AIM: To evaluate the changes of microRNA (miRNA) in hepatocytes during hydrogen peroxide-induced oxidative stress injury, and to observe the alleviating effect of mesenchymal stem cell-conditioned medium (MSC-CM) in this progress.METHODS: The hepatocyte oxidative stress injury model was established using hydrogen peroxide and human normal liver cell line L02.MSC-CM was prepared using centrifugation and filter.The effects of MSC-CM on hepatocyte injury were evaluated by apoptosis analysis, cell viability detection, cell cycle, and mitochondrial membrane po-tential (MMP).Twenty-one differentially expressed miRNAs were selected by gene chip hybridization, in which miR-143, miR-145, miR-301a and let-7a were confirmed by RT-qPCR.Bioinformatics software was utilized to predict target proteins of these miRNAs, and then the proteins were verified by Western blot.RESULTS: MSC-CM markedly attenuated hydrogen peroxide-induced oxidative stress injury by reducing apoptosis, promoting cell viability and regulating cell cycle.The ex-pression of miR-143, miR-145, miR-301a and let-7a, indentified by RT-qPCR, increased under the condition of oxidative stress injury, while decreased after MSC-CM treatment.The expression of miR-143 predicted target proteins, HK2 and ADRB1, decreased under the hydrogen peroxide-exposure, while increased after MSC-CM treatment, which is consistent with the regulatory trend of miR-143.CONCLUSION: MSC-CM might attenuate hydrogen peroxide induced oxidative stress injury via inhibiting apoptosis and regulating some miRNA expression.

In pediatrics, one of the most common diseases is infectious diseases, which is caused by pathogens invading the body and accompanied by local tissue damage or systemic inflammatory reactions.Children′s undeveloped immunity and the consequent rapid progress of the disease urge early diagnosis and timely treatment.However, there is no significant specificity for the clinical symptoms of various infectious diseases, especially in the early stage.Pathogen detection, as a diagnostic gold standard, is not beneficial for the early diagnosis due to the strict requirements for the test environment, long time consumption, low positive rate, and easy contamination.Compared to other methods, blood routine is the most basic and inexpensive test item in clinical practice with the advantage of simple operation and short time.It mainly detects the quality, quantity, and morphological changes in blood components, such as white blood cells, red blood cells, and platelets.It is a valuable test for early diagnosis, differential diagnosis, and evaluation of the severity as well as the treatment efficacy for infectious diseases.Therefore, blood routine test currently is the most preferred method for clinical diagnosis of infectious diseases in pediatrics.

Objective To investigate the correlation of serum concentration of B vitamins B1,B6,B12) with the incidence of epilepsy.Methods We recruited 50 newly diagnosed patients with epilepsy (not receiving antiepileptic drugs) and 50 patients without epilepsy.Serum concentrations of B vitamins (B1,B6,and B12) in both groups were detected with a LK3000V vitamin detector.Results The concentrations of vitamin B1 and B12 in epileptic patients were (132.81 ±7.3l) nmol/L and (221.41 ±26.09) pg/ml,respectively,not significantly different from those in non-epileptic patients [(130.44 ± 5.12) nmol/L,P =0.095 ;(197.87 ± 19.36) pg/ml,P =0.077].The concentration of vitamin B6 in epileptic children was (44.61 ±2.49) μmol/L,significantly lower than that in non-epileptic children [(64.63 ± 5.71) μmol/L,P =0.031].Conclusion Though serum concentrations of vitamin B1 and B12 may not be associated with epilepsy,lower concentration of vitamin B6 in serum may increase the risk of epilepsy.

A 4-year-old boy complained of weakness of the lower limbs for one and a half month.The child had been diagnosed as X-linked agammaglobulinemia (XLA) at 1-year old.In recent one and a half month,he gradually suffered from activity intolerance and fatigue,inability to jump and run,staggering gait and slow speech.All the symptoms above indicated deteriorating motor function.The brain magnetic resonance imaging revealed abnormal signals in white matter and brain atrophy.The cerebrospinal fluid analysis detected the presence of oligoclonal immunoglobulin G band.In short term after intravenous immunoglobulin and methylprednisolone treatment,the boy's lower extremity function and speech speed were slightly improved.However,at 1-year follow-up,the boy's condition became even worse.The child could not sit without support and had difficulty in swallowing.The child could not speak or follow any commands.Neurological examination revealed spastic quadriplegia and pseudobulbar palsy.Progressive neurodegeneration is not a common syndrome in patients with XLA.Brain biopsy is an important approach clinically to find out etiology.

