OBJECTIVETo investigate the effect of ADAM10 inhibitor GI254023X on the proliferation and apoptosis of multiple myeloma H929 cell line and its mechanisms.

METHODSH929 cells were treated with different concentrations of GI254023X, the proliferation-inhibitive curve was assayed and plotted by CCK-8 method, the cell viability and apoptosis were detected by flow cytometry with Annexin V/7-AAD double staining. The cleavage of Notch1 protein (cleaved notch1) was determined by Western blot. The transcripts of Notch1 target gene Hes-1 were detected by real-time PCR.

RESULTSThe GI254023X inhibited the proliferation of H929 cells in the time- and dose- dependent manners. As compared with the control group, the apoptosis of cells increased along with enhancement of GI254023X concentration; The expression of cleaved Notch1 was down-regulated after the treatment with GI254023X. The levels of Hes-1 mRNA transcripts in H929 cells was reduced in GI254023X treated group.

CONCLUSIONGI254023X can remarkably inhibit the proliferation and induce the apoptosis of H929 cells. Its mechanism may be associated with inbihition of Notch1 activation.

ADAM Proteins ; ADAM10 Protein ; Amyloid Precursor Protein Secretases ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Dipeptides ; Down-Regulation ; Humans ; Hydroxamic Acids ; Membrane Proteins ; Multiple Myeloma ; Receptor, Notch1

Objective : To investigate the effects of Lin28 overexpression on the proliferation and osteogenic differentiation of human dental pulp stem cells (hDPSCs) through mTOR signaling pathway. Methods : After transfecting lentiviral vectors of Lin28 gene in hDPSCs , the relative expression of Lin28 was detected by Real⁃time PCR. CCK⁃8 assay was applied to detect the effect on cell proliferation. qRT⁃PCR was used to research the expression levels of alkaline phosphatase (ALP) , osteopontin (OPN) and osteocalcin (OCN) . Western blot assay was processed to investigate the effects on the relative expression levels of ALP and OPN proteins. Alizarin red staining was utilized to detect the mineralized nodules. Results : Compared with the control group , the cell proliferation of transfection group was promoted (P < 0. 05) ; The mRNA and protein expression levels of ALP , OPN and OCN in transfection group were significantly lower than those in control group (P < 0. 05) , the expression level of ALP apparently decreased after the addition of mTOR inhibitor rapamycin (P < 0. 05) ; Alizarin red staining showed that the size and number of mineralized nodules formed in transfection group were markedly declined compared with empty carrier group (P < 0. 05) . Conclusion Overexpression of Lin28 can inhibit the osteogenic differentiation of hDP⁃ SCs through suppress mTOR signaling pathway.

AIM: To describe the clinical features of congenital isolated inferior rectus(IR)palsy with compensatory head posture of facial rotation and to analyze the effect of surgical treatment.

METHODS: Fifteen patients with IR muscle palsy further presenting with abnormal compensatory head posture of facial rotation were retrospectively identified from May 2014 to July 2018. Changes in the degree of facial rotation in patients were evaluated using an orthopedic goniasmometer. Variations in vertical and horizontal deviations were measured through prism and alternate cover tests, and changes in cyclotropia of pre- and postoperation were assessed by objective evaluation of eye torsion with fundus photography. IR resection was the main surgical treatment.

RESULTS: Among the 15 patients, 13 who were subjected to IR muscle resection or IR muscle resection combined with superior rectus muscle recession showed good results after surgery. These results included improved underaction of IR, coordination of eye movement, disappearance of abnormal facial rotation of compensatory head position, and total correction of vertical and horizontal deviation and cyclodeviation. Furthermore, each 1 mm resection of IR can correct 1.54±0.93° incyclotropia. In the other two patients with small vertical deviation who were done by inferior oblique myectomy, the improvement in facial rotation was unsatisfied, and eye motion remained uncoordinated.

CONCLUSION: Congenital isolated IR muscle palsy characterized by compensatory head posture of facial rotation is the main compensatory head posture. The resection of IR is effective and perfect. Mild overcorrection and undercorrection do not influence curative effect.

