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1.
Chinese Journal of Biotechnology ; (12): 4887-4900, 2023.
Article in Chinese | WPRIM | ID: wpr-1008066

ABSTRACT

This study aimed to explore the effect of miR-23b-3p on the differentiation of goat intramuscular preadipocytes, and to confirm whether miR-23b-3p plays its roles via targeting the PDE4B gene. Based on the pre-transcriptome sequencing data obtained previously, the miR-23b-3p, which was differentially expressed in goat intramuscular adipocytes before and after differentiation, was used as an entry point. real-time quantitative-polymerase chain reaction (qPCR) was used to detect the expression pattern of miR-23b-3p during the differentiation of goat intramuscular preadipocytes. The effects of miR-23b-3p on adipose differentiation and adipose differentiation marker genes were determined at the morphological and molecular levels. The downstream target genes of miR-23b-3p were determined using bioinformatics prediction as well as dual luciferase reporter assay to clarify the targeting relationship between miR-23b-3p and the predicted target genes. The results indicated that overexpression of miR-23b-3p reduced lipid droplet accumulation in goat intramuscular adipocytes, significantly down-regulated the expression levels of adipogenic marker genes AP2, C/EBPα, FASN, and LPL (P < 0.01). In addition, the expressions of C/EBPβ, DGAT2, GLUT4 and PPARγ were significantly downregulated (P < 0.05). After interfering with the expression of miR-23b-3p, lipid droplet accumulation was increased in goat intramuscular adipocytes. The expression levels of ACC, ATGL, AP2, DGAT2, GLUT4, FASN and SREBP1 were extremely significantly up-regulated (P < 0.01), and the expression levels of C/EBPβ, LPL and PPARγ were significantly up-regulated (P < 0.05). It was predicted that PDE4B might be a target gene of miR-23b-3p. The mRNA expression level of PDE4B was significantly decreased after overexpression of miR-23b-3p (P < 0.01), and the interference with miR-23b-3p significantly increased the mRNA level of PDE4B (P < 0.05). The dual luciferase reporter assay indicated that miR-23b-3p had a targeting relationship with PDE4B gene. MiR-23b-3p regulates the differentiation of goat intramuscular preadipocytes by targeting the PDE4B gene.


Subject(s)
Animals , MicroRNAs/metabolism , Goats/genetics , PPAR gamma/metabolism , Adipogenesis/genetics , Cell Differentiation/genetics , Luciferases , RNA, Messenger
2.
Chinese Journal of Biotechnology ; (12): 2695-2705, 2023.
Article in Chinese | WPRIM | ID: wpr-981226

ABSTRACT

The aim of this study was to clone the goat RPL29 gene and analyze its effect on lipogenesis in intramuscular adipocytes. Using Jianzhou big-eared goats as the object, the goat RPL29 gene was cloned by reverse transcription-polymerase chain reaction (RT-PCR), the gene structure and expressed protein sequence were analyzed by bioinformatics, and the mRNA expression levels of RPL29 in various tissues and different differentiation stages of intramuscular adipocytes of goats were detected by quantitative real-time PCR (qRT-PCR). The RPL29 overexpression vector pEGFP-N1-RPL29 constructed by gene recombination was used to transfect into goat intramuscular preadipocytes and induce differentiation. Subsequently, the effect of overexpression of RPL29 on fat droplet accumulation was revealed morphologically by oil red O and Bodipy staining, and changes in the expression levels of genes related to lipid metabolism were detected by qRT-PCR. The results showed that the length of the goat RPL29 was 507 bp, including a coding sequence (CDS) region of 471 bp which encodes 156 amino acid residues. It is a positively charged and stable hydrophilic protein mainly distributed in the nucleus of cells. Tissue expression profiling showed that the expression level of this gene was much higher in subcutaneous adipose tissue and inter-abdominal adipose tissue of goats than in other tissues (P < 0.05). The temporal expression profile showed that the gene was expressed at the highest level at 84 h of differentiation in goat intramuscular adipocytes, which was highly significantly higher than that in the undifferentiated period (P < 0.01). Overexpression of RPL29 promoted lipid accumulation in intramuscular adipocytes, and the optical density values of oil red O staining were significantly increased (P < 0.05). In addition, overexpression of RPL29 was followed by a highly significant increase in ATGL and ACC gene expression (P < 0.01) and a significant increase in FASN gene expression (P < 0.05). In conclusion, the goat RPL29 may promote intra-muscular adipocyte deposition in goats by up-regulating FASN, ACC and ATGL.


