Perilla frutescens (L.) Britt. is a characteristic oil crop rich in polyunsaturated fatty acids, particularly α-linolenic acid, which has important development and utilization value. The Dof transcription factor is one of the plant-specific transcription factor families, which is widely involved in important biological processes such as plant growth, development, and metabolic regulation. In order to explore the key Dof transcription factors involved in the oil biosynthesis and systematically analyze their regulatory mechanisms of P. frutescens seeds, a total of 56 PfDof gene family members were identified from the genome and transcriptome data of P. frutescens and classified into four subfamilies according to sequence characteristics. All PfDofs contained highly conserved C₂-C₂ zinc finger domains, with gene duplication being the primary mechanism driving their evolution and expansion. Genes within the same subgroup exhibited similar gene structures and conserved motifs. The 56 PfDofs were predicted as unstable hydrophilic proteins, with α-helixes and random coils as their predominant structural components. The RNA-seq results revealed that 11 PfDofs exhibited differential expression during different developmental stages of P. frutescens seeds. RT-qPCR was performed to further validate the expression patterns of these 11 members across various tissue samples (root, stem, leaf, and flower) of P. frutescens and at different developmental stages of its seeds. The results showed that PfDof29 exhibited the highest expression level in seeds, which was consistent with the transcriptome data. Subcellular localization studies demonstrated that PfDof29 was localized to the nucleus and had a transcriptional activation activity. Overexpression of PfDof29 in Nicotiana tabacum resulted in a significant increase in total oil content of tobacco leaves, accompanied by reductions in starch and soluble sugar content, while the protein content remained unchanged. Additionally, the metabolic balance between saturated and unsaturated fatty acids in the transgenic tobacco leaves was altered, with a significant increase in α-linolenic acid content. The expression levels of the fatty acid desaturase genes NtFAD2, NtFAD3, and NtFAD8 were significantly upregulated. A yeast one-hybrid assay revealed that PfDof29 could directly bind to the promoter region of PfFAD8, thereby regulating its expression. This study provides an initial understanding of the regulatory mechanisms of PfDof transcription factors in the synthesis and accumulation of oil in P. frutescens. These findings offer new insights into the enhancement of oil content and quality of P. frutescens seeds.
Transcription Factors/physiology* ; Perilla frutescens/metabolism* ; Plant Proteins/metabolism* ; Gene Expression Regulation, Plant ; alpha-Linolenic Acid/biosynthesis* ; Lipids/biosynthesis* ; Seeds/genetics*

Objective:To explore the metabolic basis and related molecular mechanism of oral squamous cell carcinoma(OSCC)path-ogenesis.Methods:20 Chinese hamsters were divided into 2 groups(n=10).OSCC models were induced by dimethylbenzanthracene(DMBA)in 10 of the animals and the other 10 were used as the controls.LC-MS chromatography-mass spectrometry was used to iden-tify the metabolites in the buccal pouch,and multidimensional statistical analysis of the metabolites was performed with the orthogonal partial least squares discriminant analysis model.Variable Importance for the Projection(VIP)＞1 and P＜0.05 were used as the criteria to screen the differential metabolites between the 2 groups.KEGG pathway annotation and enrichment analysis for the metabolites were performed to screen the significantly differential pathways.Results:The hamster cheek pouches painted with 0.5％DMBA for 18 weeks were diffused with leukoplakia and loaded obvious papillary protrusions,which were diagnosed as OSCC by pathological examination.Lipids and lipid-like molecules were the main differential metabolites.Reprogramming of unsaturated fatty acid biosynthesis,cholesterol accumulation,enhanced catabolism of tryptophan,up-regulation of aspartate,increased synthesis of pyrimidine and purine,etc.were important metabolic features in the occurrence and development of OSCC.Conclusion:Molecular intervention targeting the related met-abolic pathways is expected to inhibit OSCC pathogenesis and progression.