Infant, Newborn ; Humans ; Macrophage Activation Syndrome/diagnosis*

【Objective】 To investigate the feasibility of prostatic exosomal protein (PSEP) detection kit in the diagnosis of histological prostatitis (HP) in patients with benign prostatic hyperplasia (BPH), and to explore the correlation between PSEP and other clinical parameters. 【Methods】 A total of 104 patients with BPH or BPH plus HP treated during Nov.2021 and Nov.2022 were involved. The patients were instructed to fill out the International Prostate Symptom Score (IPSS) scale independently before surgery. Clinical data such as prostate volume, residual urine volume, free prostate specific antigen (fPSA), total prostate specific antigen (tPSA), and fPSA/tPSA were collected. Preoperative midstream morning urine was collected for PSEP detection. 【Results】 The sensitivity and specificity of PSEP in the diagnosis of BPH were 93.51% and 70.37%, respectively, which were highly consistent with the postoperative pathological diagnosis results (Kappa=0.663). Serum PSEP level was positively correlated with tPSA level (r=0.242, P=0.040). 【Conclusion】 PSEP has a high clinical diagnostic value in the diagnosis of HP, which can provide a reliable basis for the diagnosis of HP in BPH patients and improve the diagnosis rate.

Objective:To observe the effect of braces combined with insoles on scoliosis and walking performance in patients with adolescent idiopathic scoliosis (AIS). Methods:From September, 2019 to September, 2020, 42 subjects with AIS were selected and randomly divided into brace group (n = 21) and brace combined with insole group (n = 21). Both groups received two-month routine rehabilitation, including braces for 22 to 23 hours a day and gymnastics for 30 minutes a day. The brace combined with insole group additionally wore insoles, at least eight hours a day for two months. Meanwhile, 32 even-aged adolescents were recruited as healthy controls. Firstly, gait and plantar pressure of 42 patients and 32 healthy adolescents were compared to find out abnormal indicators. Secondly, the scoliosis and above abnormal indicators were compared between the brace group and the brace combined with insole group Results:The center of pressure excursion index (CPEI) was higher in AIS group than in the healthy group (F = 3.120, P < 0.05), and there was no significant difference in walking speed, gait cycle and phase between two groups (P>0.05). An obvious imbalanced pressure was observed between the medial and lateral heel of the single foot and the bilateral foot in AIS patients (P < 0.05). After treatment, the Cobb's angle decreased in AIS patients (t > 7.552, P < 0.001), however, no difference was found between the brace group and the brace combined with insole group (t = 0.459, P > 0.05); the CPEI decreased (t = 2.209, P < 0.05), and the pressure in medial and lateral heel as well as the left and right foot tended to be balanced (t > 2.306, P < 0.05) in the brace combined with insole group, and were better than that of the brace group (|t| > 2.319, P < 0.05). Conclusion:Plantar pressure distribution shows obvious local and global asymmetric changes in AIS patients. The efficacy of insoles on the scoliosis is limited, but the insole can effectively improve the abnormal biomechanics and balance the force.

OBJECTIVETo assess the effects of warm ischaemia time (WIT) on renalfunction after laparoscopic partial nephrectomy (LPN) for renal masses in patients.

METHODSFrom January 2010 to December 2012, 39 patients treated with LPN for a single T1 renal tumor were enrolled in this prospective study. There were 24 male and 15 female patients. Their age was (58 ± 10) years old, and their body mass index was (27 ± 3) kg/m(2). The mean operation time was (132 ± 12) minutes, and the mean WIT was (29 ± 8) minutes. Clinical parameters, the single glomerular filtration rates (sGFR) were compared before the operation and after 3 and 12 months in order to observer the effects on renal function and find the factors predicting the renal function impairment.

RESULTSThere were significant differences between 3, 12 months after the operation ((26.8 ± 5.6) ml/min and (28.6 ± 5.6) ml/min, respectively) and preoperation ((31.9 ± 6.3) ml/min) in sGFR (F = 4.882 and 5.511, both P < 0.05). And there were significant negative correlations between the sGFR in 3 and 12 months after the operation and WIT (r = -0.569, P = 0.000 and r = -0.448, P = 0.004) . The preoperative sGFR (β = 0.260, 95%CI:0.089-0.431) and WIT (β = 0.369, 95%CI:0.189-0.555) were independent predictors for function decline of the operated kidney (both P < 0.05). The analysis showed that the effects of WIT within 30 minutes on renal function is relatively small. Longer WIT was associated with lower postoperative sGFR values (F = 22.128 and 20.552, both P = 0.000) .

