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 investigate the expressions and clinical significances of histone marks H3K9me3 and H3K27me3 in colorectal cancer.Methods:A retrospective case-control study was conducted. The clinical data of 98 patients with colorectal cancer in Shanxi Province Cancer Hospital from May 2008 to July 2017 were retrospectively analyzed, including 35 patients in the non-metastatic operation-only group, 29 patients in the synchronous hepatic oligometastasis group and 34 patients in the extensive metastasis group, and 33 patients with benign colorectal lesions who underwent colonoscopy in 2017 were selected as the control group. Immunohistochemical assay was used to detect the expressions of H3K9me3 and H3K27me3 proteins in each group, and the expressions of H3K9me3 and H3K27me3 proteins in colorectal cancer patients with different clinicopathological features were analyzed. Kaplan-Meier method was used for survival analysis and log-rank test was performed.Results:The positive expression rate of H3K9me3 protein in colorectal cancer group was 11.2% (11/98), which was lower than that in control group [60.6% (22/33)] ( χ2 = 33.33, P < 0.001); the positive expression rate of H3K27me3 protein in colorectal cancer group was 10.6% (13/98), which was lower than that in control group [97.0% (32/33)] ( χ2 = 76.70, P < 0.001). The positive expression rates of H3K9me3 protein were 60.6% (20/33), 17.1% (6/35), 10.3% (3/29) and 5.9 % (2/34) in the control group, the non-metastatic operation-only group, the synchronous hepatic oligometastasis group and the extensive metastasis group, respectively, and the difference was statistically significant ( χ2 = 26.10, P < 0.001); the positive expression rates of H3K27me3 protein were 97.0% (32/33), 14.3% (5/35), 20.7% (6/29) and 5.9% (2/34), respectively, and the difference was statistically significant ( χ2 = 44.16, P < 0.001). The positive expression rate of H3K27me3 in colorectal cancer tissues of patients with lymph node metastasis degree ≤0.2 was higher than that of patients with lymph node metastasis degree >0.2 [22.4% (11/49) vs. 4.2% (2/48), χ2 = 6.98, P = 0.008]. The median overall survival (OS) time of H3K9me3 positive and negative colorectal cancer patients was 77.0 months (95% CI: 10.6-143.3 months) and 34.0 months (95% CI: 25.5-42.5 months), respectively, and there was no significant difference in OS between the two groups ( P = 0.078). The median OS time of H3K27me3 positive and negative colorectal cancer patients was 39.0 months (95% CI: 15.3- 62.7 months) and 34.0 months (95% CI: 24.3-43.7 months), respectively, and there was no significant difference in OS between the two groups ( P = 0.524). Conclusions:The expressions of H3K9me3 and H3K27me3 in colorectal cancer tissues are lower than those in colorectal benign lesions, and gradually decrease with occurrence of liver metastasis and extensive metastasis. H3K9me3 and H3K27me3 may be potential cancer suppressor factors.

Objective To screen Oxalobacter formigenes (OxF) from fresh feces of healthy adults,and study its effect on the the prevention of calcium oxalate kidney stones.Methods OxF was screened and cultured from fresh feces of healthy adults.The rat model of calcium oxalate stone was established by esophageal gavage of 0.8％ of ethylene glycol.Rats were divided into a control group and four groups of rats with ethylene glycol-induced calcium oxalate kidney stones according to random number table.Three groups were treated with 106 CFU,107 CFU,108 CFU viable OxF every day,respectively,for 4 weeks.The blood and 24-hour urine samples were collected to detect the serum creatinine,urea nitrogen,serum and urine calcium,phosphorus,magnesium and urine oxalate every week.At the end of the 4th week,the rats were sacrificed and the kidney tissues were stained with HE and Yasue.The deposition and content of calcium oxalate crystals were observed under a light microscope.Results The bacteria strain isolated from fresh feces of healthy adults was 100％ as same as the known ATCC35274 bacteria strain,which means the strain screened is OxF.Among the 5 groups,there were no significant differences in body weight,Scr,BUN,serum calcium,blood magnesium,blood phosphorus,urinary magnesium and urinary phosphorus.The 24-hour urinary calcium excretion in the model group was significantly lower than that of the control group (P ＜ 0.05).After intervention with OxF solution,the 24-hour urinary calcium excretion in the 108 CFU OxF group was significantly higher than that in the model group (P ＜ 0.05),while there was no significant difference between the other intervention groups and the model.The oxalic acid excretion of 106 CFU OxF group and 107 CFU OxF group was lower than that of the model,but the difference did not reach statistical significance (P＞ 0.05).The 24 h oxalic acid excretion in the 108 CFU OxF group was significantly lower than that of the model at the end of first week (P ＜ 0.05),and continued to decrease for the next 3 weeks.After 4 weeks of intervention,no crystal formation was observed in the control group under the deflection microscope,but a large amount of calcium oxalate crystals were formed in the renal cortex and renal medulla.The crystals were piled up and connected to each other.Yasue staining coincided with the calcium oxalate crystal in the same part of the kidneys.Compared with the model,there was no significant change in the score of calcium oxalate crystal in the kidneys of 106 CFU OxF group and 107 CFU OxF group,while the score of calcium oxalate crystal in the kidneys of 108 CFU OxF group was significantly lower (P ＜ 0.05).Conclusions OxF are successively screened from healthy adults.Daily administration of 108 CFU OxF can safely and effectively reduce the urinary oxalic acid excretion,prevent the formation of calcium oxalate crystals and inhibit the formation of stones in kidneys of rats.

Objective:To determine the effects of Jinlong capsule combined with interventional therapy on the immune functions of primary hepatocellular carcinoma patients. Methods:Sixty randomly selected cases of clinically diagnosed primary hepatocellular carcinoma were divided into the observation group and the control group. Three days after operation, the observation group was given four Jinlong capsules three times a day for 30 days (one treatment). Meanwhile, the control group received interventional therapy after the operation. One to four days following one treatment, peripheral blood specimens were collected from the two groups to determine the cellular immune function indices. Results:The cell numbers (mean) of the peripheral blood components CD3, CD4, NK, SIL-2R, TSGF, and SIL-2R and the CD4/CD8 ratio in the observation group showed no significant difference before and after treatment. In the control group, these indices were significantly different before and after treatment. Conclusion:The Jinlong capsule facilitates the cellu-lar immunity recovery of patients with primary hepatocellular carcinoma after interventional therapy.

Objective To explore the tumor markers for the diagnosis of primary liver carcinomas (PLC) by detecting the serum protein spectrum differently expressed between PLC patients and healthy controls. Methods We detected the serum protein spectrum in 50 PLC patients and 50 healthy controls using surface enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) technique and find the significant protein peaks. The serum Alpha-fetoprotein (AFP) levels in all 100 serum samples were also measured by ELISA. Results The protein peaks, which could discriminate healthy individuals from PLC patients, were detected. Four protein molecules (3354.71, 8825.80, 4345.08, 13 715.01) had a significant difference between PLC patients and the normal controls (P <10-5), indicating that these protein molecules might be a potential marker for PLC. The specificity and sensitivity of SELDI-TOF-MS were 94% and 90% respectively. Sixteen PLC patients were AFP positive and the sensitivity was 54%(27/50). Conclusion With a high specificity and sensitivity, the detection of serum protein spectrum can be performed easily and quickly by SELDI-TOF-MS technique, which provides a serological way in identifying PLC and most likely to benefit from AFP strategies.