OBJECTIVES: To investigate the protective effect of Yiqi Yangyin Huazhuo Tongluo Formula (YYHT) against high glucose-induced injury in mouse renal podocytes (MPC5 cells) and the possible mechanism. METHODS: Adult Wistar rats were treated with 19, 38, and 76 g/kg YYHT or saline via gavage for 7 days to prepare YYHT-medicated or blank sera for treatment of MPC5 cells cultured in high glucose (30 mmol/L) prior to transfection with a miR-21a-5p inhibitor or a miR-21a-5p mimic. The changes in miR-21a-5p expressions and the mRNA levels of FoxO1, PINK1, and Parkin in the treated cells were detected with qRT-PCR, and the protein levels of nephrin, podocin, FoxO1, PINK1, and Parkin were detected with Western blotting. Autophagic activity in the cells were evaluated with MDC staining. The effect of miR-21a-5p mimic on FoxO1 transcription and the binding of miR-21a-5p to FoxO1 were examined with luciferase reporter gene assay and radioimmunoprecipitation assay. RESULTS: MPC5 cells exposed to high glucose showed significantly increased miR-21a-5p expression, lowered expressions of FoxO1, PINK1, and Parkin1 mRNAs, and reduced levels of FoxO1, PINK1, parkin, nephrin, and podocin proteins and autophagic activity. Treatment of the exposed cells with YYHT-medicated sera and miR-21a-5p inhibitor both significantly enhanced the protein expressions of nephrin and podocin, inhibited the expression of miR-21a-5p, increased the mRNA and protein expressions of FoxO1, PINK1 and Parkin, and upregulated autophagic activity of the cells. Transfection with miR-21a-5p mimic effectively inhibited the transcription of FoxO1 and promoted the binding of miR-21a-5p to FoxO1 in MPC5 cells, and these effects were obviously attenuated by treatment with YYHT-medicated sera. CONCLUSIONS YYHT-medicated sera alleviate high glucose-induced injury in MPC5 cells by regulating miR-21a-5p/FoxO1/PINK1-mediated mitochondrial autophagy.
Animals ; MicroRNAs/genetics* ; Podocytes/pathology* ; Drugs, Chinese Herbal/pharmacology* ; Autophagy/drug effects* ; Rats, Wistar ; Protein Kinases/metabolism* ; Rats ; Forkhead Box Protein O1 ; Mice ; Mitochondria/drug effects* ; Ubiquitin-Protein Ligases/metabolism* ; Glucose ; Diabetic Nephropathies ; Male ; Membrane Proteins/metabolism* ; Intracellular Signaling Peptides and Proteins

Objective:To evaluate the difference in safety and immunogenicity of live rotavirus vaccine (oral) and measles, mumps and rubella (MMR) vaccine immunized alone or in combination.Methods:This study recruited 1 752 children aged 8-9 months who had not been vaccinated with live rotavirus vaccine (oral) or MMR vaccine after birth. The subjects were divided into three groups: study group (652 subjects, immunized with live rotavirus vaccine and MMR vaccine), control group 1 (723 subjects, immunized with live rotavirus vaccine) and control group 2 (377 subjects, immunized with MMR vaccine). Local and systemic adverse reactions within 30 d after vaccination were recorded. Serum samples were collected before and 35-42 d after immunization for analyzing the changes in antibodies.Results:Immunization alone or in combination with live rotavirus vaccine (oral) and MMR vaccine achieved similar results in the positive rates and concentrations of antibodies against rotavirus, measles and rubella viruses ( P>0.05). Moreover, the positive rates and the concentrations of the three antibodies were increased after vaccination. Compared with the control group 2, the concentration of antibody against mumps virus in the study group was increased ( P<0.05), but no significant difference in the positive rate of antibody against mumps virus was found between the two groups ( P>0.05). The positive rate and the concentration of antibody against mumps virus were increased after combined immunization or immunization with MMR vaccine alone. The overall incidence of fever and diarrhea was 1.54% (27/1 752) and 0.63% (11/1 752). No other abnormal reactions, incidental reactions or adverse reactions of any clinical significance were observed. Conclusions:Live rotavirus vaccine (oral) and MMR vaccine immunized alone or in combination showed good immunogenicity and safety.