Objective: To explore the effects of O-GlcNAc glycosylation and its key enzyme OGT on the biological behaviors and etoposide (Vp16) -induced apoptosis of Nalm-6 cells. Methods: Low O-GlcNAc modified Nalm-6 cells model was established with Alloxan, an inhibitor of OGT. The influence of Alloxan on Nalm-6 cells proliferation was checked by CCK-8 assay, apoptosis and cell cycle by flow cytometry. Nalm-6 cells were treated with different concentrations of Vp16 for 12 h, and then the O-GlcNAc level and the expressions of OGT were examined by Western blot. After treating Nalm-6 with Alloxan for 24 h and then 5 μg/ml of Vp16 for 12 h, the apoptosis of different groups were measured with flow cytometry, and the expression of apoptosis-associated proteins Bax and Bcl-2 were examined by Western blot. Results: With the concentration of Vp16 increasing, the O-GlcNAc modified levels of total protein and the expression of OGT were up regulated (P<0.05, n=6) ; Alloxan could slow down the proliferation capacity, induce apoptosis[ (15.190±2.539) % vs (21.910±4.105) %, P=0.007], arrest cell cycle[G1 phase: (43.534±4.453) % vs (57.322±6.091) %, P=0.003; S phase: (50.747±5.937) % vs (37.201±4.661) %, P=0.001]. Alloxan could inhibit the apoptosis caused by Vp16[ (75.195±13.845) % vs (52.741±10.815) %, P=0.011]along with Bax decreasing (5.496±1.998 vs 2.950±0.703, P=0.015) and Bcl-2 increasing (0.454±0.125 vs 0.803±0.223, P=0.013) . Conclusion Changes of O-GlcNAc modified level of Nalm-6 cells along with the inhibition of OGT could influence the biological behaviors and inhibit apoptosis induced by Vp16.

Background and Objectives: Bronchopulmonary dysplasia (BPD) has major effects in premature infants. Although previous literature has indicated that mesenchymal stem cells (MSCs) can alleviate lung pathology in BPD newborns and improve the survival rate, few research have been done investigating significantly differentially expressed genes in the lungs before and after MSCs therapy. The aim of this study is to identify differentially expressed genes in lung tissues before and after hAD-MSC treatment. Methods: and Results: Human amnion-derived MSCs (hAD-MSCs) were cultured and met the MSCs criteria for cell phenotype and multidirectional differentiation. Then we confirmed the size of hAD-MSCs-EXOs and their expressed markers. An intratracheal drip of living cells showed the strongest effect on NHLI compared to cellular secretions or exosomes, both in terms of ameliorating pulmonary edema and reducing inflammatory cell infiltration. Through gene chip hybridization, PCR, and western blotting, acylaminoacyl-peptide hydrolase (APEH) expression was found to be significantly decreased under hyperoxia, and significantly increased after hAD-MSC treatment. Conclusions The intratracheal transplantation of hAD-MSCs ameliorated NHLI in neonatal rats through APEH.

OBJECTIVETo evaluate the feasibility and characteristics of human engraftment in HLA disparate cord blood transplantation.

METHODSTwo human HLA-haploidentical or HLA-mismatched cord blood units were transplanted into sublethally irradiated severe combined immunodeficiency (SCID) mice. The characteristics of engraftment, hematopoietic and immunological reconstitution between the two groups were compared.

RESULTSTwo mixed cord blood units can engraft in SCID mice with donor-recipient chimerism and reconstitute hematopoiesis and immunological functions. No unfavorable factors had been observed. Only one of the two cord blood units which had higher colony forming ability in vitro could engraft in most SCID mice as shown by HLA-DQB(1) gene detection. Two HLA-haploidentical cord blood units were simultaneously engrafted in 3 SCID mice.