Subject(s)
Animals , Lipogenesis/genetics , Adipogenesis/genetics , Goats/genetics , Adipocytes , Cell Differentiation/genetics , Sequence Analysis , Cloning, Molecular
3.
Chinese Journal of Biotechnology ; (12): 1696-1709, 2023.
Article in Chinese | WPRIM | ID: wpr-981164

ABSTRACT

The purpose of this study was to clone and characterize the ZFP36L1 (zinc finger protein 36-like 1) gene, clarify its expression characteristics, and elucidate its expression patterns in different tissues of goats. Samples of 15 tissues from Jianzhou big-eared goats, including heart, liver, spleen, lung and kidney were collected. Goat ZFP36L1 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR), then the gene and protein sequence were analyzed by online tools. Quantitative real-time polymerase chain reaction (qPCR) was used to detect the expression level of ZFP36L1 in intramuscular preadipocytes in different tissues and adipocytes of goat at different differentiation stages. The results showed that the length of ZFR36L1 gene was 1 224 bp, and the coding sequence (CDS) region was 1 017 bp, encoding 338 amino acids, which was a non-secretory unstable protein mainly located in nucleus and cytoplasm. Tissue expression profile showed that ZFP36L1 gene was expressed in all selected tissues. In visceral tissues, the small intestine showed the highest expression level (P < 0.01). In muscle tissue, the highest expression level was presented in longissimus dorsi muscle (P < 0.01), whereas the expression level in subcutaneous adipose tissue was significantly higher than that in other tissues (P < 0.01). The results of induced differentiation showed that the expression of this gene was up-regulated during adipogenic differentiation of intramuscular precursor adipocytes (P < 0.01). These data may help to clarify the biological function of the ZFP36L1 gene in goat.


Subject(s)
Animals , Goats/genetics , Amino Acid Sequence , Liver , Cloning, Molecular
4.
Chinese Journal of Gastrointestinal Surgery ; (12): 1165-1169, 2019.
Article in Chinese | WPRIM | ID: wpr-800468

ABSTRACT

Objective@#To compare the efficacy and safety of two procedures in the treatment of hemorrhoid: the procedure for prolapse and hemorrhoids (PPH) and stapled transanal rectal resection (STARR).@*Methods@#A retrospective cohort research was conducted. Clinical data of 263 patients undergoing the first elective surgery for grade IV hemorrhoids by the same team of surgeons at our department from January 2015 to December 2016 were analyzed retrospectively, while those had other anorectal diseases, emergency surgery, inflammatory bowel disease, tumor and incomplete clinical data were excluded. PPH was performed in 129 patients and STARR was performed in 134 patients. PPH procedure: a circular purse 2-0 string suture was made at 4 cm above the dentate line; in accordance with the standard protocol, the PPH circular stapling devicewas introduced; the suture was closed, and a pull-through followed; the traction was continued; the stapler was fired; the prolapsed mucosa and submucosa were removed. STARR procedure: 3-5 needles were sutured in the anterior rectal mucosa, protecting the posterior wall mucosa; with the help of a finger the PPH stapler was inserted into the vaginal lumen; the sutures were hooked from both sides of the stapler to maintain traction; according to the disease condition, the suturewas tightened appropriately; stapler was screwed and activated; the anterior wall mucosa was removed; the joint of the both ends of anastomosis was cut; the posterior wall mucosa was removed as well. The short-term efficacy, surgical safety and prognosis of the two groups were compared.@*Results@#There were 67 males (51.9%) in the PPH group and 57 (42.5%) males in the STARR group. The median age of the two groups was 51.0 (22.0, 80.0) years and 49.0 (24.0, 74.0) years, respectively. There were no significant differences in the baseline data between the two groups (all P>0.05). No significant differences in the intraoperative bleeding, length of hospital stay, postoperative analgesic drug use, postoperative bleeding, postoperative infection, etc. were found between two groups (all P>0.05). As compared to PPH group, STAAR group had longer operation time and higher hospitalization cost with significant differences [(44.0±19.3) minutes vs. (26.3±8.5) minutes, t=9.701, P=0.001; (11 047±473) yuan vs. (7674±309) yuan, t=32.826, P=0.001]. One case in STAAR group developed rectovaginal fistula. The median follow-up period of the whole group was 40 (33, 52) months. A total of 108 cases in STARR group and 114 cases in PPH group completed the follow-up. The 3-year disease-relapse rate was 0 in STARR group and 4.2% in PPH group (P=0.042).@*Conclusion@#STARR procedure can improve the prognosis in the treatment of grade IV hemorrhoid, but attention should be paid to the development of complications.

