Overexpressed SIRT6 ameliorates doxorubicin-induced cardiotoxicity and potentiates the therapeutic efficacy through metabolic remodeling

Since the utilization of anthracyclines in cancer therapy, severe cardiotoxicity has become a major obstacle. The major challenge in treating cancer patients with anthracyclines is minimizing cardiotoxicity without compromising antitumor efficacy. Herein, histone deacetylase SIRT6 expression was reduced in plasma of patients treated with anthracyclines-based chemotherapy regimens. Furthermore, overexpression of SIRT6 alleviated doxorubicin-induced cytotoxicity in cardiomyocytes, and potentiated cytotoxicity of doxorubicin in multiple cancer cell lines. Moreover, SIRT6 overexpression ameliorated doxorubicin-induced cardiotoxicity and potentiated antitumor efficacy of doxorubicin in mice, suggesting that SIRT6 overexpression could be an adjunctive therapeutic strategy during doxorubicin treatment. Mechanistically, doxorubicin-impaired mitochondria led to decreased mitochondrial respiration and ATP production. And SIRT6 enhanced mitochondrial biogenesis and mitophagy by deacetylating and inhibiting Sgk1. Thus, SIRT6 overexpression coordinated metabolic remodeling from glycolysis to mitochondrial respiration during doxorubicin treatment, which was more conducive to cardiomyocyte metabolism, thus protecting cardiomyocytes but not cancer cells against doxorubicin-induced energy deficiency. In addition, ellagic acid, a natural compound that activates SIRT6, alleviated doxorubicin-induced cardiotoxicity and enhanced doxorubicin-mediated tumor regression in tumor-bearing mice. These findings provide a preclinical rationale for preventing cardiotoxicity by activating SIRT6 in cancer patients undergoing chemotherapy, but also advancing the understanding of the crucial role of SIRT6 in mitochondrial homeostasis.

Ample dietary fat reduced the risk of primary vesical calculi by inducing macrophages to engulf budding crystals in mice

Although primary vesical calculi is an ancient disease, the mechanism of calculi formation remains unclear. In this study, we established a novel primary vesical calculi model with d,l-choline tartrate in mice. Compared with commonly used melamine and ethylene glycol models, our model was the only approach that induced vesical calculi without causing kidney injury. Previous studies suggest that proteins in the daily diet are the main contributors to the prevention of vesical calculi, yet the effect of fat is overlooked. To assay the relationship of dietary fat with the formation of primary vesical calculi, d,l-choline tartrate-treated mice were fed a high-fat, low-fat, or normal-fat diet. Genetic changes in the mouse bladder were detected with transcriptome analysis. A high-fat diet remarkably reduced the morbidity of primary vesical calculi. Higher fatty acid levels in serum and urine were observed in the high-fat diet group, and more intact epithelia in bladder were observed in the same group compared with the normal- and low-fat diet groups, suggesting the protective effect of fatty acids on bladder epithelia to maintain its normal histological structure. Transcriptome analysis revealed that the macrophage differentiation-related gene C-X-C motif chemokine ligand 14 (Cxcl14) was upregulated in the bladders of high-fat diet-fed mice compared with those of normal- or low-fat diet-fed mice, which was consistent with histological observations. The expression of CXCL14 significantly increased in the bladder in the high-fat diet group. CXCL14 enhanced the recruitment of macrophages to the crystal nucleus and induced the transformation of M2 macrophages, which led to phagocytosis of budding crystals and prevented accumulation of calculi. In human bladder epithelia (HCV-29) cells, high fatty acid supplementation significantly increased the expression of CXCL14. Dietary fat is essential for the maintenance of physiological functions of the bladder and for the prevention of primary vesical calculi, which provides new ideas for the reduction of morbidity of primary vesical calculi.

