Background and purpose:Silent information regulator proteins(sirtuins,SIRT)are a class Ⅲ histone deacetylases with nicotinamide adenine dinucleotide(NAD+)as coenzyme.YME1 like 1 ATPase(YME1L1)is essential for the maintenance of mitochondrial morphology,function and plasticity.Optic atrophy 1(OPA1)mainly mediates mitochondrial fusion.The aim of this study was to explore the expression of SIRT3 in the endocrine resistance of breast cancer,the relationship between SIRT3 and YME1L1 and OPA1,and the mechanism of SIRT3 in the endocrine resistance of breast cancer.Methods:4-hydroxytamoxifen was used to induce tamoxifen-resistant MCF-7/TAM cells.cell counting kit-8(CCK-8)was used to detect cell proliferation and verify drug resistance.The mitochondrial morphology was observed by transmission electron microscopy(TEM)and immunofluorescence staining.The expressions of SIRT3 and OPA1 were detected by real-time fluorescent quantitative polymerase chain reaction(RTFQ-PCR)and Western blot.JC-1 staining was used to detect mitochondrial membrane potential,and dihydroethidium(DHE)staining was used to detect reactive oxygen species(ROS)to verify mitochondrial function.SIRT3 was knocked down in drug-resistant cells by RNA interference,and SIRT3 and YME1L1 wild type(WT),simulated acetylation state mutant(MUT K237Q),and simulated deacetylation state mutant(MUT K237R)were overexpressed in parental cells by overexpression plasmid.Immunoprecipitation assay(IP)and immunofluorescence(IF)were used to verify the interaction between SIRT3 and YME1L1.Results:RTFQ-PCR and Western blot results showed that SIRT3 gene expression and protein level was significantly higher in drug-resistant cells than in parental cells.Overexpression of SIRT3 in parental cells decreased the sensitivity of breast cancer cells to tamoxifen.Knockdown of SIRT3 in drug-resistant cells enhanced the sensitivity of drug-resistant cells to tamoxifen.DHE staining showed that the ROS level was lower in tamoxifen resistant cells than in parental cells at the same concentration.Transmission electron microscopy and fluorescence staining showed that the mitochondria of the drug-resistant cells were elongated compared with the parental cells.Western blot results showed that the expression level of L-OPA1 protein was higher in drug-resistant cells than in parental cells.Overexpression of SIRT3 in the parental cells resulted in enhanced mitochondrial function and longer mitochondrial morphology compared with the control cells.Western blot showed that the expression of L-OPA1 was upregulated.When SIRT3 was knocked down in drug-resistant cells,the opposite result was obtained.We further verified how SIRT3 regulated OPA1 protein,affected the morphology and function of mitochondria,and promoted drug resistance of breast cancer.Overexpression of YME1L1(wild-type and mutant plasmids)in parental cells showed that overexpression of YME1L1 in the simulated deacetylation state resulted in similar results as overexpression of SIRT3,and overexpression of YME1L1 in the acetylated state resulted in similar results as knockdown of SIRT3.IP assay confirmed the interaction between SIRT3 and YME1L1 in breast cancer cells.The acetylation level of YME1L1 was different at different SIRT3 expression levels.IF assay showed that YME1L1 was co-localized with SIRT3 in MCF-7 cells.Conclusion:SIRT3 is highly expressed in tamoxifen-resistant breast cancer cells.SIRT3 upregulates L-OPA1 expression by deacetylating YME1L1,thereby promoting mitochondrial fusion and enhancing mitochondrial function,and promotes tamoxifen resistance in breast cancer.

Objective:To investigate the feasibility of conducting active surveillance (AS) for low risk papillary thyroid microcarcinoma (PTMC) in China and to examine the factors in association with disease progression during AS.Methods:This study was a prospective observational research conducted from Jun. 2018 to Aug. 2022 at Ruijin Hospital affiliated to Shanghai Jiao Tong University School of Medicine. Seventy-three patients with cytologically confirmed low-risk PTMC were enrolled in this study. They were followed up by ultrasonography, and the observed nodules were re-assessed whether or not to have disease progression. Disease progression was defined as having nodule enlarged more than 3 mm in any of diameters measured on ultrasound, or/and presence of suspicious lymph node metastasis.Results:The median follow-up time was 33 months. At the time of last follow-up, 16 cases (21.9%) exhibited disease progression, including 9 cases (12.3%) with suspicious lymph nodes detected by ultrasound, and 8 cases (11.0%) with lesion enlargement; one case (1.3%) exhibited both situations. The univariate chi-square analysis revealed that young patients (≤45 years old, P=0.041), presence of microcalcifications ( P=0.032), initial larger nodule (diameter greater than 7 mm, P=0.003), and elevated thyroid autoantibody levels ( P=0.008) were associated with disease progression. Multiple regression analysis showed elevated thyroid autoantibodies ( OR=4.311, P=0.030) and initial larger nodule ( OR=6.196, P=0.034) were independent risk factors for PTMC progression,respectively. Conclusions:AS for low-risk PTMC is a feasible and effective. During the observation, ultrasound can reveal suspicious lymph nodes and nodule enlargement, which are crucial indicators for assessing disease progression. Patients with initially larger nodule size and elevated thyroid autoantibody level are more likely to exhibit disease progression and should receive closer attention.