5.
International Journal of Traditional Chinese Medicine ; (6): 938-942, 2018.
Article in Chinese | WPRIM | ID: wpr-693699

ABSTRACT

Objective To systematically evaluate the effectiveness and safety of acupuncture intervention in gastrointestinal function after gynecilogical and obstetrical operations. Methods Databases of SinoMed (1997-2017), VIP (1997-2017), Wanfang (1997-2017), CNKI (2007-2017), PubMed (1997-2017) and Cochrane Library (2017 fourth) were searched by computers (supplemented by manual searching). The randomized controlled trials were included on the gastrointestinal function recovery after acupuncture intervention in gynecilogical and obstetrical operations. The data were extracted independently and cross-checked by two evaluators. Then the quality was evaluated according to the Cochrane system assessor manual 4.2.8, and RevMan 5.3 software was used for meta-analysis. Results A total of 1192 patients were included in 14 randomized controlled trials. Meta-analysis showed that the effective rate [OR=6.09, 95% CI (2.91, 12.75), P<0.01], the time to first passage of feces [MD=-14.72,95% CI(-16.00, -13.44),P<0.01], the time to first bowel motion [MD=-8.81, 95% CI (-10.34, -7.28), P<0.01] and the time to first flatus [MD=-11.84, 95% CI (-15.31, -8.36), P<0.01] in the acupuncture group were higher than those in control group. Conclusions Acupuncture intervention in gynecilogical and obstetrical operations was safe and effective for gastrointestinal function recovery of patients. However, the above conclusions are required to have further validation with more highly qualified randomized controlled trials for the limited number of literature and the low quality of some studies.

