Objective:To evaluate endoscopic retrograde appendicitis therapy in treatment of children with acute uncomplicated appendicitis.Methods:Sixty children patients were admitted at the Affiliated Hospital of Zhengzhou University from Oct 2019 to Jun 2021 and were divided into ERAT group ( n=30) and LA group ( n=30). Results:All operations were successfully performed . ERAT children started oral feeding earlier [(6.8±2.0) h vs. (12.3±2.0) h, t=-10.636, P<0.001], postoperative hospital stay was shorter [(3.2±1.3) d vs. (5.0±1.3) d, t=-5.360, P<0.001]. After 14 months follow up, the recurrence rate in the ERAT group was 6%. The complication rate of LA was 10%. Conclusion:ERAT is a safe and effective therapy in treating children with acute uncomplicated appendicitis with low,acceptable recurrence rate.

Two male patients(patient 1,64 years old;patient 2,63 years old)received parenteral nutrition in fasting state due to the future surgical operation for gastric space-occupying lesions and gastrointestinal bleeding,respectively.Both of them developed fatigue and cold sweat near the end of the first Ⅳ infusion of the lipid emulsion (10％)/amino acids (15) and glucose (20％) injection.Their instant blood glucose levels were 2.2 mmol/L and 2.1 mmol/L,respectively.The symptoms disappeared about 10 minutes after the immediate discontinuation of the injection and an intravenous injection of 50％ glucose injection 40 ml.And their instant blood glucose levels were 5.6 mmol/L and 5.4 mmol/L,respectively.Patient 2 developed the above symptoms again near the end of the second infusion of the drug next day and his instant blood glucose level was 2.6 mmol/L.His symptoms disappeared again after the drug withdrawal and treatment of an intravenous injection of 50％ glucose 40 ml and an Ⅳ infusion of 10％ glucose 250 ml for about 15 minutes.Then his instant blood glucose level was 5.9 mmol/L.

Two male patients(patient 1,64 years old;patient 2,63 years old)received parenteral nutrition in fasting state due to the future surgical operation for gastric space-occupying lesions and gastrointestinal bleeding,respectively.Both of them developed fatigue and cold sweat near the end of the first Ⅳ infusion of the lipid emulsion (10％)/amino acids (15) and glucose (20％) injection.Their instant blood glucose levels were 2.2 mmol/L and 2.1 mmol/L,respectively.The symptoms disappeared about 10 minutes after the immediate discontinuation of the injection and an intravenous injection of 50％ glucose injection 40 ml.And their instant blood glucose levels were 5.6 mmol/L and 5.4 mmol/L,respectively.Patient 2 developed the above symptoms again near the end of the second infusion of the drug next day and his instant blood glucose level was 2.6 mmol/L.His symptoms disappeared again after the drug withdrawal and treatment of an intravenous injection of 50％ glucose 40 ml and an Ⅳ infusion of 10％ glucose 250 ml for about 15 minutes.Then his instant blood glucose level was 5.9 mmol/L.

Objective To estimate the influence of palmitic acid (PA) on the proliferation of and production of inflammatory mediators by a human keratinocyte line HaCaT.Methods Cultured HaCaT cells were treated with PA of eight concentrations (0-200 μmol/L) for 3-24 hours followed by the evaluation of cell proliferation by using the cell counting kit-8.According to the proliferation assay,four concentrations (75,100,125,150 μmol/L) of PA were selected and used to treat HaCaT cells for 24 hours,then,fluorescence-based immunohistochemical staining was performed to observe the nuclear translocation of nuclear factor (NF)-κB p65,enzyme linked immunosorbent assay (ELISA) to determine the level of interleukin (IL)-6 in the supernatant of culture medium,real-time PCR to detect the mRNA expressions of peroxisome proliferator-activated receptor oα (PPARα) and IL-6,and Western blot to quantify the protein expressions of PPARα as well as total and nuclear NF-κB p65.Those HaCaT cells receiving no treatment served as the control group.Statistical analysis was carried out by one-factor analysis of variance using the GraphPad Prism 5.0 software.Results The HaCaT cells treated with PA of 50-175 μ mol/L showed accelerated proliferation compared with the control HaCaT cells (all P ＜ 0.05).PA from 75 to 150 μmol/L enhanced the nuclear translocation of NF-κB p65,mRNA and protein expressions of PPARα,as well as the mRNA expression and supernatant level of IL-6 in a dose-dependent manner.The relative expression level of nuclear NF-κB p65 protein was 0.4536 ± 0.0173,0.5184 ± 0.0206,0.5333 ± 0.0231,0.6160 ± 0.0297,and the supernatant level of IL-6 was (31.5677 ± 0.2268),(32.3773 ± 0.4156),(32.9837 ± 0.0029) and (33.6890 ± 0.0936) ng/L,in HaCaT cells treated with PA of 75,100,125 and 150 μmol/L,respectively,compared to 0.3237 ± 0.0114 (all P ＜ 0.01) and (30.4577 ± 0.5131) ng/L (all P ＜ 0.01) in the control HaCaT cells,respectively.Conclusions PA can accelerate the proliferation of HaCaT cells,enhance NF-κB nuclear transfer,PPARα expression and IL-6 secretion in a dose-dependent manner within a certain concentration range,and may exert a promoting role in the activation and expression of some inflammatory factors.

