ObjectiveThe proteome of biological evidence contains rich genetic information, namely single amino acid polymorphisms (SAPs) in protein sequences. However, due to the lack of efficient and convenient analysis tools, the application of SAP in public security still faces many challenges. This paper aims to meet the application requirements of SAP analysis for forensic biological evidence’s proteome data. MethodsThe software is divided into three modules. First, based on a built-in database of common non-synonymous single nucleotide polymorphisms (nsSNPs) and SAPs in East Asian populations, the software integrates and annotates newly identified exonic nsSNPs as SAPs, thereby constructing a customized SAP protein sequence database. It then utilizes a pre-installed search engine—either pFind or MaxQuant—to perform analysis and output SAP typing results, identifying both reference and variant types, along with their corresponding imputed nsSNPs. Finally, SAPTyper compares the proteome-based typing results with the individual’s exome-derived nsSNP profile and outputs the comparison report. ResultsSAPTyper accepts proteomic DDA mass spectrometry raw data (DDA acquisition mode) and exome sequencing results of nsSNPs as input and outputs the report of SAPs result. The pFind and Maxquant search engines were used to test the proteome data of 2 hair shafts of2 individuals, and both obtained SAP results. It was found that the results of the Maxquant search engine were slightly less than those of pFind. This result shows that SAPTyper can achieve SAP fingding function. Moreover, the pFind search engine was used to test the proteome data of 3 hair shafts from 1 European person and 1 African person in the literature. Among the sites fully matched by the literature method, sites detected by SAPTyper are also included; for semi-matching sites, that is, nsSNPs are heterozygous, both literature method and SAPTyper method had the risk of missing detection for one type of the allele. Comparing the analysis results of SAPTyper with the SAP test results reported in the literature, it was found that some imputed nsSNP sites identified by the literature method but not detected by SAPTyper had a MAF of less than 0.1% in East Asian populations, and therefore they were not included in the common nsSNP database of East Asian populations constructed by this software. Since the database construction of this software is based on the genetic variation information of East Asian populations, it is currently unable to effectively identify representative unique common variation sites in European or African populations, but it can still identify SAP sites shared by these populations and East Asian populations. ConclusionAn automated SAP analysis algorithm was developed for East Asian populations, and the software named SAPTyper was developed. This software provides a convenient and efficient analysis tool for the research and application of forensic proteomic SAP and has important application prospects in individual identification and phenotypic inference based on SAP.

OBJECTIVE To study the mechanism of oxymatrine inducing apoptosis of osteosarcoma MG63 cell line based on mitophagy mediated by cyclooxygenase-2 （COX-2）/PTEN-induced putative kinase-1 （PINK1）/Parkinson disease protein-2 （Parkin） signaling pathway. METHODS MG63 cells were treated with 2.0， 4.0， 8.0 mg/mL oxymatrine and 6 μmol/L 5-fluorouracil， then the apoptotic rate， the expression of apoptosis-related proteins [B-cell lymphoma-2 （Bcl-2）， Bcl-2 related X protein （Bax）]， the proportion of decrease in mitochondrial membrane potential， the level of mitophagy as well as the protein expressions of PINK1， Parkin， and microtubule-associated protein 1 light chain-3Ⅱ （LC3-Ⅱ） were detected. PINK1 small interfering RNA （siRNA） was adopted to intervene in the expression of PINK1， the cells were divided into control group， PINK1 siRNA group， oxymatrine group， and PINK1 siRNA+oxymatrine group； the protein expressions of PINK1， Parkin， and LC3-Ⅱ， the proportion of decrease in mitochondrial membrane potential （MMP） as well as apoptotic rate were detected. The lentivirus infection technique was used to overexpress COX-2， the cells were divided into control group， oxymatrine group， COX-2 group， and COX-2+oxymatrine group. The protein expressions of COX-2， PINK1， and Parkin， as well as the proportion of decrease in MMP were detected. RESULTS After being treated with oxymatrine， the apoptotic rate， the protein expressions of Bax， PINK1， Parkin， and LC3-Ⅱ， the level of mitophagy as well as the proportion of decrease in MMP were significantly increased （P＜0.05）， while the protein expression of Bcl-2 was significantly decreased （P＜0.05）. Compared with the oxymatrine group， the protein expressions of PINK1， Parkin， and LC3-Ⅱ， apoptotic rate and the proportion of decrease in MMP were significantly decreased in PINK1 siRNA+oxymatrine group （P＜0.05）. Compared with the oxymatrine group， the protein expression of COX-2 in the COX-2+oxymatrine group was increased significantly （P＜0.05）， while the protein expressions of PINK1 and Parkin as well as the proportion of 526087266@qq.com decrease in MMP were decreased significantly （P＜0.05）. CONCLUSIONS Oxymatrine can mediate the overactivity of mitophagy based on the PINK1/Parkin signaling pathway by inhibiting COX-2 expression， thus promoting the apoptosis of the MG63 osteosarcoma cell line.