Objective To assess the efficacy of transvaginal three-dimensional power Doppler ultrasound(3D-PDU)in predicting pregnancy outcomes after mild and moderate transcervical resection of adhesion(TCRA).Methods Sixty-six patients with mild and moderate intrauterine adhesions(IUA)who had fertility issues and underwent TCRA surgery in Tongde Hospital of Zhejiang Province from January 2020 to December 2021 were chosen,31 successful pregnancy patients were included in pregnancy group,and 35 unsuccessful pregnancy patients were included in non pregnancy group.3D-PDU examination was performed before and 3 months after surgery.The measurements of endometrial thickness(ED),uterine volume(EV),endometrial vasculogenesis index(VI),blood flow index(FI),and vasculogenesis-blood flow index(VFI)were taken.The pregnancy was followed up for 1 year to observe the prognostic value of the above related indicators on postoperative pregnancy.Results The ED,EV,VI,FI and VFI levels of patients post-operation were significantly higher than those before the operation(P＜0.05),the improvement of pregnant group patients was better than that of non pregnant group patients(P＜0.05),and area under the receiver operating characteristic curve were 0.80,0.84,0.68,0.65 and 0.88 respectively.Conclusion 3D-PDU can effectively predict the postoperative pregnancy outcome of TCRA in patients with mild to moderate IUA by measuring ED,EV,VI,FI,and VFI before and after surgery,providing reference for efficacy evaluation and guiding subsequent treatment.

BACKGROUND:In the treatment of skin trauma with active repair,tissue engineering techniques are needed to generate new tissue to replace necrotic tissue.Skin scaffolds have a good application prospect in the field of wound repair.Skin scaffolds need to present three-dimensional porous structures with certain mechanical strength to meet the needs of cell proliferation and division.However,the mechanical strength of the currently used gelatin-based biomaterials is weak and cannot meet the requirements of the use of skin scaffolds. OBJECTIVE:To study the 3D printing process used in the preparation of tissue engineering skin scaffolds by gelatin/oxidized nanocellulose composites,and focus on the relationship between the porosity and mechanical strength of the scaffolds prepared under different process parameters. METHODS:Oxidized nanocellulose whiskers at 10%concentration were extracted from Humulus scandens and then compounded with 5%gelatin to obtain gelatin/oxidized nanocellulose composites.The elastic modulus of gelatin and gelatin/oxidized nanocellulose composite was determined.Skin scaffolds were prepared by 3D printing extrusion molding using gelatin/oxidized nanocellulose composite as the base material.Mechanical and rheological properties of the composite were tested to determine extrusion molding parameters(filling gap 1.5-2.5 mm,uniform distribution of 0.1 mm;air pressure of 160-200 kPa),and the skin scaffold with a three-dimensional porous structure was prepared.The compressive performance of the skin scaffold was tested and compared with the finite element analysis results.The relationship between the filling gap and the porosity and mechanical strength of the scaffold was demonstrated. RESULTS AND CONCLUSION:(1)The elastic modulus of 5%gelatin was increased by 8.84 times by adding 10%oxidized nanocellulose whisker.A gel filament with a diameter of 1 mm was obtained by extrusion at the air pressure of 160 kPa.When the filling gap increased from 1.5 mm to 2.5 mm,the theoretical porosity of the scaffold increased from 33%to 60%,but the compressive strength decreased from 230 000 Pa to 95 000 Pa.(2)These findings showed that the skin scaffold with theoretical porosity of 50%and elastic modulus of 160 000 Pa was prepared by using 2 mm filling gap.The scaffold had a clear three-dimensional porous structure.

Objective:To explore the clinical efficacy and safety analysis of a novel laser localization technology assisted percutaneous puncture of trigeminal nerve microsphere capsule compression surgery for the treatment of primary trigeminal neuralgia.Methods:A retrospective selection was conducted on 63 patients with primary trigeminal neuralgia who underwent percutaneous puncture of the trigeminal nerve microsphere capsule compression surgery at the First Hospital of Hunan University of Chinese Medicine from January 2020 to December 2021. According to different surgical methods, they were divided into a new laser localization assisted puncture group (observation group) of 32 cases and a traditional barehanded localization puncture group (control group) of 31 cases. An analysis was conducted on the surgical time, puncture time, puncture frequency, intraoperative exposure to radiation, number of cases of poor balloon formation, and clinical efficacy within 6 months after surgery for two groups of patients. The prognosis of the patients was followed up at 6 months after surgery.Results:The surgical time, puncture time, puncture frequency, and intraoperative exposure of the observation group were all less than those of the control group, and the differences were statistically significant (all P<0.05). There was no statistically significant difference ( P>0.05) in the number of cases of poor balloon angioplasty between the observation group and the control group, as well as the pain score grading of the Barlow Neurological Institute (BNI) on the first day after surgery. Within 6 months after surgery, there was no statistically significant difference in the incidence of facial numbness, diplopia, masseter weakness, perilabial herpes, and recurrent pain between the two groups of patients (all P>0.05). Conclusions:Laser positioning technology can assist in precise puncture of the foramen ovale and accurate placement of balloons based on surgical experience, which helps to improve surgical safety, reduce postoperative complications and intraoperative radiation dose, and achieve satisfactory short-term follow-up results.