Objective To establish the fingerprint map and method of multi-component content determination of ginger and its processed products,and to evaluate their quality. Methods High performance liquid chromatography (HPLC) was used to establish the fingerprint of ginger and its processed products (dried ginger,baked ginger,ginger charcoal) and the method for the simultaneous determination of content of five components. The data were analyzed by similarity evaluation and chemical pattern recognition. Results The fingerprint of ginger and its processed products was established. The similarity was ranged from 0.931 to 0.996. A total of 15 common peaks were confirmed. Five components (6-ginger phenol,8-ginger phenol,10-ginger phenol,6-ginger enol,zingiberone) were identified when compared with the standard. The content of 6-ginger phenol,8-ginger phenol and 10-ginger phenol in ginger and its processed products(dried ginger,baked ginger,ginger charcoal),which were determined by HPLC,met the standard of Chinese Pharmacopoeia(2020 edition). The content of 6-ginger enol and zingiberone in fresh ginger was very low,but their content increased significantly after processing into dried ginger and baked ginger. Ginger and its processed products can be divided into different category in cluster analysis. Five components with VIP greater than one were selected by orthogonal partial least squares discriminant analysis. Conclusion The HPLC fingerprint and assay method established in this study are specific,simple and feasible,stable and reliable. With the help of chemical pattern recognition method,it can provide reference for the quality evaluation of ginger and its processed products.

OBJECTIVE: To explore the clinical effect of Li-Dan-He-Ji in the treatment of infantile cholestatic hepatic fibrosis. METHODS: Patients who met the diagnostic criteria of infantile cholestatic hepatic fibrosis in the department of integrated traditional Chinese and Western medicine and the department of gastroenterology of Wuhan Children's Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from January to December 2021 were included in the study by prospective randomized controlled trial. They were divided into the conventional treatment group and Li-Dan-He-Ji group according to the random number table. The patients in the conventional treatment group were given conventional treatment according to the guidelines. In the Li-Dan-He-Ji group, the self-made Chinese medicinal compound Li-Dan-He-Ji (prescription: Herba Artemisiae Scopariae, Fructus Forsythiae, Radix et Rhizoma Rhei preparata, Radix Polygoni Multiflori Preparata, Radix Paeoniae Rubra, Ramulus Cinnamomi, Fructus Aurantii, Rhizoma Atractylodis Macrocephalae, Fructus Schisandrae Chinensis, Carapax Trionycis, and Radix Glycyrrhizae) was given on the basis of the routine treatment, by oral, enema or nasal feeding, 60 mL each day, divided into 2 or 3 times, for 28 days. Outpatient follow-up was maintained for 4 weeks. Before and after treatment, serum liver fibrosis 4 items [type IV collagen (IV-C), hyaluronidase (HA), type III procollagen (PC III), laminin (LN)], liver function and cholestasis-related markers [total bilirubin (TBil), direct bilirubin (DBil), total bile acid (TBA), alkaline phosphatase (ALP), γ-glutamyl transpeptidase (γ-GGT), alanine aminotransferase (ALT), aspartate aminotransferase (AST)], oxidative stress markers [superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH)], liver stiffness measurement (LSM) detected by transient elastography (TE), aspartate aminotransferase-to-platelet ratio index (APRI), and liver and spleen retraction time were recorded in the two groups. RESULTS: During the observation period, a total of 40 cases of cholestatic hepatic fibrosis were treated, including 21 cases in the conventional treatment group and 19 cases in the Li-Dan-He-Ji group. Before treatment, the differences in serum liver fibrosis 4 items, serum liver function and cholestasis-related markers, oxidative stress indexes, LSM and APRI of the two groups were not statistically significant. After treatment, the liver fibrosis 4 items, liver function and cholestasis-related markers, LSM, and APRI were all significantly decreased in both groups, and the indexes in the Li-Dan-He-Ji group were significantly lower than those in the conventional treatment group [HA (ng/L): 165.81±21.57 vs. 203.87±25.88, PC III (μg/L): 69.86±9.32 vs. 81.82±7.39, IV-C (μg/L): 204.14±38.97 vs. 239.08±24.93, LN (μg/L): 162.40±17.39 vs. 190.86±15.97, TBil (μmol/L): 37.58±27.63 vs. 53.06±45.09, DBil (μmol/L): 20.55±19.34 vs. 30.08±27.39, ALP (U/L): 436.50±217.58 vs. 469.60±291.69, γ-GGT (U/L): 66.78±35.84 vs. 87.00±32.82, ALT (U/L): 64.75±50.53 vs. 75.20±50.19, AST (U/L): 77.25±54.23 vs. 96.80±59.77, TBA (μmol/L): 74.35±44.44 vs. 85.45±39.50, LSM (kPa): 5.24±0.39 vs. 7.53±3.16, APRI: 0.52±0.39 vs. 0.98±0.29, all P < 0.05]. After treatment, MDA in the two groups were significantly lower than those before treatment, and SOD and GSH were significantly higher than those before treatment. The level of SOD in the Li-Dan-He-Ji group was significantly higher than that in the conventional treatment group (kU/L: 64.56±6.69 vs. 51.58±5.98, P < 0.05). In addition, the liver retraction time (day: 20.13±10.97 vs. 24.33±13.46) and spleen retraction time (day: 25.93±13.01 vs. 29.14±14.52) in the Li-Dan-He-Ji group were significantly shorter than those in the conventional treatment group (both P < 0.05). CONCLUSIONS The use of Li-Dan-He-Ji in the treatment of cholestatic hepatic fibrosis can effectively improve the indicators of cholestasis, hepatic fibrosis, oxidative stress and clinical symptoms in children.
Child ; Humans ; Prospective Studies ; Cholestasis/pathology* ; Liver ; Liver Cirrhosis/drug therapy* ; Bilirubin/pharmacology* ; Oxidative Stress ; Aspartate Aminotransferases/metabolism* ; Superoxide Dismutase/metabolism*