OBJECTIVE To investigate the effect of berberine on ferroptosis in MG63 osteosarcoma cells and its mechanism. METHODS Using cells without drug treatment as control， the cell viability， proliferation， the related indexes of ferroptosis [nuclear proliferation associated-antigen （Ki67）， mitochondrial ultrastructure， ferric ion （Fe2+）， reactive oxygen species （ROS）， malondialdehyde （MDA）， and glutathione （GSH）]， the protein expression of signal transducer and activator of transcription 3 （STAT3）， tumor protein 53 （p53）， and solute carrier family 7 member 11 （SLC7A11） were detected after being treated with different concentrations of berberine. Cells were transfected with p53 siRNA and then assigned to the control group， p53 siRNA group， berberine group， and p53 siRNA+berberine group to explore the role of p53 in berberine-induced ferroptosis. After 24 h incubation with 10.0 μmol/L berberine， the protein expressions of p53 and SLC7A11， the levels of mitochondrial membrane potential， GSH， and MDA content were determined. Cells were transfected with STAT3 overexpressed plasmid and then assigned to the control group， berberine group， STAT3 group， and STAT3+berberine group to explore the effect of STAT3 on the regulation of the p53/SLC7A11 pathway. After 24 h incubation with 10 μmol/L berberine， the protein expressions of p-STAT3， STAT3， p53， and SLC7A11 were detected. RESULTS Compared with the control cell， the concentrations of 2.5， 5.0 and 10.0 μmol/L berberine could reduce the cell viability and expression of Ki67， and induce the morphological changes in ferroptosis-related mitochondria， increase the levels of Fe2+， ROS and MDA， and the protein expression of p53， reduce the level of GSH， the binding activity of STAT3 with DNA， and the protein expressions of p-STAT3 and SLC7A11； the above differences were statistically significant （P＜ 0.05 or P＜0.01）. Compared with the berberine group，significantly down-regulated p53 protein expression and MDA level， up-regulated SLC7A11 protein expression， and increased mitochondrial membrane potential and GSH level were observed in the p53 siRNA+berberine group （P＜0.01）. Compared with the berberine group， the protein expressions of p-STAT3， STAT3， and SLC7A11 in the STAT3+berberine group were significantly increased （P＜0.01）， while the protein expression of p53 was significantly decreased （P＜0.01）. CONCLUSIONS Berberine can induce the ferroptosis of MG63 cells by mediating STAT3/p53/SLC7A11 signaling pathway.

Chronic urticaria （CU） is a common skin disease worldwide， and its incidence is increasing year by year in various regions. Clinical manifestations such as severe itching can affect normal work， sleep， and daily life and increase the negative psychological burden caused by stress， anxiety， and depression. Mast cell activation and degranulation induced by immunoglobulin（Ig）E hypersensitivity is one of the core pathogenic mechanisms of CU， and there is no cure. Antihistamines such as cetirizine and loratadine are preferred for the clinical treatment of CU. Although they can effectively improve clinical manifestations such as itchiness， long-term application can increase the risk of adverse reactions and drug resistance. The phosphatidylinositol kinase/serine-threonine protein kinase B（PI3K/Akt） signaling pathway， as a classical signaling pathway regulated by phosphatidylinositol and tyrosine kinase receptor （RTK）， is a key target regulating the production and release of cytokines in macrophages and affecting the migration of leukocytes and the activation of mast cells and inflammation， and it can be involved in a variety of metabolic processes， such as mast cell activation and degranulation induced by IgE hypersensitivity and abnormal activation of the complement system so that the PI3K/Akt molecular pathway could be an important target for the future eradication of CU. However， the mechanism and potential role of the PI3K/Akt signaling pathway in the treatment of CU are less reported in China. Now， this paper reviewed the molecular mechanism of PI3K/Akt signaling pathway regulation in the treatment of CU and provided corroborative evidence and therapeutic strategy choices for the treatment of CU with traditional Chinese medicine （TCM） from the perspectives of molecular regulation and network pharmacology analysis.