Discovery of highly selective and orally available benzimidazole-based phosphodiesterase 10 inhibitors with improved solubility and pharmacokinetic properties for treatment of pulmonary arterial hypertension

Optimization efforts were devoted to discover novel PDE10A inhibitors in order to improve solubility and pharmacokinetics properties for a long-term therapy against pulmonary arterial hypertension (PAH) starting from the previously synthesized inhibitor

Clinical pathologic analysis on 3 724 cases of salivary gland tumors / 中华口腔医学杂志

Objective@#To investigate the incidence and proportion of salivary gland tumors in order to provide new thinking for clinical diagnosis and treatment.@*Methods@#Collected 3 724 cases salivary gland tumors diagnosed by Pathology Department of Hospital of Stomatology, Jilin University from January 1961 to December 2016. The pathological diagnosis referred to the fourth edition of head and neck-salivary gland tumor histopathological classification standard of WHO. The database was established with Microsoft Excel and analyzed with SPSS 18.0. Made a retrospective analysis and comparison on the numbers of all cases in terms of types, site, gender and age and estimate the trend with the time interval of 8 years, and then make a judgement of the trend of salivary tumors.@*Results@#The benign tumors were more common than the malignant among all periods, the proportion of all tumors was about 2.92∶1; The top three benign tumors were polymorphous adenoma [73.78% (2 046/2 773)], Warthin tumor [15.80% (438/2 773)] and base cell adenoma [8.37% (232/2 773)]. Polymorphous adenoma took up 54.94% (2 046/3 724) of all tumors. The top three malignant tumors were mucous epidermoid carcinoma [31.44% (299/951)], adenoid cystic carcinoma [26.92% (256/951)] and adenocarcinoma [11.88% (113/951)]. As for sex, male female ratio was 0.83∶1. As for site, the pathogenic site of tumors was mainly in parotid gland [63.75% (2 374/3 724)], followed by palatal gland [16.50% (615/3 724)], then submandibular gland [12.67% (472/3 724)]; As for age, the common age was between 51 and 60 years old [23.74% (884/3 724)], followed by 41 to 50 years old [21.56%(803/3 724)].@*Conclusions@#The incidence of benign and malignant salivary gland tumor increased in the 56 years. Females showed a higher incidence. The majority tumors occurred in parotid gland. The most common salivary gland tumor was pleomorphic adenoma and the most common malignant tumor was mucous epidermoid carcinoma. The most common age was in 51-60 years old period.

Maxillary first molar with a single buccal root / 实用口腔医学杂志

Maxillary first molar demonstrates considerable anatomic complexities and abnormalities with respect to the number of roots and root canals. The occurrence of maxillary first molar with a single buccal root is rarely reported in literature. This is a case report of maxillary first molar with a single buccal root and a palatal root,each of which has one canal.

Investigation into the nutritional knowledge, attitude and practices among officers and soldiers living at high altitude and its influential factors / 解放军医学杂志

Objective To investigate the levels of nutritional knowledge, attitude and dietary habits and its influential factors of the officers and soldiers in frontier forces to provide the theory basis for nutrition education.Methods Using self-designed nutritional knowledge-altitude-practice (KAP) questionnaires to survey 90 officers and soldiers who lived in plateau for at least 6 months. Questionnaires were handed out and handed over on the spot. According toMilitary Nutrition Survey and Evaluation Methods (GJB 1636A-2016), we measured the physical index of officers and soldiers, including height, weight and skin thickness of triceps. And we also evaluated body mass index (BMI) and upper arm muscle circumference.Results Nutrition knowledge, attitude and practice conditions of plateau forces were at the average, better and average levels respectively. Nutritional knowledge was significantly positively related to the practices (r=0.283,P=0.003), and was also significantly positively related to the age of the soldiers (r=0.228,P=0.04). Nutritional attitude was positively related to source of officers and soldiers (r=0.339,P=0.035), and nutritional practices were positively related to the upper arm muscle circumference (r=0.222,P=0.030). 65.6% of the officers and soldiers would like to know the knowledge about relationship between diet and disease. 25.6% of the officers and soldiers in the plateau were alcohol users.Conclusions Although the nutrition knowledge of plateau forces were poor, and the nutritional practices were influenced by nutritional knowledge, but the soldiers' attitude of changing unhealthy dietary habits was very positive. It is necessary to undertake the dietary nutrition education related to the high altitude nutrition among plateau forces to guild their nutritional practice scientifically.

