This study reported the obtainment of the full-length cDNA of Salvia miltiorrhiza hairy roots (Abbr: SmHDR, GenBank number: JX233817), via extracting Salvia miltiorrhiza hairy roots total RNA, designing specific primers according to the transcriptome data and using the RACE strategy, and then analyzed it with bioinformatics approaches. On this basis, using the real-time PCR to detect SmHDR gene expression after Ag+ induction, and testing tanshinones contents of corresponding samples by UPLC. SmHDR has 1 647 nucleotides, and an open reading frame (ORF) encoding a protein of 463 amino acid residues. The deduced protein has isoelectric point (pI) of 5.72 and a calculated molecular weight about 51.88 kD. In the secondary structure, the percentage of alpha helix, beta turn and random coil were 35.64%, 20.30% and 44.06%, respectively. Sequence alignment and phylogenetic analysis demonstrated that SmHDR had relative close relationship to the HDR of Picrorhiza kurrooa, similar to HDR from other species of plants. Real time PCR results indicated that elicitor of Ag+ stimulated the increase of mRNA expression of SmHDR. At the same time, results of ultra performance liquid chromatography (UPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy roots of Salvia miltiorrhiza were increased dramatically at 12 h after treated with Ag+, and then decreased significantly. This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by Ag+. The content of tanshinones was gradually raised, and it had an obvious increase at 120 h. The bioinformatics analysis and gene expression indicated that SmHDR might be involved in tanshinones biosynthesis, which laid the foundation for further study of secondary metabolic regulation mechanism of tanshinones.

Objective:To explore the expression of long non-coding RNA LINC00630 in bladder cancer cell lines, and to explore the effect of interference with its expression in vitro on the proliferation and migration of bladder cancer cells.Methods:Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of LINC00630 in bladder cancer cell lines 5637, BIU-87, T24, J82 and normal bladder epithelial cell line SV-HUC-1. The bladder cancer cell line with the highest LINC00630 expression was selected for follow-up experiments, then the cell line infected with the control lentivirus was used as the control group, and the cell line infected with the lentivirus that could interfere with the expression of LINC00630 was used as the experimental group. qRT-PCR was used to detect the expression of LINC00630 in the two groups of cells. MTS method and cell scratch test were used to detect the proliferation and migration abilities of cells in the two groups. qRT-PCR was used to detect the expression of neuregulin 1 (NRG1) mRNA in the two groups of cells, and Western blot was used to detect the expressions of NRG1 protein, cell proliferation-related proteins (cyclin D3 and CDK2) and cell migration-related proteins (Vimentin and N-cadherin) in the two groups of cells.Results:Compared with SV-HUC-1 cells (1.05±0.17), the expression of LINC00630 was significantly increased in all bladder cancer cell lines (all P < 0.01), and the expression was highest in J82 cells (relative expression 5.83±0.42). Compared with J82 cells of the control group, the expression of LINC00630 in J82 cells of the experimental group decreased (0.18±0.02 vs. 1.00±0.05, t=14.36, P < 0.01); from day 2 of transfection, the cell proliferation activity of the experimental group was lower than that of the control group (all P < 0.05). The cell scratch closure rate of the experimental group was lower than that of the control group [(27.4±7.1)% vs. (66.0±5.4)%, t = 4.31, P < 0.01]. Therelative expression of NRG1 mRNA in the experimental group was lower than that in the control group (0.34±0.03 vs. 1.07±0.24, t = 2.99, P < 0.05). Compared with the control group, the expressions of NRG1 protein, cell proliferation-related proteins and cell migration-related proteins in the experimental group were reduced. Conclusions:LINC00630 is up-regulated in bladder cancer cell lines, and interference with LINC00630 may inhibit the proliferation and migration of J82 cells by down-regulating the expression of NRG1 gene. LINC00630 may be a new molecular target for the treatment of bladder cancer.

OBJECTIVETo compare the success rate, postoperative complications and cosmetic appearance of the perinatal flap (Mathieu) and the tubularized incised-plate (Snodgrass) urethroplasty in distal hypospadias repair.

METHODSBetween March 2001 and April 2004, Snodgrass-type repair was done in 36 patients and Mathieu-type repair in 43 patients with primary hypospadias either with or without minimal chordee.