Tuberous sclerosis complex(TSC)is a rare genetic disease that can lead to benign dysplasia in multiple organs such as the skin, brain, eyes, oral cavity, heart, lungs, kidneys, liver, and bones. Its main symptoms include epilepsy, intellectual disabilities, skin depigmentation, and facial angiofibromas, whilst incidence is approximately 1 in 10 000 to 1 in 6000 newborns. This case presents a middle-aged woman who initially manifested with epilepsy and nodular depigmentation. Later, she developed a lower abdominal mass, elevated creatinine, and severe anemia. Based on clinical features and whole exome sequencing, the primary diagnosis was confirmed as TSC. Laboratory and imaging examinations revealed that the lower abdominal mass originated from the uterus. CT-guided biopsy pathology and surgical pathology suggested a combination of leiomyoma and abscess. With the involvement of multiple organs and various complications beyond the main diagnosis, the diagnostic and therapeutic process for this patient highlights the importance of rigorous clinical thinking and multidisciplinary collaboration in the diagnosis and treatment of rare and challenging diseases.

Zuoguiyin， which first recorded in Jingyue Quanshu written by ZHANG Jiebin in the Ming dynasty， was included in the Catalogue of Ancient Famous Classical Formulas（The Second Batch）. This study followed the Principles of Textual Research on Key Information of Ancient Famous Classical Formulas to determine the key information of Zuoguiyin in ancient and modern literature， such as the formula origin， the composition of the formula and the origin of the drugs. It was found that the composition， dosage， preparation and processing methods of Zuoguiyin were basically the same as the original formula. The original dosage of this formula is 41.03 g of Rehmanniae Radix Praeparata（the fresh or dried tuberous roots of Rehmannia glutinosa， processed by wine stewing or wine steaming）， 7.46 g of Dioscoreae Rhizoma（the dried rhizomes of Dioscorea opposita）， 7.46 g of Lycii Fructus（the dried mature fruit of Lycium barbarum）， 3.73 g of Glycyrrhizae Radix et Rhizoma Praeparata cum Melle（the dried roots and rhizomes of Glycyrrhiza uralensis， processed by honey-roasted method）， 5.595 g of Poria（the sclerotium of Poria cocos）， 5.595 g of Corni Fructus（the dried mature fruit pulp of Cornus officinalis）. The method of administration is to add 600 mL of water to all the herbs， decoct to 140 mL and take before meals. The function of Zuoguiyin is to nourish Yin and tonify the kidney， and it is often used in the treatment of lumbar soreness and ejaculation， night sweating， dry mouth and throat， thirst and desire to drink， glossy red tongue， thin and rapid pulse， etc. Since ancient times， Zuoguiyin has been used to treat a variety of internal and gynaecological diseases as well as diseases of the nervous， circulatory and reproductive systems that are predominantly caused by kidney Yin deficiency. However， there is not much research on the modern application and therapeutic mechanism of this formula， and there is no standardized preparation in the market， so the degree of development and utilization is not high， and there is still a lot of room for research.

Tetraacetyl phytosphingosine (TAPS) is an excellent raw material for natural skin care products. Its deacetylation leads to the production of phytosphingosine, which can be further used for synthesizing the moisturizing skin care product ceramide. For this reason, TAPS is widely used in the skin care oriented cosmetics industry. The unconventional yeast Wickerhamomyces ciferrii is the only known microorganism that can naturally secrete TAPS, and it has become the host for the industrial production of TAPS. This review firstly introduces the discovery, functions of TAPS, and the metabolic pathway for TAPS biosynthesis is further introduced. Subsequently, the strategies for increasing the TAPS yield of W. ciferrii, including haploid screening, mutagenesis breeding and metabolic engineering, are summarized. In addition, the prospects of TAPS biomanufacturing by W. ciferrii are discussed in light of the current progresses, challenges, and trends in this field. Finally, guidelines for engineering W. ciferrii cell factory using synthetic biology tools for TAPS production are also presented.
Sphingosine ; Ceramides ; Metabolic Engineering ; Synthetic Biology

Objective To investigate the contamination of four polycyclic aromatic hydrocarbons (PAHs) in domestic pre-packaged edible vegetable oil sold in Henan Province. Methods A total of 212 domestic edible vegetable oil samples from Henan Province were collected. The contents of four polycyclic aromatic hydrocarbons (benzo[a]anthracene, chrysene, benzo[b]fluoranthene, and benzo[a]pyrene, defined as PAH4) were determined by HPLC and gas chromatography-mass spectrometry in 2019 and 2020, respectively. Results The content of PAH4 in domestic pre-packaged edible vegetable oils ranged from 0.34 μg/kg to 26.6 μg/kg, the detection rate was 83.49%, and the over-standard rate was 3.30% according to EU standard. In terms of food category, the peanut oil samples were most seriously contaminated with PAH4. From the point of view of product grade and production process, with the decrease of sample quality index, the risk of PAH4 contamination in sesame oil increased. According to GB 2762-2017, the content of benzo (a) pyrene in all samples was qualified, while the peanut oil had the highest rate exceeding the EU standard, with the over-standard rate of 38.46%. Peanut oil had a higher risk of being contaminated by benzo [a] pyrene. Conclusion Peanut oil in domestic pre-packaged edible vegetable oil sold in Henan Province has a high risk of PAH4 contamination. With the decrease of sesame oil quality index, the risk of PAH4 contamination increases.