Objective To observe the effect of sirolimus,an autophagy enhancer,on premature senescence in fibroblasts induced by repeated exposure to a subtoxic dose of ultraviolet B (UVB).Methods Skin fibroblasts from foreskin tissue of healthy adolescents were classified into six groups:control group cultured in Dulbecco's modified Eagles' medium (DMEM) containing 1％ calf serum,UVB group receiving UVB irradiation only,sirolimus group treated with sirolimus of 10 mg/L (added after daily exchange of culture medium),and three combined groups receiving UVB irradiation immediately followed by overnight treatment with sirolimus of 0.1,1.0 and 10.0 mg/L respectively.UVB irradiation was given at a dose of 10 mJ/cm2 once a day for five successive days.After five days of treatment,cell counting kit-8 (CCK-8) was used to evaluate cell viability,β-galactosidase staining to detect senescent ceils,Western blot to quantify the expressions of p53,LC3-B and beclin 1 in these fibroblasts.Autophagy level was determined by acridine orange staining followed by fluorescence microscopy and transmission electron microscopy.Data were processed by the SPSS 16.0 software,and statistical analysis was done by one-way analysis of variance,t test and least significance difference.Results Sirolimus significantly increased the proliferative activity of fibroblasts in a dose-dependent manner,with the absorbance value at 450 nm being 0.27 ± 0.02,0.36 ± 0.04 and 0.39 ± 0.04 for fibroblasts irradiated with UVB followed by treatment with sirolimus of 0.1,1.0 and 10 mg/L respectively,compared to 0.26 + 0.01 for fibroblasts irradiated with UVB only (all P ＜ 0.05).Significant differences were also observed between the fibroblasts irradiated with UVB followed by treatment with sirolimus of 0.1,1.0 and 10 mg/L and those irradiated with UVB only in the percentage of β-galactosidasepositive fibroblasts (92.50％ ± 0.34％,42.40％ ± 0.53％ and 6.20％ ± 0.39％ vs.95.10％ ± 0.32％,all P ＜ 0.05)and intracellular intensity of acridine orange-induced fluorescence (36.43 ± 0.24,45.25 ± 0.33 and 48.69 ± 0.37 vs.33.99 ± 0.32,all P ＜ 0.05).Moreover,the expressions of p53,LC3-B and beclin 1 in the three combined groups differed significantly from those in the UVB group (all P ＜ 0.05).Conclusion Sirolimus can inhibit UVBinduced premature senescence likely via upregulation of autophagy in fibroblasts.