RESULTSThe mean duration of surgery was significantly shorter for the Snodgrass procedure than for the Mathieu repair. With 3-10 months follow-up, the total numbers of complications of Snodgrass repair and Mathieu repair were 8 (22.2%) and 16 (37.2%) (P > 0.05), the fistula formation rates 8.3% and 25.6%, the meatal stricture formation rates 5.6% and 2.3%, and the wound dehiscence rates 8.3% and 2.3%, respectively. Flap necrosis was seen in 3 cases in the Mathieu group. Cosmetic appearance was highly satisfactory in the Snodgrass group.

CONCLUSIONThe fistula and flap necrosis rates were lower and the surgery more convenient with the Snodgrass urethroplasty, which also had a better cosmetic outcome. The Snodgrass technique is recommended as a primary treatment to substitute Mathieu repair for distal hypospadias with or without minimal chordee.

Child ; Child, Preschool ; Follow-Up Studies ; Humans ; Hypospadias ; surgery ; Male ; Penis ; Surgical Flaps ; Urethra ; blood supply ; Urologic Surgical Procedures, Male ; methods

Objective: To explore the application of penile base staggered flap in the operation of severe hypospadias with penoscrotal transposition. Methods: Twenty-three cases with severe hypospadias with penoscrotal transposition were corrected by urethroplasty and staggered flap plasty in our department from May 2013 to June 2016. After urethroplasty, the skin at the junction of the penile base and scrotum was transversely cut by 1.5-2.0 cm, and then sutured with Z-plasty to reconstruct the angle of the penis and scrotum. Results: After the operation, infection and cracking at the suture of the staggered flap occurred in one child and was cured after dressing change and symptomatic treatment. Two cases suffered from urine leakage at the suture of the staggered flap. One of the cases realized self-healing and the other received a repair operation for urinary fistula. Three children suffered coronary sulcus fistula and was cured by urethroplasty. All the patients were followed up from 10 to 30 months with normal penis scrotal position and satisfactory appearance. Conclusions The application of penile base staggered flap can be used for severe hypospadias in one stage with penoscrotal transposition surgery. It is a simple and satisfactory method for the treatment of hypospadias with penoscrotal transposition.

Objective To explore the diagnosis,treatment options and outcomes of patients with ureteropelvic junction obstruction(UPJO)and ureterovesical junction obstruction(UVJO). Methods Clinical records of 12 children with UPJO and UVJO who received treatment in Children′s Hospital of Nanjing Medical University from April 2008 to December 2015 were reviewed. Presentations included prenatal hydronephrosis and symptoms caused by urinary tract in-fections. Ultrasonography,intravenous pyelography,magnetic resonance imaging and renography were performed before surgery. Results Four patients underwent nephrostomy,followed by pyeloplasty and eventually,ureteroneocystostomy. Five patients underwent heterochronic pyeloplasty plus ureteroneocystostomy. Two patients received pyeloplasty only. And another patient underwent heterochronic nephrostomy plus ureteroneocystostomy. The 12 patients received a 2. 0 months to 2. 5 years′ follow - up. Recurrent urinary tract infection and abdominal pain were relieved after the treatment. Ultrasonography showed hydronephrosis reduced obviously after the surgery. Conclusions UVJO patients with unparallel hydronephrosis should be considered with the coexistence of UPJO. Retrograde pyelography(RPG)is recommended for the diagnosis. And the treatment includes both pyeloplasty and ureteroneocystostomy. The patients diagnosed with UPJO should be first managed with pyeloplasty or nephrostomy instead of RPG. Subsequently anterograde pyelography can be performed to decide if the patients need additional ureteroneocystostomy.