6.
Chinese Critical Care Medicine ; (12): 484-488, 2015.
Article in Chinese | WPRIM | ID: wpr-463680

ABSTRACT

Objective To explore the effect of selective gut decontamination in regulation of inflammatory reaction compared with rhubarb and glycerine enema for catharsis in patients with systemic inflammatory response syndrome ( SIRS ), and to discuss its mechanisms. Methods A prospective randomized controlled trial was conducted. Fifty-seven patients with SIRS admitted to Department of General Surgery of Aviation General Hospital from June 2009 to June 2014 were enrolled. The patients were randomly divided into rhubarb decontaminate group, traditional decontaminate group and blank control group, with 19 cases in each group. Besides the treatment for primary disease, including anti-infection, operation, alleviate pain, nutritional support, and maintaining water and electrolyte balance, the patients in rhubarb decontaminate group received aqueous extract from rhubarb 15-20 g by gastric tube, enema, or peros, twice a day;and those in traditional decontaminate group received glycerine enema or glycerol enema, twice a day; while no gavage or enema was prescribed in blank control group. Peripheral blood was collected before and 72 hours after treatment. Enzyme linked immunosorbent assay ( ELISA ) was used to determine the concentration of lipopolysaccharide ( LPS ) and inflammatory mediators. Results Compared with blank control group and traditional decontaminate group, the levels of interleukins ( IL-1, IL-8 ), LPS, platelet activating factor ( PAF ), tumor necrosis factor-α( TNF-α), andγ-interferon ( IFN-γ) before treatment was similar to that of rhubarb decontaminate group [ IL-1 ( ng/L ): 53.154±5.783, 50.564±5.771, 51.082±6.403, F = 0.994, P = 0.377; IL-8 ( ng/L ): 70.492±6.146, 68.376±6.112, 68.673±8.384, F=0.514, P=0.601;LPS (μg/L ):11.630±2.449, 10.858±2.307, 10.463±2.145, F = 1.261, P = 0.291; PAF (μg/L ): 4.173±0.395, 4.051±0.362, 4.078±0.487, F = 0.446, P = 0.642; TNF-α( ng/L ):132.498±10.772, 129.735±12.881, 127.207±11.514, F=0.963, P=0.388;IFN-γ(μg/L ):45.645±4.558, 43.692±5.578, 43.767±5.028, F = 0.904, P = 0.411 ]. The above parameters after treatment were significantly lower than those before treatment in three groups. The effect on the LPS and pro-inflammatory factors of the rhubarb decontaminate group was more obvious than that of the blank control group and traditional decontaminate group [ LPS (μg/L ): 7.571±1.113 vs. 9.008±1.904, 8.874±1.808, F = 4.416, P = 0.017; IL-1 ( ng/L ): 45.309±3.563 vs. 48.731±4.466, 46.112±4.322, F = 3.557, P = 0.035; IL-8 ( ng/L ): 60.492±5.346 vs. 65.553±5.384, 63.437±5.462, F = 4.213, P = 0.020; PAF (μg/L ): 3.519±0.250 vs. 3.832±0.356, 3.766±0.309, F = 5.450, P = 0.007; TNF-α ( ng/L ): 114.988±8.772 vs. 123.230±10.433, 118.534±9.519, F = 3.525, P = 0.036; IFN-γ(μg/L ):38.683±3.190 vs. 41.831±4.122, 39.161±3.972, F=3.820, P=0.028 ]. Conclusion The usage of selective gut decontamination can inhibit the release of endotoxin and inflammatory mediators in patients with SIRS, and it will get a better effect using rhubarb, and the mechanism may be related to the protection of intestinal mucosal barrier function.

7.
Chinese Journal of General Surgery ; (12): 272-275, 2015.
Article in Chinese | WPRIM | ID: wpr-468693

ABSTRACT

Objective To probe the association between possible single nucleotide polymorphism (SNP) in the FGB promoter region and idiopathic deep venous thrombosis.Methods A prospective analysis was performed in both IDVT group and control group (120 cases each) followed by a duplex examination using gene sequencing technique and restriction fragment length polymorphism (RFLP) in the promoter region of fibrinogen gene β.Possible SNPs in this region were detected arranged before HardyWeinberg equilibrium test and Linkage disequilibrium (LD) analyses.Ultimately,we compared the genotype frequencies between the two groups and undertook a multiple Logistic regression.Results Six kinds of SNPs were determined in the promoter region of β-fibrinogen gene:-148C/T,-249C/T,-455G/A,-854G/A,-993C/T and-1420G/A.A stronger linkage disequilibrium was confirmed between-993C/T and -455G/A (r2 =0.699) ;-993C/T and-148C/T (r2 =0.509) ;-455G/A and-148C/T (r2 =0.556).Statistical differences of genotype frequencies between two groups were observed in-148C/T,-249C/T,-455G/A and-1420G/A polymorphisms (all P < 0.05).Conclusions The risk of IDVT was 4.579 times higher with every 1 g/L increase of fibrinogen concentration.Allele-148T,-455G and-1420A are IDVT risk factors.-993C/T may indirectly affect IDVT through linkage disequilibrium with-455G/A and-148C/T.