Objective:To explore the short-term outcomes of reconstruction of tumorous critical bone defects at femoral shaft with a 3D printed ultra-short stem with a porous structure.Methods:From September 2016 to June 2018, 8 patients underwent reconstruction of critical bone defects with a 3D printed ultra-short stem with a porous structure after resection of femoral shaft malignant tumor at Department of Orthopaedics, West China Hospital. There were 4 males and 4 females, with an average age of 36.9 years (from 11 to 61 years). Their preoperative Enneking staging was stage Ⅱb in all. There were 3 osteosarcomas, 2 Ewing sarcomas, 2 chondrosarcomas and one periosteal osteosarcoma. Preoperative CT/MRI image fusion technology was used to define the surgical boundary, design the guide plate and prosthesis, and perform surgical simulation. Tomosynthesis-shimadzu Metal Artefact Reduction technology was used to evaluate osseointegration. Complications and bone oncology prognosis of the patients were documented. The lower limb function of the patients was evaluated using Musculoskeletal Tumor Society (MSTS) 1993 scoring and knee range of motion.Results:The overall follow-up time ranged from 36 to 50 months, averaging 42.8 months. During operation one patient sustained a periprosthesis fracture, the union of which was followed up after wire assisted fixation. There was no local tumor recurrence, lung metastasis or death. The last follow-up revealed good osseointegration and basically isometric lower extremities in all cases. There was no such a complication as aseptic loosening of the prosthesis, deep infection or prosthesis fracture during the follow-up period. At the last follow-up in the 8 patients, the flexion range of the knee joint was 116.2°±9.1°, significantly improved compared with that before operation (98.8°±10.9°), and the MSTS score was (26.2±2.1) points, also significantly improved compared with that before operation [(21.6±1.8) points] ( P<0.05). Conclusions:Reconstruction with a 3D printed ultra-short stem with a porous structure is an accurate operation for femoral shaft tumorous bone defects. With careful preoperative design, intraoperative manipulation and strict postoperative follow-up management, this operation can lead to fine early curative outcomes for long shaft critical bone defects.

Objective:To determine hydrazine quantitatively in workplace air by gas chromatography with large bore capillary column.Methods:In October 2019, hydrazine in the air was adsorbed by acid silica gel tube sampling and desorped using sulfuric acid solution. After derivatization with furfural and extraction, the content of hydrazine was determined by DM-FFAP capillary column gas chromatography with flame ionization detector.Results:The linear regression equation was y=353.8 x+21.2 ( r=0.9998) between 0.1-2.0 μg/ml of target concentration. The detection limit was 0.030 μg/ml. The lower limit of quantification was 0.100 μg/ml. If 15 L air sample was collected, the minimum detection concentration was 0.004 mg/m 3 and the minimum quantitative concentration was 0.013 mg/m 3 respectively. The average desorption efficiency was 86.5%-89.4%. The recovery was 94.4%-97.1%. The relative standard deviation was 1.6%-4.9%. Hydrazine and furfural derivative was 2-furaldehyde hydrazine. Conclusion:The method has symmetrical peak shape of hydrazine derivatives chromatographic peaks, short analysis time, easy operation, and is suitable for the determination of the concentration of hydrazine in the air in the workplace.

Objective:To determine hydrazine quantitatively in workplace air by gas chromatography with large bore capillary column.Methods:In October 2019, hydrazine in the air was adsorbed by acid silica gel tube sampling and desorped using sulfuric acid solution. After derivatization with furfural and extraction, the content of hydrazine was determined by DM-FFAP capillary column gas chromatography with flame ionization detector.Results:The linear regression equation was y=353.8 x+21.2 ( r=0.9998) between 0.1-2.0 μg/ml of target concentration. The detection limit was 0.030 μg/ml. The lower limit of quantification was 0.100 μg/ml. If 15 L air sample was collected, the minimum detection concentration was 0.004 mg/m 3 and the minimum quantitative concentration was 0.013 mg/m 3 respectively. The average desorption efficiency was 86.5%-89.4%. The recovery was 94.4%-97.1%. The relative standard deviation was 1.6%-4.9%. Hydrazine and furfural derivative was 2-furaldehyde hydrazine. Conclusion:The method has symmetrical peak shape of hydrazine derivatives chromatographic peaks, short analysis time, easy operation, and is suitable for the determination of the concentration of hydrazine in the air in the workplace.

