Metabolism, fibrosis, and apoptosis: The effect of lipids and their derivatives on keloid formation.

Keloids, pathological scars resulting from skin trauma, have traditionally posed significant clinical management challenges due to their persistence and high recurrence rates. Our research elucidates the pivotal roles of lipids and their derivatives in keloid development, driven by underlying mechanisms of abnormal cell proliferation, apoptosis, and extracellular matrix deposition. Key findings suggest that abnormalities in arachidonic acid (AA) synthesis and non-essential fatty acid synthesis are integral to keloid formation. Further, a complex interplay exists between lipid derivatives, notably butyric acid (BA), prostaglandin E2 (PGE2), prostaglandin D2 (PGD2), and the regulation of hyperfibrosis. Additionally, combinations of docosahexaenoic acid (DHA) with BA and 15-deoxy-Δ12,14-Prostaglandin J2 have exhibited pronounced cytotoxic effects. Among sphingolipids, ceramide (Cer) displayed limited pro-apoptotic effects in keloid fibroblasts (KFBs), whereas sphingosine 1-phosphate (S1P) was found to promote keloid hyperfibrosis, with its analogue, FTY720, demonstrating contrasting benefits. Both Vitamin D and hexadecylphosphorylcholine (HePC) showed potential antifibrotic and antiproliferative properties, suggesting their utility in keloid management. While keloids remain a prevalent concern in clinical practice, this study underscores the promising potential of targeting specific lipid molecules for the advancement of keloid therapeutic strategies.

[Trans-Douglas Retzius' space-sparing robot-assisted simple prostatectomy for large-volume benign prostate hyperplasia].

OBJECTIVE: To report the safety and efficacy of trans-Douglas Retzius' space-sparing robot-assisted simple prostatectomy (RSS-RASP) in the treatment of large-volume BPH. METHODS: This retrospective study included 24 cases of large-volume (>80 ml) BPH treated by trans-Douglas RSS-RASP from August 2019 to June 2021. The patients ranged in age from 55 to 80 (mean 68.5) years, with an average body mass index of 25.1 (20.5－34.9) kg/m2 , median prostate volume of 132.4 (85.6－235.7) ml, and preoperative tPSA of 10.8 (0.5－37.9) ng/ml, IPSS of 25 (3－35) and quality of life (QOL) score of 5 (3－8). Before surgery, 12 of the patients received catheterization for urinary retention, 1 underwent cystostomy, 2 were complicated with hydronephrosis, 1 had stones and diverticulum in the bladder, and 14 were excluded from the cases of PCa by prostatic biopsy. The operation time, intraoperative blood loss, hemoglobin level on the first day after surgery, blood transfusion, and intra- and postoperative complications were recorded. The patients were followed up for 3 to 21 months postoperatively. Comparisons were made before and after operation in the IPSS, maximum urinary flow rate (Qmax), postvoid residual volume (PVR), QOL score, IIEF score and Male Sexual Health Questionnaire (MSHQ) score. RESULTS: Trans-Douglas RSS-RASP was successfully completed in all the 24 cases, with a mean operation time of 175 (100－285) min, intraoperative blood loss of 200 (50－800) ml, hemoglobin decrease of 25 (4－57) g/L on the first day after surgery, postoperative drainage tube indwelling of 3 (2－7) d, and urinary catheterization of 12 (4－18) d. Six (25%) of the patients received intraoperative blood transfusion, 1 underwent transurethral electrocoagulation hemostasis 1 month after surgery because of postoperative bleeding, and 1 received transurethral resection of the cicatrical adhesive tissue of the bladder neck 12 months after surgery. No other complications occurred postoperatively. The IPSS (3 ［1－7］), Qmax (19.6 ［9.9－32.1］ ml/s), PVR (0 ［0－34.9］ ml) and QOL score (2 ［0－3］) of the patients were significantly improved after surgery (P < 0.05), but no statistically significant differences were observed in the IIEF (20 ［19－24］) and MSHQ scores (14 ［13－14］) as compared with the baseline (P > 0.05). CONCLUSION: Trans-Douglas RSS-RASP is a safe and effective minimally invasive method for the treatment of large-volume (>80 ml) BPH, which can improve the urinary function of the patient after operation.

Visual upconversion nanoparticle-based immunochromatographic assay for the semi-quantitative detection of sibutramine.