Objective:To observe any effect of electroacupuncture (EA) on the expression of phosphorylated extracellular signal-regulated protein kinase (p-ERK1/2) and phosphorylated cyclic adenosine monophosphate response element binding protein (p-CREB) in the spinal dorsal horns of diabetics experiencing neuropathic pain.Methods:Eight rats were randomly selected from 30 healthy male Sprague-Dawley rats as the normal group (N), and the remaining twenty-two rats were treated with a single high-dose intraperitoneal injection of streptozotocin (STZ) to establish a neuropathic pain model. The rats modeled successfully were randomly divided into a model group (M, n=8) and an EA group ( n=8). In the EA group, electroacupuncture was applied at the bilateral Hou san li and Kunlun acupoints starting on the 15th day after the STZ injection. The daily sessions lasted 30 minutes for 1 week. Body weight (BW), fasting blood glucose (FBG) and paw withdrawal latency (PWL) were observed before the STZ injection and on the 7th, 14th, and 21st days afterward. The expression of p-ERK1/2 and p-CREB in the dorsal horns of the rats′ spinal cords was detected using western blotting. The count of p-CREB-positive cells in the dorsal horns and their co-localization with neurons was detected using immunofluorescence. Results:In comparison with the N group, the average BW of the M group on the 7th, 14th and 21st days after the STZ injection was significantly lower, while the average FBG was significantly higher. There was no significant difference between the M and N groups in the average PWL on the 7th day after the STZ injection, but it had decreased significantly in the M group on the 14th and 21st days. Compared with the M group, the average PWL of the EA group was significantly longer on the 21st day after the injection. The expression of p-ERK1/2 and p-CREB protein in the spines of the M group was significantly higher than in the N group. p-CREB positive cells were more numerous in the M group compared with the N group, while in the EA group they were fewer. P-CREB was co-located with neurons in the spinal dorsal horn.Conclusion:EA can alleviate neuropathic pain effectively, perhaps by inhibiting the expression of p-ERK1/2 and p-CREB in the dorsal horns of the spinal cord.