Objective: To summarize the diagnosis and prognosis of adrenocortical carcinoma in children. Methods: Medical records of 10 pediatric patients with adrenocortical cancer treated in Children's Hospital of Nanjing Medical University from September 2010 to March 2018 were retrospectively reviewed, and their clinical manifestation, laboratory, auxiliary, and pathological examination results, and prognosis were analyzed. Results: The average age of all patients (5 male and 5 female pediatric patients) was (53.3+48.4) months. Clinical mani-festations included abdominal mass, hypertension, cortisol level, and peripheral precocious puberty. Laboratory tests showed that lactate dehydrogenase, neuroenolase, and cortisol levels were high in some patients, and sex hormones were abnormal. Auxiliary examination using the abdominal computed tomography enhancement showed irregular an adrenal gland, uneven density, necrosis, mass calcification, and some infiltration into the surrounding tissue. Pathological examination suggests tumor cell pleomorphism, more frequent pathological division, high mitotic index, abundant interstitial sinusoid, necrosis, tumor infiltration, vascular invasion, and Weiss score of≥3 points. Lactate acid dehydrogenase and cortisol levels; Weiss score; and tumor size, staging, and resection rate were not statistically different among prognostic factors. Conclusions: For pediatric patients with adrenal masses, the survival rate should be improved by combining the clinical manifestation, laboratory examination, and auxiliary examination for the early diagnosis of adrenocortical carcinoma and to select the appropriate treatment. Combined with pathological results, adrenocortical carcinoma can be well differentiated from other adrenal masses.

Objective:To evaluate the effect of propofol on right ventricular hypertrophy induced by pulmonary arterial hypertension (PAH) in rats.Methods:Twenty-two clean-grade healthy adult male Wistar rats, weighing 250-280 g, were divided into 3 groups using a random number table method: control group (group C, n=8), PAH group (group PH, n=6) and propofol group (group P, n=8). In PH and P groups, monocrotaline 60 mg/kg was injected intraperitoneally to establish the model of PAH, while the equal volume of normal saline was administered in group C. Propofol 100 mg/kg was injected intraperitoneally twice a week for 6 consecutive weeks starting from 2 weeks after establishment of the model in group P. The weight of rats was measured before establishment of model and after administration, and the weight difference (△BW=weight after administration-weight before administration) was calculated.At the end of administration, the right ventricular end-diastolic dimension (RVEDD), right ventricular wall thickness in diastole (RVWTd), intraventricular septum in diastole (IVSd), left ventricular posterior wall in diastole (LVPWd) and maximal velocity of pulmonic valve (PV) were measured using cardiac ultrasound.The animals were then sacrificed, and the lungs and hearts were removed for examination of the pathological changes (after haematoxylin and eosin staining) and for determination of the degree of myocardial fibrosis in right ventricular (by Masson staining), expression of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in lung tissues and myocardial tissues of the right ventricle (by immunohistochemistry). Results:Compared with group C, △BW and PV were significantly decreased, RVWTd, IVTd and RVEDD were increased, the thickness of the wall of pulmonary arterioles and myocardial cell penetration area in right ventricular were increased, the expression of IL-6 and TNF-α in lung tissues and myocardial tissues of right ventricle was up-regulated ( P<0.05 or 0.01), inflammatory cell infiltration and structural disorders were found in lung tissues, and intercellular spaces were widened, and the myocardial tissue was extensively fibrotic in group PH.Compared with group PH, △BW and PV were significantly increased, RVWTd, IVTd and RVEDD were decreased, the thickness of the wall of pulmonary arterioles and myocardial cell penetration area (the area of cardiomyocytes in which the nucleus located in the middle) were decreased, the expression of IL-6 and TNF-α in lung tissues and myocardial tissues of right ventricle was down-regulated ( P<0.05 or 0.01), inflammatory cell infiltration, structural disorders, intercellular spaces and degree of fibrosis were improved in group P. Conclusion:Propofol can alleviate right ventricular hypertrophy induced by PAH, and the mechanism is probably related to reduction of inflammatory responses in rats.