Objective:To explore the clinical characteristics and genetic variant of a patient with desminopathy manifesting with atypical symptoms.Methods:A patient who was admitted to the Department of Neurology of Jing′an District Central Hospital on February 24, 2021 was selected as the study subject. Clinical data, laboratory tests, muscle pathology, muscle magnetic resonance imaging (MRI) and genetic testing of the patient were retrospectively analyzed.Results:The patient had developed myalgia after lower limb activity, and gradually developed asymmetrical muscle weakness and atrophy of the lower limbs. Cardiac examination revealed atrioventricular block and decreased left ventricular diastolic function. Muscle MRI showed that semitendinosus, sartorius, gracilis, fibula, gastronemius and supinator muscles were selectively involved at the early stage. Muscle biopsy confirmed pathological changes of desmin positive myofibrils. Genetic testing revealed that the patient has harbored a c. 1024A>G (p.n342d) missense variant in exon 6 of the DES gene. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was rated as likely pathogenic (PS4_moderate+ PM2_supporting+ PP3_moderate+ PP1). Conclusion:Desmin disease has a great clinical heterogeneity. Postexercise myalgia of lower limbs is a rare clinical phenotype. For patients harboring the c. 1024A>G (p.n342d) variant of the DES gene, in addition to semitendinosus and fibula, Cardiac involvement is relatively insidious and easy to be ignored in clinic. Timely muscle MRI, muscle biopsy and gene detection will help the early diagnosis of the disease.

Objective To analyze the effects of intensive treatment with insulin pump and insulin aspart+insulin glargine 3+1.Methods Fifty patients with type 2 diabetes mellitus(T2DM)in our hospital were randomly divided into insulin aspart+insulin glargine(Asp+Gla)group and insulin pump(IP)group,with 25 cases in each group.Compare the general data,biochemical indicators,serum malondialdehyde(MDA),catalase(CAT),superoxide dismutase(SOD)and adverse reactions between the two groups,and calculate the insulin resistance index(HOMA-IR)and islet β function index(HOMA-β).Results HOMA-β,HDL-C,SOD and CAT in the two groups were higher than those before treatment(P<0.05),while FPG,2 hPG,GA,HOMA-IR,TG,TC,LDL-C and MDA were lower than those before treatment(P<0.05).Compared with Asp+Gla group after treatment,HDL-C,SOD and CAT in IP group increased(P<0.05),while FPG,2 hPG,GA,HOMA-β,HOMA-IR,TG,TC,LDL-C and MDA decreased(P<0.05).Conclusions Intensive treatment with insulin pump can more effectively regulate the level of glucose and lipid metabolism,improve insulin β function and reduce oxidative stress,which is safe and effective.

Objective:To investigate the application value of Pelvic Inflammation No.1 formula of transdermal meridian point-targeted drug delivery therapy in the treatment of patients with chronic pelvic inflammatory disease(damp-heat stasis type).Methods:All patients with chronic pelvic inflammatory disease(damp-heat stasis type)visited Danzhou Hospital of Traditional Chinese Medi-cine from August 2017 to March 2021 and were randomly grouped,57 patients in the control group were treated with simple Western medicine,and 57 patients in the observation group were combined with Pelvic Inflammation No.1 formula transdermal meridian target-ed transdermal medication,and the therapeutic efficacies were compared after 3 months of treatment.Results:After treatment,serum matrix metalloproteinase-9(MMP-9),D dimer(D-D),IL-6 and other indexes in the observation group were lower than those in the control group,and IL-10 was higher than that in the control group,the difference was statistically significant(P<0.05).After treat-ment,the carbohydrate antigen CA125 in the observation group was lower than that in the control group,and the CD4+/CD8+and CD4+cells in the observation group were higher than those in the control group(P<0.05).After treatment,the score of summary of health status surveys(SF-36)in the observation group was higher than that in the control group,and the total score of TCM symptoms was lower than that in the control group(P<0.05).The total effective rate in the observation group was 96.49%(55/57),which was higher than 82.46%(46/57)in the control group(P<0.05).Conclusion:For patients with chronic pelvic inflammatory disease(damp-heat stagnation type),combined with Pelvic Inflammation No.1 formula transdermal via acupoints targeted drug therapy can help to regu-late inflammatory factors,regulate immune function,and improve clinical efficacy.

