bowel disease (IBD) based on network pharmacology and differentially expressed genes in macrophages. The chemical composition of patchouli oil was detected by GC-MS, targets for active components were collected through TCMSP and Swiss Target Prediction platform, and targets for treatment of IBD were retrieved from DrugBank, GeneCards, OMIM, PharmGkb, and TTD databases. The intersection targets were merged, Cytoscape software was used to construct the "component-to-intersection target" network, and protein-protein interaction (PPI) network was drawn with String platform. The intersection targets were enriched for GO and KEGG enrichment analysis on Metascape platform, and the molecular docking of AutoDock Vina was used to verify the analysis results. The macrophage chip data was downloaded, and the differential genes were obtained by using R software. KEGG signaling pathway analysis of differentially expressed genes were performed by DAVID platform. Real-time fluorescence quantitative PCR was used to verify the screened components in the cell model in vitro. The 14 main components of patchouli oil corresponded to 112 targets, and the intersection obtained 97 common targets of patchouli oil for IBD treatment. GO enrichment analysis yielded 53 items. Eighteen items were obtained by KEGG enrichment analysis, involving cAMP signaling pathway, Notch signaling pathway, adhesion connection, Th17 cell differentiation and other signaling pathways. Molecular docking showed that the selected active components of patchouli oil had good binding activity with the targets. Differentially expressed genes were enriched in inflammatory pathways such as Toll-like receptors, JAK-STAT and NF-κB signaling pathways. q-PCR showed that patchouli oil, patchouli alcohol, pogostone can reduce the mRNA levels of cytokines (TNF-α, IL-1β, IL-6, and IL-23) and up-regulate the mRNA levels of tight junction proteins (occludin and claudin-1) in the inflammatory model of NCM460 normal colon epithelial cells. Patchouli alcohol can significantly reduce the levels of TNF-α, IL-6, and IL-1β inflammatory factors in RAW264.7 macrophages induced by LPS. This study revealed the multi-component, multi-target and multi-pathway of patchouli oil, and confirms the anti-inflammatory effect of patchouli oil and its main components in the inflammatory cell model in vitro and the protection of intestinal epithelial barrier integrity function, which provides a theoretical basis for further elucidating the mechanism of patchouli oil in the treatment of IBD.

OBJECTIVE: To study the clinical features of coronavirus disease 2019 (COVID-19) in children aged <18 years. METHODS: A retrospective analysis was performed from the medical data of 23 children, aged from 3 months to 17 years and 8 months, who were diagnosed with COVID-19 in Jiangxi, China from January 21 to February 29, 2020. RESULTS: Of the 23 children with COVID-19, 17 had family aggregation. Three children (13%) had asymptomatic infection, 6 (26%) had mild type, and 14 (61%) had common type. Among these 23 children, 16 (70%) had fever, 11 (48%) had cough, 8 (35%) had fever and cough, and 8 (35%) had wet rales in the lungs. The period from disease onset or the first nucleic acid-positive detection of SARS-CoV-2 to the virus nucleic acid negative conversion was 6-24 days (median 12 days). Of the 23 children, 3 had a reduction in total leukocyte count, 2 had a reduction in lymphocytes, 2 had an increase in C-reactive protein, and 2 had an increase in D-dimer. Abnormal pulmonary CT findings were observed in 12 children, among whom 9 had patchy ground-glass opacities in both lungs. All 23 children received antiviral therapy and were recovered. CONCLUSIONS COVID-19 in children aged <18 years often occurs with family aggregation, with no specific clinical manifestation and laboratory examination results. Most of these children have mild symptoms and a good prognosis. Epidemiological history is of particular importance in the diagnosis of COVID-19 in children aged <18 years.
Adolescent ; Betacoronavirus ; Child ; Child, Preschool ; China ; Coronavirus Infections ; Humans ; Infant ; Pandemics ; Pneumonia, Viral ; Retrospective Studies

OBJECTIVETo analyze the infection characteristics of sputum and venous blood pathogen in the patients with hematological malignancies and respiratory symptom in the Hematology Department in tropical region and to investigate its drug-resistance so as to provide etiological evidence for clinical treatment.

METHODSThe pathogens and their drug-resistance of 2466 samples in the patients with hematological malignancies and respiratory symptom in the Department were analyzed retrospectively from January 2013 to November 2017. The samples were collected from sputum and venous blood.

RESULTSThe sputum sample culture in patients with hematologic diseases showed that 224 strains were isolated, out of them 98 strains (43.75%) were fungi mainly candida albicans (41 strains); and then 88 Gram-negative strains (39.28%), among them the main pathogenic bacteria were Escherichia coli(22 strains) and klebsiella Klebsiella pneumoniae(12 strains); and then 38 Gram-positive strains (16.96%), among them the main pathogeni-bacteria were Enteroccocus (14 strains) and Gram-positive bacilli (14 strains). The blood samples culture of patients with hematologic diseases showed that 61 strains were isolated, out of them the isolated rate of Gram-positive bactetia was higherst, which accounted for 55.74%(34/61), mainly including staphylococcus lominis (19 strains); and the isolated rate of Gram-negative bacteria was 44.26% (27/61), among them main pathogenic bacteria was Klebsiella pneumoniae (12 strains). The resistance test of pathogenic bacteria to drugs showed that the resistant rate of Gram-negative bacteria to tigecycline, imipenem and atl-962 duenner was lowest, while the Gram-positive pathogenic bacteria such as Enterococcus, Gram-positive bacilli and Staphylococcus aureus were sensitive to vancomycin, tigecycline and linezolid was high.

CONCLUSIONthe patients in hematology department are infected easily in the hospital in tropical region. The main pathogens are fungal strains in the respiratory tract of patients with hematological malignancy according to the sputum culture results. The clinician in tropical regions should choose suitable antibiotics for anti-infective therapy, which is different from the situation in North China or other northern areas.

Bacteria ; Bacterial Infections ; Candida ; Candidiasis ; China ; Drug Resistance, Bacterial ; Hematologic Neoplasms ; Humans ; Microbial Sensitivity Tests ; Retrospective Studies