Objective:To assess the antibody persistence 3-5 years following vaccination of measles and rubella combined live-attenuated vaccine (MR) at 8 months of age and measles, mumps and rubella combined attenuated live vaccine (MMR) at 18 months of age.Methods:In 2016, 18-month-old children who were vaccinated with one dose of MR vaccine at the age of 8 months were recruited in Hebei Province as group 1; 4-, 5- and 6-year-old children who were vaccinated with one dose of MR vaccine at the age of 8 months and one dose of MMR vaccine at 18 months of age were recruited in Shanxi, Inner Mongolia and Beijing as group 2, group 3 and group 4, respectively. Serum samples were collected to detect IgG antibodies against measles, mumps and rubella by ELISA. Geometric mean concentrations (GMCs) of measles, mumps, and rubella antibodies were compared among groups by analysis of variance or non-parametric test. Seropositive rates were compared among groups by Chi-square test or Fisher′s exact test. Results:A total of 650 children were included in this study. Seropositive rates of measles, mumps and rubella antibodies 30 d after vaccination of 150 18-month-old children with one dose of MMR vaccine were 100%, 91.33% and 100%, respectively, and the GMCs were 1 846.87 mIU/ml, 299.91 IU/ml and 111.33 IU/ml, respectively. Seropositive rates of measles, mumps and rubella antibodies 3-5 years after vaccination one dose of MR vaccine at 8 months of age and one dose of MMR vaccine at 18 months of age were above 94%, 79% and 71%, respectively, and the GMCs were above 830 mIU/ml, 240 IU/ml and 31 IU/ml. No significant difference in the seropositive rates of the three antibodies was observed among groups 2, 3 and 4 ( P>0.05). There was no significant difference in the GMCs of measles or mumps antibodies among the three groups ( P>0.05), but the differences in the GMCs of rubella antibodies were statistically significant ( P=0.034). Conclusions:Measles, mumps and rubella antibodies persisted for 3-5 years without significant decrease after vaccination one dose of MR vaccine at 8 months of age and one dose of MMR vaccine at 18 months of age.

Objective: To evaluate the immunogenicity and safety of concomitant administration of 23-valent pneumococcal polysaccharide vaccine (PPV23) and trivalent influenza vaccine (TIV) in preschoolers. Methods: A total of 1 035 children aged 3-7 years were enrolled in this study and randomly divided into three groups, inoculated PPV23, TIV and both, respectively. A one-year follow-up study was conducted for immunogenicity and safety analysis. Results: A total of 1 035 serological specimens were collected, including 327 in PPV23 group, 348 in TIV group and 360 in concomitant vaccination group. No significant differences in geometric mean concentrations (GMC) of seven pneumococcal serotypes were observed between the PPV23 group and the concomitant vaccination group. Compared with the TIV group, the concomitant vaccination group showed higher serological conversion rate of H3 type (88.75% vs 84.20%, P=0.01), but lower serological conversion rate of B type (92.84% vs 98.56%, P<0.001). There was no significant difference in the primary adverse reactions between the three groups (P=0.197). The rate of secondary adverse reactions occurred in the concomitant vaccination group was 3.61%, which was higher than that of the other two groups (both P<0.001). All adverse reactions were mild or moderate, and cured after treatment. Conclusions Concomitant immunization with PPV23 and TIV is safe and have good immunogenicity, thus a viable immune strategy for susceptible children.