CONCLUSIONDouble HLA-haploidentical or HLA-mismatched cord blood can engraft in SCID mice and reconstitute hematopoietic and immunological functions. HLA disparity has no significant effect on survival and engrafting rate. However, in less HLA disparity group, two cord blood units were prone to engraft simultaneously.

Animals ; Antigens, CD ; immunology ; Cord Blood Stem Cell Transplantation ; methods ; Disease Models, Animal ; Female ; Fetal Blood ; immunology ; metabolism ; Flow Cytometry ; HLA Antigens ; genetics ; immunology ; Hematopoiesis ; Humans ; Mice ; Mice, SCID ; Random Allocation ; Severe Combined Immunodeficiency ; immunology ; physiopathology ; surgery ; Survival Analysis ; Transplantation, Heterologous

Objective To study perioperative period conditions of total and partial colpocleisis with severe pelvic organ prolapse(POP)in elderly patients. Methods From Jan. 2014 to Oct. 2017286 severe POP elderly patients underwent total or partial colpocleisis. The mean age was(76.0±5.1)years(60-90 years). Of which,220 patients(76.9％, 220/286)presented more than one kind of medical disease. The operative time, intraoperative blood loss, hospital stay after surgery, postoperative complications, preoperative urinary dysuria symptoms of total and partial colpocleisis in treatment of 286 patients were analyzed. Results In 286 patients, 172 patients(60.1％)underwent total colpocleisis and 114 patients(39.9％) underwent partial colpocleisis. Totally 48 patients(16.8％)in 286 patients underwent anti-urinary incontinence procedure. The mean operating time of 286 patients was(67 ± 22)minutes, the mean blood loss was(52 ±57)ml, the mean hospital stay after surgery was(5.9 ± 1.9)days(2-16 days). The rate of postoperative complications was 8.7％(25/286). Those complications including 1 case of bladder injury during operation, 3 cases of postoperative pelvic hematoma, 1 case of intestinal obstruction, 5 cases of atrial fibrillation, 10 cases of postoperative morbidity, 3 cases of pulmonary embolism, 1 case of lower limb thrombosis, 1 case of urinary retention underwent electrical stimulation treatment because of ineffective application of urinary retention catheter. The mean operating time was(67±22)minutes in total colpocleisis and(68±20) minutes in partial colpocleisis(P>0.05), the mean blood loss was(58±62)ml in total colpocleisis and(41± 45)ml in partial colpocleisis(P>0.05), the mean hospital stay after surgery was(5.9 ± 1.9)days in total colpocleisis and(6.0 ± 1.8)days in partial colpocleisis(P>0.05), the rate of post operative complications was 8.1％(14/172) in total colpocleisis and 9.6％(11/114)in partial colpocleisis(P>0.05), respectively;those four of comparisons showed no significant difference. There were 129 patients(45.1％, 129/286)with voiding difficulty before surgery;the mean postvoid residual volumes of the above two operation types of people after operation were(35 ± 43)and(34 ± 41)ml, which showed no significant difference(P>0.05). Conclusions Colpocleisis is a safe and effective management in selected elderly patients with severe POP, who no longer desire to maintain vaginal coital function. There is no significant difference in the perioperative period conditions between total and partial colpocleisis. The choice of surgical procedure is based on factors such as the presence or absence of malignancy in the uterus, age and willingness of the patient, and general condition.