Objective:To observe the efficacy of modified Simotang in treatment of adult functional constipation (Qi-stagnancy constipation), and investigate its effects on serum levels of intestinal neurotransmitter nitric oxide synthase (nNOS), nitric oxide (NO), and vasoactive intestinal peptide (VIP), as well as superoxide dismutase (SOD), malonaldehyde (MDA), and glutathione (GSH) levels. Method:One hundred and ten patients with functional constipation were selected and randomly divided into control group (55 cases) and treatment group (55 cases) by referring to random number table. The patients in control group were given with routine therapy, Domperidone tablet (1 tablet/time, tid), and Phenolphthalein tablets (100 mg/time, bid). The patients in treatment group were treated with modified Simotang, 1 dose/day. Both groups were treated for 4 weeks. Then the scores of main clinical symptoms of functional constipation, scores of Qi-stagnancy constipation and clinical efficacy were compared between two groups. Constipation recurrence rate was compared between two groups after stopping medicine. Serum levels of intestinal neurotransmitters nNOS, NO and VIP as well as SOD, MDA, GSH levels were detected in both groups. Result:After treatment, scores for main clinical symptoms (difficult defecation, abdominal distension, defecation time, number of defecation times) and Bristol scores in treatment group were obviously lower than those in control group (P<0.01). Scores for symptoms of Qi-stagnancy constipation (ungratifying defecation, abdominal distension,bowel ringing, frequent flatus, chest and flank tightness) in treatment group were obviously lower than those in control group after treatment (P<0.01). Total clinical efficacy in treatment group 98.04% was superior to that in control group 84.62% ( P<0.05). Constipation recurrence rate was 3.92% and 8.16% in treatment group after 4 and 8 weeks of medication stopping, obviously lower than those in control group 18.18% and 27.78% (P<0.05). After treatment, serum levels of intestinal neurotransmitters nNOS, NO, VIP in treatment group were remarkably lower than those in control group (P<0.01). After treatment, serum levels of SOD and GSH in treatment group were higher while MDA level was lower than those in control group (P<0.01). Conclusion:Based on routine therapy, modified Simotang in treatment of adult functional constipation (Qi-stagnancy constipation) can improve clinical symptoms and syndrome symptoms, increase the clinical efficacy, decrease recurrence rate, and regulate levels of intestinal neurotransmitters nNOS,NO and VIP as well as SOD, MDA, GSH levels.

OBJECTIVETo explore the anti-myeloma effect of suberoylanilide hydroxamic acid (SAHA) and on mouse myeloma cell line SP2/0 in vitro and in vivo and its mechanism.

METHODSThe inhibitory effect of SAHA on SP2/0 cells was measured by CCK-8 assay,and the apoptosis and cell cycle were analyzed by flow cytometry FACS. The protein expression of Caspase-3 and p53 of SP2/0 cells treated with SAHA were examined by Western blot. Annexin V/7-AAD double staining was performed to detect the apoptosis of SP2/0 induced by SAHA in vitro. SP2/0 cells (1×10) resuspended in 200 µl PBS were inoculated subcutaneously and intravenously into BALB/c mice, so as to establish aggressive or non-aggressive myeloma-bearing mouse models respectively. On day 3 after modeling, mice received SAHA or vehicle control treatment by intraperitoneal injection. The dose of SAHA was 60 mg/kg·d, 5 times a week for 3 weeks.

RESULTSIn SAHA-treated SP2/0 cells, the proliferation inhibition rate and apoptotic cells increased in a dose dependent manner. Also, SAHA significantly increased the ratio of cells in G phase and decreased in S phase. Molecular mechanisms of apoptosis and cell cycle arrest of SP2/0 induced by SAHA partly correlated with up-regulating the expression level of Caspase-3 and p53. In the non-aggressive myeloma-bearing mice, SP2/0 cells disappeared in peripheral blood after SAHA treatment. In the aggressive myeloma-bearing mice, inhibition of tumor growth and prolongation of the cell survival were observed after SAHA treatment.

CONCLUSIONSAHA inhibited SP2/0 cell proliferation, this effect associates with inducing apoptosis and cell cycle arrest, the mechanism of SAHA ralates partly with activating Caspase-3 and p53 pathway.

Animals ; Antineoplastic Agents ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Histone Deacetylase Inhibitors ; Hydroxamic Acids ; Mice ; Mice, Inbred BALB C ; Multiple Myeloma