An in situ gel system for nasal delivery of menthol: preparation and safety evaluation / 药学实践杂志

Objective To prepare an in situ gel system for nasal delivery of menthol and to evaluate the safety of this formulation.Methods Menthol in situ gel was prepared with deacetylatedgellan gum.The nasal mucocilia toxicities of this formulation was evaluated using in situ toad palate model.Guinea pig skin sensitization test and the rabbit skin irritation test were conducted.Skin allergy and irritation reaction were monitored and scored.Results No significant effect on nasal mucosa ciliary movement and the morphology of rat nasal mucosa were observed.The formulation did not induce any dermal irritation in rabbits.Skin allergic reaction was not found in guinea pigs.Conclusion The preparation of menthol in situ nasal gel with low ciliary toxicity was easily achieved.This gel has good physiological flexibility.The further investigation was warranted for this formulation as an intranasal drug delivery system.

Over-expression and suppression of HMGBI affect proliferation and invasion of endometrial carcinoma and underlying mechanisms / 中南大学学报(医学版)

Objective:To investigate effects of over-expression and suppression of HMGB1 on proliferation and invasion of endometrial carcinoma HEC-1A cell and underlying mechanisms.Methods:Over-expression or silence of HMGB 1 in HEC-1A cell lines were established by lentiviral vector containing HMGB 1 recombinant plasmid or by HMGB 1 shRNA,respectively.Cell counting kit-8,Transwell,and wound healing assay were used to analyze proliferation,invasion,and migration of HEC-1A cells,respectively.Western blot and reverse transcription-PCR (RT-PCR) were used to detect the expression of NF-κB,VEGF,and matrix metalloproteinase 2 (MMP2) in the cells.Results:Over-expression of HMGB1 promoted the proliferation,invasion,and migration of HEC1A cell,and up-regulated NF-κB,VEGF,and MMP2 expressions,while suppression of HMGB1 inhibited the proliferation,invasion,and migration of HEC-1A cells and down-regulated NF-κB,VEGF,and MMP2 expressions.Conclusion:HMGB1 plays an important role in proliferation,invasion,and migration of HEC1A cell via NF-κB/VEGF/MMP2 pathway.HMGB 1 might be a potential target for endometrial carcinoma therapy.

Dietary nutrition status of soldiers living at high altitude / 军事医学

Objective To investigate the dietary nutrition status of plateau frontier forces and to make recommendations on food and nutrients intake for plateau forces.Methods The dietary status of the plateau forces located at an altitude of 4600 m and 4030 m was investigated by weighing method.The dietary ration for soldiers, their daily dietary allowance and dietary balance index were used to evaluate their dietary patterns and nutrient intake.The concentrations of serum vitamin A and 25-OH vitamin D(25-OH VD) were detected by high performance liquid chromatography (HPLC).The arm muscle circumference and body fat rate of soldiers were evaluated by GJB 1636A-2016 and body composition related standard respectively.Results The intake of eggs, milk and plant oil of Unit A was sufficient, while the rest of the food intake did not reach the military standard;The intake of grain and plant oil of Unit B was sufficient,but the rest of the food intake was insufficient.The intake of protein,calcium,phosphonium,sodium,iron,selenium,iodine,copper,manganese,vitamin E and vitamin B3 of Unit A were adequate, but that of energy,potassium,zinc,magnesium,vitamin D,vitamin C,vitamin B1 and vitamin B2 was insufficient, and the intake of vitamin A,vitamin B6,vitamin B9 and vitamin B12 of Unit A was deficient.The intake of energy, protein,vitamin E,vitamin C,vitamin B1 and vitamin B3,and most of the minerals was adequate in Unit B, but vitamin B2 was insufficient.Furthermore, the intake of calcium,iodine,vitamin A,vitamin D and vitamin B was deficient.The proportion of nutrients which supply energy and the energy distribution of three meals in the two units were imperfect.The concentration of plasma vitamin A in both units was sufficient,but the concentration of plasma 25-OH VD was deficient.93.5% soldiers of unit A and 97.7% soldiers of unit B reached the standard of proper muscle circumference,80.6% soldiers of unit A and 70.5% soldiers of unit B had a low body fat rate.Conclusion The dietary structure of plateau border forces is not balanced.The intake of some foods and nutrients is insufficient so that nutritional education is badly needed to improve the dietary status of plateau forces.