Objective:According to the international cooperation project of Peking University Cancer Hospital on human genetic resource management practices, combined with the development direction of human genetic resource management laws and regulations, and propose reference suggestions for medical institutions to strengthen human genetic resource management.Methods:Sort out the projects that Peking University Cancer Hospital obtained international cooperation approval on the government platform of the Ministry of Science and Technology from July 2019 to June 2023, analyze the current situation of human genetic resource management in the hospital, summarize the challenges brought by the implementation of new regulations on human genetic resource management in medical institutions, and propose corresponding suggestions.Results:A total of 1276 international cooperation projects on human genetic resources have been approved, including 345 initial declarations and 931 change declarations. Involving 453 studies, including 286 clinical trials of drugs or devices on the market, accounting for 63.13%, and 100 clinical trials of Phase I drugs, accounting for 34.97% of the market studies. On average, there are 3.14 changes per project for listed research, and 1.56 changes per project for non listed research.Conclusions:Regulations on the Management of Human Genetic Resources ( short for Rules) limit the management scope of international cooperation projects involving human genetic resources and delegate management authority to medical institutions. Adjusting the scope of application for international cooperative clinical trial filing may result in some administrative approval projects being transferred to filing. The approval process for international cooperative scientific research projects on human genetic resources has been adjusted. Suggest medical institutions to strengthen the management of samples and intellectual property outside the scope of application of Rules.Strengthen the entire process management of international cooperation in scientific research. Pay attention to and timely communicate the dynamics of human genetic resource management.

Objective:To explore the effect of long non-coding RNA (lncRNA) C10orf25 on the proliferation and invasion ability of prostate cancer cells and the possible role of miRNA-671-5p (miR-671-5p).Methods:Data from the Gene expression omnibus (GEO) database (data updated in January 2023) were used to analyze the differences in the expression levels of C10orf25 in 137 cases of prostate cancer tissues and paracancerous tissues. Prostate cancer C4-2B, DU-145, 22Rv1, PC-3, LNCaP cell lines and immortalized prostate epithelial RWPE-1 cell lines were selected, and then real-time quantitative fluorescence polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of C10orf25 in cell lines. The 22Rv1 cells with the lowest relative expression level of C10orf25 were selected and divided into the control group (transfected with negative plasmid) and the C10orf25 group (transfected with C10orf25 plasmid); the CCK-8 method was used to detect the proliferation activity of 22Rv1 cells in both groups at day 1, 2, 3, 4, 5 (expressed as absorbance value); the Transwell method was used to detect the invasion ability of 22Rv1 cells. Linc2GO software was used to predict miR-671-5p with binding sites for C10orf25. Dual luciferase reporter gene assay was used to verify the targeting relationship between C10orf25 and miR-671-5p. qRT-PCR was used to detect the relative expression levels of C10orf25 and miR-671-5p. Western blot was used to detect the expression of proteins related to the NF-κB signaling pathway of 22Rv1 cells in the both groups.Results:In the GEO database, the relative expression level of C10orf25 in prostate cancer tissues was lower than that in paracancerous tissues ( P < 0.01). The relative expression levels of C10orf25 in immortalized prostate epithelial cell line RWPE-1 and prostate cancer cell lines C4-2B, DU-145, 22Rv1, PC-3, and LNCaP were 1.00±0.05, 0.63±0.04, 0.42±0.03, 0.18±0.04, 0.81±0.02, 0.50±0.07, and the difference was statistically significant ( F = 43.29, P < 0.05). The proliferation ability of 22Rv1 cells in C10orf25 group was lower than that in the control group from the second day, and the differences were statistically significant (all P < 0.05). The number of invasive cells in the control group and C10orf25 group were (97±11) and (36±9), respectively, and the difference was statistically significant ( t = 4.15, P < 0.01). Linc2GO software prediction results showed that C10orf25 had a binding site for miR-671-5p. The dual luciferase reporter gene assay showed that the relative luciferase activity of miR-671-5p and C10orf25 wild plasmid co-transfecting 22Rv1 cells was lower than that of miR-NC and C10orf25 wild plasmid co-transfecting 22Rv1 cells, and the difference was statistically significant ( P < 0.01); when miR-671-5p or miR-NC was co-transfected with C10orf25 mutant plasmid, the difference in the luciferase activity of 22Rv1 cells was not statistically significant ( P > 0.05). The relative expression levels of miR-671-5p in 22Rv1 cells were 7.33±0.99 and 0.98±0.16, respectively in the control group and C10orf25 group, and the difference was statistically significant ( t = 6.32, P < 0.01). The results of Western blot showed that the expression levels of NF-κB signaling pathway protein p50, matrix metalloproteinase 9, c-myc, and vascular endothelial growth factor protein in 22Rv1 cells in C10orf25 group were lower than those in the control group. Conclusions:The overexpression of C10orf25 may inhibit the proliferation and invasion of prostate cancer cells through the miR-671-5p-NF-κB axis.