CONCLUSIONSFor the LPN operation, the longer of the WIT, the more serious of renal function damage. sGFR is an accurate measurement to assess the renal damage. Every effort should be made to minimise WIT during LPN, and the limit of 30 minutes should be not exceeded.

Aged ; Female ; Humans ; Kidney ; physiopathology ; Kidney Function Tests ; Kidney Neoplasms ; surgery ; Laparoscopy ; Male ; Middle Aged ; Nephrectomy ; Operative Time ; Prospective Studies ; Warm Ischemia ; adverse effects

OBJECTIVE: To explore the role of neutrophil-to-lymphocyte ratio（NLR) in patients with newly diagnosed essential thrombocythemia（ET) and its relationship with thrombotic events. METHODS: The clinical and follow-up data of 150 ET patients were retrospectively analyzed. The risk factors of thrombotic events and role of NLR by statistical methods. RESULTS: Age (P<0.01) and JAK2V617F mutation (P<0.01) were the independent risk factors for thrombotic events at diagnosis; WBC count (P<0.05), NLR (P<0.01), age (P<0.05) and thrombosis history at diagnosis (P<0.05) were independent risk factors for future thrombotic events. The ROC curve showed that NLR for prediction of future thrombotic events was suprior to other risk factors. The Kaplan-Meier analysis showed that the progress-free survival time in thrombotic events patients with higher NLR (median survival 22.3 months, 95% CI:17.8-26.8) was significantly shorter than that of patients with lower NLR (median survival 55.5 months, 95% CI:53.4-57.5) . After follow-up 60 months, the thrombosis progress-free survival in lower NLR patients reached 97.4%, while that in the patients with higher NLR was rate 46.7%. CONCLUSION NLR at diagnosis is a better predictive parameter for future thrombotic events than other clinical parameters in ET patients, but without corralations with thrombosis at diagnosis.
Humans ; Lymphocyte Count ; Lymphocytes ; Neutrophils ; Prognosis ; Retrospective Studies ; Thrombocythemia, Essential ; Thrombosis

OBJECTIVETo detect the expression of PTPN11 gene in acute myeloid leukemia (AML) cell line and to explore the effects of PTPN11 over expressing on proliferation and apotosis of AML cell lines.

METHODSThe expression of PTPN11 in AML cell lines（HEL,U937, K562, KG-1, HL -60） was detected by RT-PCR, Q-PCR and Western blot. The PTPN11 gene was amplified by RT-PCR. PTPN11 DNA fragement and the lentiviral vector PCDH-CD513B were digested by BamHI and EcoRI, and then ligated by T4 DNA ligase. Recombinant lentivirus was generated by co-transfection of three-plasmids into 293FT cells using lipofectamine 2000. Then Q-PCR and Western blot were used to detect the expression of PTPN11 in the lentivirus infected HEL and U937 cells. The CCK-8 and Annexin V/7-AAD assays were performed to evaluate effects of PTPN11 on proliferation, apoptosis of HEL and U937 cells.

RESULTSAll 5 AML cell lines expressed the PTPN11 gene, restriction analysis and gene sequencing confirmed that recombinant lentiviral vector was successfully constructed. After transfection of cells with the lentivirus, the recombinant plasmid could stably up-regulate the expression of PTPN11. Analysis of the proliferation and apoptosis of transfected AML cells indicated that as compared with the control group, the OD values of over-expression group were significantly higher and the apoptotic rates were significantly lower (P<0.05).

CONCLUSIONPTPN11 is expressed in all the 5 AML cell lines. The lentiviral expression vector carrying human PTPN11 and the engineered HEL and U937 cell lines stably up-regulating PTPN11 gene expression are successfully obtained. Over-expression of PTPN11 promotes the proliferation of AML cell lines and inhibit then apoptosis.

Apoptosis ; Cell Line, Tumor ; Gene Expression ; Genetic Vectors ; Humans ; Lentivirus ; Leukemia, Myeloid, Acute ; Plasmids ; Protein Tyrosine Phosphatase, Non-Receptor Type 11 ; Transfection