8.
Chinese Journal of Gastrointestinal Surgery ; (12): 1106-1110, 2014.
Article in Chinese | WPRIM | ID: wpr-235006

ABSTRACT

<p><b>OBJECTIVE</b>To compare the safety and efficacy between extralevator abdominoperineal excision(ELAPE) and traditional abdominoperineal excision(APE) in patients with low rectal cancer.</p><p><b>METHODS</b>From January 2011 to December 2013, 61 patients undergoing abdominoperineal excision for low rectal cancer at the Department of Gastrointestinal Surgery, Peking University People's Hospital were enrolled. The safety and efficacy of two procedure groups, ELAPE group (33 patients) and traditional APE group (28 patients) were reviewed retrospectively.</p><p><b>RESULTS</b>Less intraoperative blood loss [(201 ± 147) ml vs. (343 ± 308) ml, P=0.022], shorter exhaust time [(3.8 ± 1.5) d vs. (4.6 ± 1.6) d, P=0.039] and lower perineal wound complication rate (9.1% vs. 25.0%, P=0.042) were observed in the ELAPE group as compared to the traditional APE group. However, longer operation time [(292 ± 46) min vs. (256 ± 28) min, P=0.008] and perineal drainage removal time [(11.1 ± 4.8) d vs. (7.1 ± 2.7) d, P<0.01] were noted in the ELAPE group than those in the traditional APE group. Number of lymph node retrieved and positive lymph node retrieved was not significantly different between two groups. The ELAPE group had lower rate of IOP (intraoperative perforation) (6.1% vs. 17.9%, P=0.055), but no significant difference was found. There were no significant differences in bowel movement, diet restoring time, average perineal drainage, postoperative hospitalization time and removing perineal stitches time between the two groups (all P>0.05).</p><p><b>CONCLUSION</b>ELAPE possesses less intraoperative blood loss and lower perineal wound complication rate than traditional APE. ELAPE is associated with better safety and efficacy than traditional APE.</p>


Subject(s)
Humans , Digestive System Surgical Procedures , Operative Time , Rectal Neoplasms , General Surgery , Retrospective Studies
9.
Chinese Journal of General Surgery ; (12): 93-95, 2013.
Article in Chinese | WPRIM | ID: wpr-432331

ABSTRACT

Objective To explore the clinical outcomes of laparoscopic appendectomy in acute and chronic appendicitis,and sum up the surgical experiences of lapaproscopic appendectomy.Method In this study 129 cases of appendicitis at the Department of Gastroentrological Surgery,Peking University People's Hosptial were collected retrospectively from June 2008 to December 2009.The clinical results of laparoscopic and open procedures for acute appendicitis and the outcomes of laparoscopic operation for acute and chronic appendicitis were compared.Results For acute appendicitis,the length of hospitalization [(4.8 ± 2.6) d vs.(7.0 ± 1.3) d,t =0.679,P =0.006] was significantly shorter in laparoscopic group than that in open surgery group.In addition,the mean length of operation time [(77 ± 33) min vs.(55 ± 23) min,t =3.431,P <0.01] were longer,postoperative first passing flatus [(2.3 ± 1.2) d vs.(1.4 ±0.9) d,t =4.665,P <0.01] and oral intake [(2.3 ± 1.4) d vs.(1.2 ±0.6) d,t =4.517,P<0.01] were later for acute appendicitis patients than for chronic appendicitis in laparoscopic group.Conclusions Laparoscopic appendectomy for acute appendicitis is a safe and effective procedure,though it might cause more postoperative complications such as intra-abdominal abscess and small intestinal obstruction in patients with acute appendicitis.

10.
Chinese Journal of General Surgery ; (12)1993.
Article in Chinese | WPRIM | ID: wpr-526954

ABSTRACT

Objective The purpose of this study is to investigate the factors associated with incisional surgical site infection (SSI) in colorectal cancer. Methods We retrospectively analyzed impacting factors of SSI including age, gender, BMI, diabetes mellitus, operation, albumin levels, hemoglobin levels, tumor staging, and length of stay post-operation in 327 colorectal patients from 2001 to 2005. Results The incidences of SSI and fat liquefaction were 2. 4%(8/327) and 12. 2%(40/327) respectively. The incidences of SSI in patients with obesity and diabetes mellitus increased significantly (16. 8% vs. 10. 0% ,28. 6% vs. 13. 7%). The length of stay in patients with SSI prolonged significantly (29.0 d vs. 15.8 d). Conclusion Obesity and diabetes mellitus are the most important factors associated with SSI in colorectal cancer patients.

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