Objective To investigate the effect of low salt diet on vascular remodeling of rat induced by high fructose(HF).Methods Wistar male rats weighed 180-200 g were fed for 8 weeks and randomly divided into 6 groups:(1) control group was given the normal fodder and distilled water;(2) high fructose group(HF) was given normal fodder (0.5 ％ NaCl,w/w) and fructose water(10 ％,w/v);(3) high-salt group (HNa) was given high salt fodder (7 ％ NaCl,w/w) and distilled water;(4) high fructose combined with high salt diet group(HFNa) was simultaneously given high salt fodder and 10 ％ fructose water;(5)high fructose combined low salt group(HFLNa) was simultaneously given low salt fodder and 10％ fructose water;(6) high fructose combined with spirotaclone group(HFE) was given 10％ fructose water for 4 weeks and then added with spirotaelone(50 mg · kg-1 · d-1 by tube feeding) for continuous 4 weeks.The changes of arterial blood pressure,vascular wall histological evaluation and expression of α-SMA and fibronectin in vascular wall were detected in each group.Results (1) Compared with the blood pressure[(111.03 ±9.17) mm Hg] in the control group,the blood pressure in the HF and HNa groups were (133.94± 5.86) mm Hg and (128.09±7.56) mm Hg respectively,which were significantly increased(P＜0.05);(2) HF mainly caused the hyperplasia of vascular wall middle layer smooth muscle.The a-SMA expression results in the HF group was (0.006 3 ±0.000 21),which in the control group was (0.004 6 ± 0.000 31),the difference was statistically significant(P＜0.05),moreover which promoted the elastic fibers increase;while HNa mainly stimulated the elastic fibers to thicken and extracellular matrix deposition,the fibronectin expression was 0.002 6 ± 0.000 2 in the HNa group and (0.004 7±0.000 2)in the HF group,compared with(0.001 3±0.000 1)in the normal group,which were significantly increased(P＜0.001);(3) the blood pressure was (106.04±9.59) mm Hg in the HFLNa group,(103.99±7.12) mm Hg in the HFE group,compared with(133.94±5.86) mm Hg in the HF group,showing that the blood pressure in the HFLNa group and HFE group was significantly decreased compared with the HF group (P＜0.05);moreover the vascular remodeling in the HFLNa group(0.006 8±0.000 2) and HFE group (0.004 2±0.000 4) was improved,and compared with the HF group(0.006 3±0.000 2),α-SMA expression was significantly decreased (P＜0.05).Conclusion Low salt diet can effectively improve vascular remodeling induced by HEF.

Objective: To establish a method to detect the concentrations of ethylenediamine in the air of workplace by Ion chromatography. Methods: Ethylenediamine in the air of workplace adsorpted by Silicone tube, then ultrasonic desorped by Sulfuric acid solution (0.1 mol/L) , the desorption solution qualitative and quantitative determination by Ion chromatography. Results: For ethylenediamine, the calibration curves were liner in the range of 0.1-100.0 μg/ml, the method detection limit is 0.1 μg/ml, the minimum detectable concentration is 0.13 mg/m³ (in terms of sampling 7.5 L) . The desorption efficiency is more than 90% on spiked levels of 2.0, 4.0 and 8.0 μg/ml, the average desorption efficiency of the method was 96.1%, the precision was 1.8%-2.1%. The recovery rate of the added standard is 98.5%-104.2%. Conclusion The method simplifies the preprocessing steps and improves the sensitivity. All the indexes are in conformity with the requirements of the method formulation guide. It is suitable for the determination of ethylenediamine in the air of the workplace.

OBJECTIVETo observe the effects of conventional western medication and joss stick moxibustion combined with pricking and cupping for herpes zoster in acute stage, and to explore its analgesic mechanism.

METHODSSeventy patients with acute herpes zoster were randomized into an observation group (33 cases after 2 dropping) and a control group (34 cases after 1 dropping). Patients in the observation group were treated with joss stick moxibustion combined with pricking and cupping at localpoints for 7 times, once every other day. Oral acyclovir, vitamin Band mecobalamin tablets were applied in the control group for continuous 14 days, and interferon injection was used for continuous 6 days, etc. The herpes evaluation indexes of blister stopping time, scab time and decrustation time as well as pain intensity were observed before and after treatment. Peripheral serum substance P (SP) content of herpes local situation was detected. The comprehensive effects were evaluated.

RESULTSThe blister stopping time, scab time and decrustation time in the observation group were shorter than those in the control group (all<0.05). There was no statistical significance for pain relief degree between the two groups (>0.05). The pain beginning to ease time and duration time in the observation group were better than those in the control group (both<0.05). The contents of SP in the two groups decreased after treatment (both<0.01), and it was better in the observation group (<0.05). The total effective rate of the observation group after treatment was 87.9% (29/33), and that of the control group was 85.3% (29/34), which were not statistically significant (>0.05). The cured rate of the observation group was better than that of the control group [66.7% (22/33) vs 58.8% (20/34),<0.05].

CONCLUSIONJoss stick moxibustion combined with pricking and cupping are effective for herpes zoster, which have quicker and good analgesic effects than conventional western medication. Its mechanism may be related to reducing the content of SP more fast and to a larger degree.