Expression of D J-1 protein in endometrial carcinoma tissue and its correlation with prognosis / 中国医师杂志

Objective To investigate the expression and clinical significance of DJ-1 protein in the endometrial carcinoma and delineate its functional roles in endometrial carcinoma progression.Methods Expression of DJ-1 was examined with immunohistochemical staining of 69 paraffin-embedded endometrial carcinoma samples who underwent surgery at Hunan province Tumor Hospital.Kaplan-Meier analysis was performed to describe the survival curve.Univariate and multivariate analyses were performed with COX proportional hazard model.Results (1) The expression of DJ-1 protein in endometrial carcinoma was significantly higher than that in normal endometrial tissues (P ＜ 0.05).(2) There was positive correlation between the high expression of DJ-1 protein in endometrial carcinoma samples and the pathological grade,the International Federation of Gynecology and Obstetrics (FIGO) stage,depth of myometrial invasion,or lymph node metastasis (P ＜ 0.05).(3) Kaplan-Meier analysis indicated that the cumulative 5-year survival rate in the low-DJ-1 expression group was 80.8％,whereas it was only 51.2％ in the high-DJ-1 expression group (P ＜ O.05).COX analysis indicated that the expression of DJ-1 was not the endometrial carcinoma independent prognostic factors for overall survival time (P ＞ 0.05).Conclusions The high expression of DJ-1 protein in endometrial carcinoma samples demonstrated a significantly positive correlation with the pathological grade,FIGO stage,depth of myometrial invasion,and lymph node metastasis.However,DJ-1 is not enough to be independent prognostic factor for overall survival time.

Modeling and analysis of triage in disaster rescue action executed by mobile medical unit using stochastic Petri net / 军事医学

Objective To analyze the process of triage in disaster rescue action performed by a mobile medical unit so that the rescue process can be improved , the efficiency of rescue enhanced , and the decision on health service in rescue action is supported.Methods The process of triage in disaster rescue action was modeled based on stochastic Petri net while the performance of the model was analyzed quantitatively .Results and Conclusion The critical factor which affects the efficiency of rescue work is obtained by analyzing the performance of the model .

Structural modeling and dynamic evolution analysis of hypoxia response networks / 军事医学

Objective To construct an executable model of a hypoxia response network (HRN) and to analyze the dynamic evolution mechanism of an HRN including randomness as well as concurrency based on computer simulation. Methods Specific evolution rules and Gillespie algorithm were adoped to study the dynamic evolution of the structural model based on the construction of a structural model of an HRN using stochastic Petri net ( SPN ) .Results Dynamic evolution laws of an HRN were obtained and the simulation results were consistent with laboratory results in response to descript switch-like behavior of an HRN .Conclusion A visualization model of the HRN can be achieved using SPN method.Simulation results achieved by executing the model based on stochastic simulation using specific kinetic parameters can serve as a nice complement to traditional laboratory results , which can help shed light on the structure and function characteristics of an HRN.

The clinical significance of estradiol gel applying to the therapy of operation combined with GnRHa in endometriosis / 中国医师杂志

Objective To investigate the efficacy and tolerability of estradiol (E2) gel with postoperative patients of moderately severe ovarian endometriosis cyst,who suffer from the side effect of GnRH agonist.Methods Sixty samples were selected with stage Ⅲ ～ Ⅳ ovarian endometriosis after conservative operation by the same operator at the Department of Gynecology,Xiangya Hospital,Central South University,between February 2013 and August 2014.All of the patients after surgeries were randomly divided into three groups:control group (n =20) with only goserelin injection after conservative surgery,treatment group A (n =20) who received estradiol gel,and treatment group B (n =20) who received tibolone as an add-back therapy.The serum E2,follicle stimulating hormone (FSH),luteinizing hormone (LH),hypoestrogenic symptoms,the incidence of uterine bleeding,and the recurrence after conservative operation were compared among three groups.Results Three months later aster treatment with goserelin,the serum level of E2 of three groups showed statistically significant difference (P ＜ 0.05).Serum E2 level of treatment group A was not significantly higher than treatment group B (P ＞ 0.05).The kupperman sore,the incidence of hot flashes sweating,tiring,and agrypnia of treatment groups A and B were significantly lower than the control group (P ＜ 0.05).The incidence of uterine bleeding and recurrence dont differ sharply among three groups (P ＞ 0.05).Conclusions GnRHa combined with estradiol gel can improve symptoms of postoperative patients,reduce side effects effectively,and avoid increase of the risk of vagina bleeding and relapse.