Obective To evaluate the effects of dihydrotestosterone (DHT) on the expression of sterol regulatory element-binding protein-1c (SREBP-1c) in human HaCaT keratinocytes.Methods HaCaT cells were cultured in vitro and classified into 4 groups,i.e.,control group receiving no treatment,DIIT group treated with 3 different concentrations (10,100,1000 nmol/L) of DHT,LY294002 plus DHT group treated with DHT of 100 nmol/L after 40-minute pretreatment with the PI3K inhibitor LY294002 of 50 μmol/L,PD98059 plus DHT group treated with DHT of 100 nmol/L after 40-minute pretreatment with the MEK inhibitor PD98059 of 50 μmol/L.After another 24-hour culture,real time PCR and Western blot were carried out to detect the expression of SREBP-1c mRNA and protein in HaCaT cells,respectively.Western blot was also performed to determine the phosphorylation levels of protein kinase B (AKT),extracellular signal-regulated kinase (ERK),p38 mitogen activated protein kinase and c-Jun N-terminal kinase (JNK) in the HaCaT cells.Results DHT could enhance the expression of SREBP-1c mRNA and protein in HaCaT cells in a concentration-dependent manner,and induce the phosphorylation of AKT and ERK,but not that of P38 or JNK.The expressions of SREBP-1c mRNA and protein were significantly decreased in HaCaT cells treated with LY294002 plus DHT (7.4780 ± 1.2638 vs.21.6170 ± 2.2759,t =9.406,P ＜ 0.05; 0.7113 + 0.0313 vs.2.2577 + 0.0601,t =39.498,P ＜ 0.05),but experienced no statistical changes in those treated with PD98059 and DHT(both P ＞ 0.05),compared with those treated with DHT only.Conclusion DHT can induce the expression of SREBP-1c mRNA and protein in HaCaT cells,likely via the PI3K/AKT signaling pathway.

Objective To detect the expression of translationally controlled tumor protein (TCTP) in squamous cell carcinoma (SCC) tissue and cell lines A431 and SCL-1,and to evaluate the effect of TCTP on apoptosis in and proliferation of SCC cells.Methods An immunohistochemical method was used to measure the expression of TCTP in tissue specimens from 65 patients with SCC.Western blot was performed to detect the expression of TCTP in A431 and SCL-1 cells.Three small interference RNAs (siRNAs) targeting the TPT1 gene were designed,synthesized,and transfected into A431 cells.Then,reverse transcription (RT)-PCR and Western blot were conducted to measure the expression of TPT1 mRNA and TCTP,respectively,methyl thiazolyl tetrazolium (MTT) assay and flow cytometry were carried out to detect cell proliferation and apoptosis,respectively.Results TCTP was overexpressed in SCC tissue specimens,and the expression level was positively correlated with the histologic grading of SCC (P ＜ 0.05).Western blot showed that TCTP was expressed in both A431 and SCL-1 cells,and the expression was relatively high in A431 cells.The transfection efficiency of siRNAs varied from 90％ to 95％.A decrease in the expression of TPT1 mRNA and TCTP was induced by the siRNAs in A375 cells (all P ＜ 0.05).Conclusion The downregulation of TCTP expression may increase the apoptosis in and suppress the proliferation of A431 cells.

ObjectiveTo observe the effect of topical sulfotanshinone sodium(STS) on sebaceous hyperplasia in animal models. MethodsThe sebaceous gland spots of adult male Syrian hamster flank organ served as the animal model. Sulfotanshinone sodium(0.5％) was applied to sebaceous gland spots in the right flank organ thrice daily, while those in the left were treated with normal saline as control. Parameters were examinedbefore, 10 days, 20 days and 30 days after the beginning of the topical treatment. A vernier caliper was utilized to measure the size of sebaceous gland spots, hematoxylin and eosin(HE) staining to observe the structure of sebaceous glands, immunohistochemistry to determine the expression of proliferating cell nuclear antigen (PCNA) in sebaceous gland cells, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay to assess the apoptosis of sebaceous gland cells. ResultsAt the baseline, no significant difference was observed in the size of sebaceous gland spots or in the proliferation and apoptosis of sebaceous gland cells between the two sides of flank organ(all P ＞ 0.05), with tightly arranged intact sebaceous glands. Compared with normal saline, sulfotanshinone sodium significantly reduced the size of sebaceous gland spots(P ＜ 0.05). Sebaceous glands were loosely arranged with decreased quantity and volume and obviously atrophic on day 30 in the right flank organ of hamsters. A decrease was observed in the expression of PCNA in sulfotanshinone sodium treated sebaceous gland cells compared with those treated with normal saline(P ＜ 0.01 ), which was more striking on day 10 and 20(both P ＜ 0.005). Sulfotanshinone sodium also induced an enhancement of apoptosis in sebaceous gland cells (P ＜ 0.01 ), which was more apparent on day 20 (P ＜ 0.005 ), and the degree of apoptosis was higher in the central area than in the peripheral area of sebaceous glands. ConclusionSulfotanshinone sodium can reduce the size and alter the microstructure of sebaceous gland spots, and inhibit the hyperplasia of sebaceous glands.