Objective:To compare the clinical value of early and deferred empirical antifungal strategies in febrile neutropenic children with acute leukemia.Methods:A total of 101 cases of febrile neutropenic children with acute leukemia hospitalized in Qilu Hospital of Shandong University from January 2019 to June 2021 were divided into two groups according to different empirical antifungal strategies.There were 41 cases in early group in which antifungal therapy was given within 4 days of fever, and 60 cases in deferred group in which antifungal therapy was not given within 4 days of fever.Outcomes such as time to stable defervescence, positive diagnosis rate of invasive fungal disease, incidence of severe pneumonia, rate of transference to PICU, exposure time and costs of antifungal agents, and infection-related hospitalization days were compared between two groups.Results:There were no significant differences in time to stable defervescence[5 (4, 7) days vs.5 (3, 7) days, P=0.986], positive diagnosis rate of invasive fungal disease[9.8%(4/41) vs.8.3%(5/60), P=1.000], incidence of severe pneumonia[19.5%(8/41) vs.10.0%(6/60), P=0.174], and rate of transference to PICU[2.4%(1/41)vs.0(0/60), P=0.406] between two groups.Exposure time of antifungal agents was longer in early group than that in deferred group[10 (6, 12)days vs.0 (0, 6)days, P<0.001]. Costs of antifungal agents were higher in early group than those in deferred group[0.78(0.51, 0.95)ten thousand yuan vs.0(0, 0.44)ten thousand yuan, P<0.001]. Infection-related hospitalization days were longer in early group than those in deferred group[16 (10, 21) days vs.9(6, 13)days, P<0.001]. Conclusion:For febrile neutropenic children with acute leukemia, clinical effect of early empirical antifungal strategy is not superior to that of deferred empirical antifungal strategy.Pediatricians should make reasonable antifungal decisions according to overall situation of patients.

OBJECTIVETo investigate the role of CCL20/CCR6/Th17 axis in vascular invasion and metastasis of primary hepatocellular carcinoma (HCC).

METHODSExpression levels of CCL20 mRNA in the normal human liver cell line L-02, and human hepatocellular carcinoma cell lines Hep3B, Huh7 and HepG2 were quantified by using SYBR green real time PCR. CCL20 secretions from these cell lines were quantified by using ELISA. The chemotactic effect of HCC cell line Hep3B on human peripheral blood mononuclear cells was determined by using transwell chemotaxis assay. Pre-therapy serum levels of IL-1α, IL-1β, IL-6, IL-8, IL-10, IL-17, IL-23, IFN-γ, TNF-α and CCL20 in 93 patients with HCC were measured by using 9-plex array and ELISA. All the patients were chronic hepatitis B virus associated HCC, and 51 cases were those with vascular invasion and metastasis (metastasis group) and 42 cases were not (non-metastasis group). CCL20 and CCR6 mRNA expressions in the HCC and tumor-adjacent tissues were determined by using SYBR Green real time PCR in 41 patients, among them, 20 cases were from the group of patients with metastasis and 21 cases were from the group of patients without metastasis. The CCL20 expression was further determined by immunohistochemistry.

RESULTSThe HCC cell lines expressed and secreted higher amount of CCL20, which effectively recruited CCR6(+) T cells. Pre-therapy serum levels of CCL20 in 93 HCC patients were (38.2 ± 28.4)pg/ml, significantly increased than those with benign hepatic hemangiomas [(7.8 ± 17.8)pg/ml, P < 0.01]. In addition, the serum levels of CCL20 were positively correlated with the tumor diameters in HCC patients (r = 0.32, P = 0.0018). CCL20 was dominantly expressed in the cytoplasm in HCC cells, and it was also expressed by some infiltrating immune cells. The mRNA expression levels of CCL20 of the tumor tissues were significantly higher than that in the tumor-adjacent tissues (P < 0.05). Multivariate logistic regression analysis showed that serum levels of IL-17 and CCL20 were independent risk factors of metastasis in HCC patients (P < 0.05 for both). CCL20 mRNA showed no statistically significant differences between patients with metastasis and without metastasis in both tumor tissues and tumor-adjacent tissues (P > 0.05 for both). But the patients with metastasis showed significantly higher expressions of CCR6 both in their tumor [5.75 (1.79, 19.13)]and tumor-adjacent tissues [7.99 (4.49, 19.54)] than those with non-metastasis [1.69 (0.76, 2.87) and 3.58 (1.84, 4.32), P < 0.05 for both].

CONCLUSIONCCL20/CCR6/Th17 axis may promote vascular invasion and metastasis hepatocellular carcinoma.

Bile Duct Neoplasms ; Carcinoma, Hepatocellular ; metabolism ; Chemokine CCL20 ; metabolism ; Humans ; Interleukin-10 ; metabolism ; Interleukin-17 ; metabolism ; Interleukin-23 ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Leukocytes, Mononuclear ; Liver Neoplasms ; metabolism ; RNA, Messenger ; Th17 Cells ; Tumor Necrosis Factor-alpha ; metabolism