A cost-effectiveness analysis on universal infant rotavirus vaccination strategy in China / 中华流行病学杂志

Objective To evaluate the cost-effectiveness of current universal infant rotavirus vaccination strategy,in China.Methods Through constructing decision tree-Markov model,we simulated rotavirus diarrhea associated cost and health outcome on those newborns in 2012 regarding different vaccination programs as:group with no vaccination,Rotavirus vaccination group and Rotateq vaccination group,respectively.We determined the optimal program,based on the comparison between incremental cost-effectiveness ratio (ICER) and China' s 2012 per capital gross domestic product (GDP).Results Compared with non-vaccination group,the Rotavirus vaccination and Rotateq vaccination groups had to pay 3 760 Yuan and 7 578 Yuan (both less than 2012 GDP per capital) to avert one disability adjusted life years (DALY) loss,respectively.Results from sensitivity analysis indicated that both results were robust.Compared with Rotavirus vaccination program,the Rotateq vaccination program had to pay extra 81 068 Yuan (between 1 and 3 times GDP per capital) to avert one DALY loss.Data from the sensitivity analysis indicated that the result was not robust.Conclusion From the perspective of health economics,both two-dose Rotarix vaccine and three-dose's Rotateq vaccine programs were highly cost-effective,when compared to the non-vaccination program.It was appropriate to integrate rotavirus vaccine into the routine immunization program.Considering the large amount of extra cost that had to spend on Rotateq vaccination program,results from the sensitivity analysis showed that it was not robust.Rotateq vaccine required one more dose than the Rotarix vaccine,to be effective.However,it appeared more difficult to practice,suggesting that it was better to choose the Rotarix vaccine,at current stage.

A cost-effectiveness analysis on universal infant rotavirus vaccination strategy in China / 中华流行病学杂志

Objective To evaluate the cost-effectiveness of current universal infant rotavirus vaccination strategy,in China.Methods Through constructing decision tree-Markov model,we simulated rotavirus diarrhea associated cost and health outcome on those newborns in 2012 regarding different vaccination programs as:group with no vaccination,Rotavirus vaccination group and Rotateq vaccination group,respectively.We determined the optimal program,based on the comparison between incremental cost-effectiveness ratio (ICER) and China' s 2012 per capital gross domestic product (GDP).Results Compared with non-vaccination group,the Rotavirus vaccination and Rotateq vaccination groups had to pay 3 760 Yuan and 7 578 Yuan (both less than 2012 GDP per capital) to avert one disability adjusted life years (DALY) loss,respectively.Results from sensitivity analysis indicated that both results were robust.Compared with Rotavirus vaccination program,the Rotateq vaccination program had to pay extra 81 068 Yuan (between 1 and 3 times GDP per capital) to avert one DALY loss.Data from the sensitivity analysis indicated that the result was not robust.Conclusion From the perspective of health economics,both two-dose Rotarix vaccine and three-dose's Rotateq vaccine programs were highly cost-effective,when compared to the non-vaccination program.It was appropriate to integrate rotavirus vaccine into the routine immunization program.Considering the large amount of extra cost that had to spend on Rotateq vaccination program,results from the sensitivity analysis showed that it was not robust.Rotateq vaccine required one more dose than the Rotarix vaccine,to be effective.However,it appeared more difficult to practice,suggesting that it was better to choose the Rotarix vaccine,at current stage.

Construction of high altitude military medicine digital library based on Web 3 .0 / 军事医学

Objective To establisha high altitude military medicine digital library ( HAMMDL) in order to integrate high altitude military medicine ( HAMM ) digital resources and promote transformation of HAMM technology to health service application.Methods The solution of Web 3.0-basedHAMMDL was proposed.Ontology, the metadata and Wiki paradigm were used to manage and present resources , while the Model-View-Controller pattern was used in software design .Results Based on analysis of the functional requirements and key techniques of HAMMDL , design principles were proposed the hierarchical software architecture pattern was designed , the software supporting technology was provided and the construction method of HAMM ontologies was discussed .Conclusion With the synthesized application of relative technologies , smart utilization and socialized content management pattern of HAMM digital resources can be achieved .