Objective:To investigate the effect of huaier granule combined with docetaxel + epirubicin + cyclophosphamide (TEC) neoadjuvant chemotherapy on the disease control rate and the levels of extracellular fragment of human epidermal growth factor receptor 2 (HER2-ECD) and tumor abnormal protein (TAP) in patients with advanced breast cancer.Methods:From April 2013 to June 2016, 94 patients with advanced breast cancer in Shengjing Hospital Affiliated to China Medical University were selected and randomly divided into observation group ( n=47) and control group ( n=47). The control group was treated with TEC neoadjuvant chemotherapy, and the observation group was treated with TEC neoadjuvant chemotherapy combined with huaier granules for 12 weeks. The curative effect, incidence of adverse reactions, cellular immune function indexes (CD8 + , CD4 + , CD4 + /CD8 + ), serum HER2-ECD and TAP levels were compared before and 12 weeks after treatment. The patients were followed up for 3 years after treatment, and the 1-year, 2-year and 3-year survival rates were compared between the two groups. Results:After 12 weeks of treatment, the disease control rate of the observation group was higher than that of the control group, with statistically significant difference ( P<0.05); during the treatment, the incidence of gastrointestinal reaction, neutropenia, thrombocytopenia and abnormal liver function of the observation group was lower than that of the control group, with statistically significant difference ( P<0.05); after 12 weeks of treatment, the serum CD8 + level of the observation group was lower than that of the control group, and the serum CD4 + , CD4 + /CD8 + level was higher than that of the control group, with statistically significant difference ( P<0.05); after 12 weeks of treatment, the serum levels of HER2-ECD and TAP in the observation group were lower than those in the control group, with statistically significant difference ( P<0.05); the 3-year survival rate in the observation group was higher than that in the control group, with statistically significant difference ( P<0.05). Conclusions:Huaier granules combined with TEC neoadjuvant chemotherapy in patients with advanced breast cancer can reduce serum HER2-ECD, TAP levels, which can enhance the immune function of the body, reduce the toxic and side effects, improve the long-term survival rate, and have a significant effect.

Objective:To develop an R script that can efficiently and accurately filter genome-wide association studies (GWASs) from the GWAS Catalog Website.Methods:The selection principles of GWASs were established based on previous studies. The process of manual filtering in the GWAS Catalog was abstracted as standard algorithms. The R script (gwasfilter.R) was written by two programmers and tested many times.Results:It takes six steps for gwasfilter.R to filter GWASs. There are five main self-defined functions among this R script. GWASs can be filtered based on "whether the GWAS has been replicated" "sample size" "ethnicity of the study population" and other conditions. It takes no more than 1 second for this script to filter GWASs of a single trait.Conclusions:This R script (gwasfilter.R) is user-friendly and provides an efficient and standard process to filter GWASs flexibly. The source code is available at github ( https://github.com/lab319/gwas_filter).

OBJECTIVE： To evaluate the efficacy and safety of apremilast in the treatment of moderate-to-severe plaque psoriasis systematically. METHODS： Retrieved from PubMed， Embase， Cochrane Library， VIP， CNKI and CBM， RCTs about apremilast or apremilast combined with other drugs （trial group） versus placebo （control group） in the treatment of moderate- to-severe plaque psoriasis were collected. Meta-analysis was conducted by using Rev Man 5.3 statistical software after literature screening， data extraction and quality evaluation with bias risk evaluation tool of Cochrane System Evaluator Manual 5.1.0. RESULTS： Totally 7 studies were included， involving 2 332 patients. Results of Meta-analysis showed that case number of psoriasis assessment and severity index （PASI） decreased by 75％ （PASI 75％） [OR＝6.44，95％CI（4.90，8.45），P＜0.000 01]， PASI 90％ [OR＝8.13， 95％CI（4.65， 14.22）， P＜0.000 01] and sPGA 0 or 1 [OR＝3.89，95％CI（3.00，5.05），P＜0.000 01]， the incidence of ADR [OR＝1.87，95％CI（1.44，2.43）， P＜0.000 01] in trial group were significantly more or higher than control group. Subgroup analysis by apremilast dose showed that case number of 20 mg PASI 75％ [OR＝4.72，95％CI（2.77，8.05），P＜0.000 01]， 30 mg PASI 75％ [OR＝7.05，95％CI（5.13，9.69），P＜0.000 01]， 20 mg PASI 90％ [OR＝4.27，95％CI（1.80，10.09），P＝0.001]， 30 mg PASI 90％ [OR＝11.11，95％CI（5.27，23.43），P＜0.000 01]， 20 mg sPGA 0 or 1 [OR＝2.82，95％CI（1.51，5.26），P＝0.001]， 30 mg sPGA 0 or 1 [OR＝4.13，95％CI（3.10，5.50），P＜0.000 01]， the incidence of 30 mg ADR [OR＝1.94，95％CI（1.51，2.49），P＜0.000 01] in trial group were significantly more or higher than control group. There was no statistical significance in the incidence of serious ADR [OR＝1.27，95％CI（0.77，2.07），P＝0.35] or case number of ADR leading to withdrawal [OR＝1.48，95％CI（1.00，2.20），P＝0.05] between 2 groups. CONCLUSIONS： Apremilast is effective for moderate-to-severe plaque psoriasis in dose-dependent manner and improve the quality of life， but increase the incidence of ADR.