Objective·To compare the effects and complications of carbon dioxide laser laryngectomy and open partial laryngectomy in the treatment of laryngeal carcinoma.Methods·Clinical data of 101 patients with stage Ⅰ-Ⅲ laryngeal squamous cell carcinoma admitted to Renji Hospital,Shanghai Jiao Tong University School of Medicine from January 2013 to January 2018 were selected and divided into open surgery group and minimally invasive group according to different surgical methods.The open surgery group received open partial laryngectomy(57 cases),and the minimally invasive group received carbon dioxide laser laryngectomy(44 cases).Follow-up visits were made by telephone and outpatient visits,and the operation time,postoperative laryngeal mucosa edema time,length of stay,5-year survival rate,complication rate and recurrence rate of the two groups were compared in the different clinical stages(stage Ⅰ,stage Ⅱ,and stage Ⅲ)and tumor locations(glottic type and non-glottic type).Results·The operation time,postoperative laryngeal mucosa edema time and length of stay of the minimally invasive group were shorter than those of the open surgery group(P＜0.05).Comparison of the 5-year survival rates of patients with the same clinical stage and glottic type between the two groups showed no statistically significant difference(both P＞0.05);while comparison of the 5-year survival rates of patients with non-glottic type between the two groups(62.5％in the open surgery group vs 0 in the minimally invasive group)showed a statistically significant difference(P＜0.05).In the minimally invasive group,there were 3 cases of complications,including 1 case of fever and 2 cases of lung infection;in the open surgery group,there were 14 cases of complications,including 5 cases of fever,7 cases of lung infection and 2 cases of pharyngeal fistula.The complication rate of patients in the minimally invasive group(6.82％,3/44)was lower than that in the open surgery group(24.60％,14/57),and the difference was statistically significant(P＜0.05);the complication rate of patients with stage Ⅱ and glottic type in the minimally invasive group was significantly lower than that in the open surgery group(P＜0.05).Comparing the recurrence rate of patients in the two groups(10.53％in the open surgery group vs 2.27％in the minimally invasive group),the difference was not statistically significant(P＞0.05).Conclusion·Compared with open partial laryngectomy,carbon dioxide laser laryngectomy has faster postoperative recovery and lower complication rate in the treatment of early and middle laryngeal malignant tumors,so it is worth promoting.

Chronic refractory wounds have been a challenge for clinical treatment because of their diverse causative factors and complex pathological processes, long healing times, and high treatment costs. The microenvironment of the wound surface includes the external microenvironment of the periwound surface, the internal microenvironment of the wound surface, and subsurface physiological structures. Research on clinical treatment strategies based on the microenvironment of chronic refractory wounds continues to innovate and make progress. Hydrogels have the advantages of high-water content, adjustable performance, good biocompatibility, and similarity to extracellular matrix. The ability of hydrogels to load drugs and their modification to confer excellent tissue adhesion, antibacterial, antioxidant, and modulation of inflammatory factor expression can be used to achieve a multi-factor response and modulation of the physical, chemical, and biological aspects of the trauma microenvironment. Therefore, hydrogels have outstanding advantages and clinical application prospects in the repair of chronic, difficult-to-heal wounds. In this review paper, the characteristics and etiology of chronic refractory wounds were introduced, and the classification of microenvironment-responsive hydrogels for chronic refractory wounds and their application in the repair of refractory wounds were reviewed. Besides, the shortcomings of current hydrogels were discussed, and an outlook was proposed.