Prediction of susceptibility to acute mountain sickness based on LVQ neural-network model / 局解手术学杂志

Objective The purpose of this study was to examine the relationship between acute mountain sickness ( AMS) and AMS susceptibility indices before ascent to high altitude and to evaluate their predictive value for AMS. Methods A total of 314 healthy male a-dults were voluntarily enrolled. Their 22 physiological and mental indices of AMS susceptibility were obtained before exposure high altitude. The diagnoses of AMS were based on the Lake Louise score ( LLS) ,an international standard scoring system for AMS. According to the char-acteristics of selected AMS susceptibility indices and the strong fault tolerance of neural network theory, the learning vector quantization ( LVQ) neural network method was adopted to build the prediction model of susceptibility to AMS. Results The results showed the sensitiv-ity of the LVQ model which distinguishes subjects with no-AMS reached 95. 00%,the average correct-prediction precision ultimately reached 72. 22%. The result of prediction is believable. Conclusion The builded LVQ model provide a scientific method for screening crowd who quickly ascend to high altitude,and also can lead to an effective preliminary screening of susceptibility to AMS.

The study of correlation between high-risk types HPV load and expression of nestin in different cervical lesions / 中国医师杂志

Objective To investigate the association between high-risk types of human papillomavirus (HR-HPV) load and the expression of nestin in different cervical lesions.Methods The hybridization capture Ⅱ (HC-Ⅱ) assay was used to test the HR-HPV load from 60 patients who were the first time to do the cervical cancer screening in Xiangya Hospital,and immunohistochemistry was used to detect the expression of nestin in those biopsy tissue samples.Results (1) The lgHPV level (logarithm of HR-HPV load) in the high level lesion group was higher than the low level one (P ＜0.05),and HR-HPV load was positively associated with the degree of cervical lesions (rs =0.269,P =0.037).(2) The expression of nestin in A group was weaker than groups B and C (H =7.271,22.843,P ＜0.01),and the expression of nestin in C group was stronger than B group (H =7.270,P ＜0.01),and the expression of nestin was positively associated with the degree of cervical lesion (rs =0.646,P =0.000).(3) The HR-HPV load was positively associated with the expression of nestin (P ＜ 0.05).Conclusions The HR-HPV load and the expression of nestin are closely related to the cervical lesions,and the joint detection has a referential value for early prevention of cervical lesions and prediction of progress of cervical precancerous lesion.It might guide the early prevention and treatment of cervical cancers.

Design and application research of PE right ventricular catheter for mice / 重庆医学

Objective To establish a simple ,low cost and stable method to detect right ventricular pressure in mice .Methods A PE-50 duct length 15 cm(outside diameter :0 .9 mm ,inner diameter :0 .5 mm) was bent on one terminal and the other terminal was inserted into a 7# syringe needle to connect to a pressure transducer .This duct was intubated into right ventricle via right external jugular vein to detect right ventricular pressure in 80 SPF grade male C57BL/6 mice .Successful cases and operation time were re-corded .Besides ,40 SPF grade male C57BL/6 mice were randomized into the control group (n=20) and chronic hyperbaric hypoxia group(n= 20) .Mice in chronic hyperbaric group were raised in a hyperbaric chamber of simulated 5 000 m high altitude for 4 weeks .The control group was raised outside the chamber simultaneously .Right ventricular systolic pressure was detected with the PE duct .Left and right ventricles were detached and weighed ,and Hermann-Willson index was calculated .Results With this PE duct ,right ventricular intubation success rate was 90% (72/80) ,the operation cost approximately 3 to 5 min each mouse from the separation of blood vessels to detect the time needed for the right ventricle waveform .right ventricular systolic pressure[(39 .52 ± 4 .34 )mm Hg] and Hermann-Willson index(0 .356 ± 0 .039)of chronic hyperbaric hypoxia group were significantly higher than that of control group [(21 .24 ± 2 .7)mm Hg and (0 .256 ± 0 .020)] ,which has significant positive correlation (P<0 .01) .Conclusion It is simple ,fast ,stable ,costing low and of high success ratio to detect right ventricular pressure with this method .

Practice of laboratory animal ethics in medical experiment class and its reflection / 中华医学教育探索杂志

Animal experiment is a primary part of medical education. At present , medical students are lack of education about laboratory animal ethics and they do not usually treat animals fol-lowing rules of laboratory animal ethics. To solve this problem, we focused on the education of labora-tory animal ethics and 3R theory (reduction, replacement, refinement) in order to help the medical students treat laboratory animals nicely. We enforced the construction of the laboratory rules and regu-lations, and improved animal experiments conditions. With these efforts, we successfully standardized and scientized our experiment classes.