Objective To investigate the method of modeling, finite element modeling and AnyBody musculoskeletal multi-body dynamics simulation technique analyze the biomechanics of clinical orthopaedic surgery. Methods The AnyBody software was used to establish the musculoskeletal motor model of the individualized upper limbs according to the height, weight and CT data of the volunteers. The flexion motion of the elbow in normal people was simulated, and the muscle force, joint force, torque, constraint condition of the humerus during the flexion movement were derived and used as the boundary conditions of finite element analysis.Then, the 3D reconstruction was conducted in the MIMICS software based on CT data. In the Geomagic Studio software, the humeral curved surface and position coordinate matching were completed, and grid division and material assignment were done in the HyperMesh software. Finally, the 3D reconstruction for finite element model of the humerus was introduced into ABAQUS software. The boundary condition data derived from the AnyBody software were applied and the stress calculation analysis was performed. Results The results of the stress and displacement of the humerus during elbow flexion motion were calculated in the ABAQUS software. The maximum stress and displacement of the humerus were 0.76 MPa and 20 μm when flexion of the elbow joint was about 90°. Conclusions A continuous dynamic analysis of humeral stress and displacement during elbow flexion motion was realized, which was more consistent with the requirements of human physiological anatomy and could provide an efficient analysis platform and a new way for studying clinical orthopedic problems.

Objective To examine the feasibility and early effects of unilateral pedicle screw fixation in combination with contralateral translaminar facet screw fixation plus interbody fusion through the channel-assisted muscle-splitting approach for the treatment of degenerative lumbar spine diseases in elderly patients.Methods A total of 82 elderly patients with degenerative lumbar spine diseases received treatment between January 2015 and June 2016.Of these patients,25 had lumbar spinal stenosis,42 had degenerative lumbar instability,13 had giant lumbar disc herniation,and two had recurrent lumbar disc herniation;There were 69 cases of single segment and 13 cases of double segment lesions.Perioperative evaluation,imaging evaluation,and clinical evaluation were performed,and surgical complications were analyzed.Results The average length of incision was (2.6 ± 0.4) cm in cases of single segment lesions and (4.4±0.5) cm in cases of two segment lesions.The mean operation time was (98.3±19.6) minutes and peri-operation blood loss was (232.2±25.8) ml.The post-operation height of the intervertebral disc space significantly increased at the last follow-up(P ＜ 0.05);the height of the intervertebral disc space was restored and well maintained with internal fixation showing no loosening,breakage or Cage shift.The post-operation lumbar coronal Cobb angle decreased(P＜0.05)while the sagittal Cobb angle increased(P＜0.05)at the last follow-up.There was no significant difference in the area and grade of the multifidus muscle on MRI 12 months after operation.Seventy-two patients were followed up for 12-18 months (mean =16.8 months),and there were 67 cases of intervertebral fusion (93.1％).The last follow-up showed a Japanese Orthopaedic Association(JOA) score of (25.8 ± 1.7),which was significantly higher than the pre-operation score (10.6±1.3),with an excellence rate of 88.9 ％.The Oswestry Disability Index score was significantly reduced,from (50.9±21.6) at pre-operation to (9.6± 4.8) at the last follow-up.There were 78 patients with primary wound healing,three cases of superficial wound necrosis,one case of dural tear,and one case of nerve root injury.No cardiovascular or cerebrovascular incident was observed during or after operation.Conclusions Combined pedicle/facet screw fixation plus interbody fusion through the channel-assisted muscle-splitting approach shows satisfactory short-term effects when used for the treatment of degenerative lumbar spine diseases in the elderly and possesses advantages such as minimal invasiveness,limited bleeding,quick recovery,high fusion rates and significantly improved clinical outcomes.However,further clinical studies are required to assess its medium-and long-term efficacy.