Immunochromatographic assay (ICA) has been used widely for the onsite monitoring of illegal additives due to its simplicity, speed, and low cost. However, a scanner is commonly required for ICA to achieve quantitative results. In this work, we developed a visual semi-quantitative ICA for sibutramine, a banned additive in diet foods, without the need for a scanner for measurement. Monoclonal antibodies specific for sibutramine were raised and conjugated with upconversion nanoparticles (UCNPs) as the luminescent tracer. ICA was developed by employing multiple test lines to achieve the semi-quantitative detection of sibutramine. Based on the optimal conditions, the cutoff levels (limit of quantitation, LOQ) of T1 line, T2 line, T3 line, and T4 line were 0.02 µg/mL, 0.15 µg/mL, 1.0 µg/mL, and 7.5 µg/mL, respectively, in buffer system. The ICA demonstrated a LOQ at 0.2 mg/kg for sibutramine in diet food samples. The assay (including pretreatment) can be finished within 30 min without the aid of other instruments, except a laser pen. No false positive or false negative results were observed. The results indicated that the proposed method was reliable, simple, and rapid for the screening of sibutramine abuse in diet food samples.

[Nerve-sparing robot-assisted laparoscopic radical cystectomy: Clinical application and effect].

OBJECTIVE: To investigate the clinical feasibility and effect of nerve-sparing robot-assisted laparoscopic radical cystectomy (NSRA-LSRC). METHODS: We retrospectively reviewed the clinical data on 12 cases of NSRA-LSRC performed from March 2016 to May 2018. The patients were aged 45 to 65 years old and all potent before surgery, with a mean IIEF-5 score of >17. The surgical procedure involved excision of the bladder and prostate and dissection of the pelvic lymph nodes, with preservation of the bilateral neurovascular bundles, internal accessory pudendal artery and pubic bladder complex. All the patients were advised to take PDE5I postoperatively and followed up for the sexual function with the IIEF-5 scores. RESULTS: Surgical procedures were completed successfully, all with negative surgical margins. Postoperative pathology confirmed invasive high-grade urothelial carcinoma or carcinoma in situ in all the cases, including 11 cases in stage T2N0M0 or below and 1 case in stage T3aN0M0. There were no serious intraoperative or postoperative complications, nor recurrence or metastasis during the follow-up period of 12－36 (20.7 ± 8.0) months. The IIEF-5 scores of the patients at 3, 6 and 12 months after operation were 10.9 ± 6.9, 12.3 ± 6.9 and 14.1 ± 8.0, respectively. At 12 months, satisfactory sexual intercourse was achieved with the help of potency-enhancing medicine in 5 cases (41.7%), penile erection insufficient for sexual intercourse in 3 cases (25%), and no erection in 4 cases (33.3%). CONCLUSIONS: Nerve-sparing robot-assisted laparoscopic radical cystectomy can maximally preserve the sexual function of the patients with urinary bladder carcinoma.

[Retzius-sparing robot-assisted laparoscopic radical prostatectomy for early-stage prostate cancer (with video)].

OBJECTIVE: To investigate the application of Retzius-sparing robot-assisted radical prostatectomy (RS-RARP) in the treatment of early-stage prostate cancer. METHODS: We retrospectively analyzed the clinical data about 10 cases of early-stage prostate cancer treated by RS-RARP with the Da Vinci Robot Surgical System from September to October 2016. RESULTS: All the operations were successfully completed without positive surgical margins. The operation time was 170－250 min (ï¼»196±25ï¼½ min), the intraoperative blood loss was 150－500 ml (ï¼»260±128ï¼½ ml), the postoperative hospital stay was 6－7 days, and the catheterization time was 14 days. Urinary continence occurred after catheter removal in 1 patient and was recovered 1 month later. CONCLUSIONS: RS-RARP is a safe, effective and reliable method for the treatment of prostate cancer and conducive to the early recovery of urinary continence.

Genome-wide copy number variation study in anorectal malformations.

Anorectal malformations (ARMs, congenital obstruction of the anal opening) are among the most common birth defects requiring surgical treatment (2-5/10 000 live-births) and carry significant chronic morbidity. ARMs present either as isolated or as part of the phenotypic spectrum of some chromosomal abnormalities or monogenic syndromes. The etiology is unknown. To assess the genetic contribution to ARMs, we investigated single-nucleotide polymorphisms and copy number variations (CNVs) at genome-wide scale. A total of 363 Han Chinese sporadic ARM patients and 4006 Han Chinese controls were included. Overall, we detected a 1.3-fold significant excess of rare CNVs in patients. Stratification of patients by presence/absence of other congenital anomalies showed that while syndromic ARM patients carried significantly longer rare duplications than controls (P = 0.049), non-syndromic patients were enriched with both rare deletions and duplications when compared with controls (P = 0.00031). Twelve chromosomal aberrations and 114 rare CNVs were observed in patients but not in 868 controls nor 11 943 healthy individuals from the Database of Genomic Variants. Importantly, these aberrations were observed in isolated ARM patients. Gene-based analysis revealed 79 genes interfered by CNVs in patients only. In particular, we identified a de novo DKK4 duplication. DKK4 is a member of the WNT signaling pathway which is involved in the development of the anorectal region. In mice, Wnt disruption results in ARMs. Our data suggest a role for rare CNVs not only in syndromic but also in isolated ARM patients and provide a list of plausible candidate genes for the disorder.

Extended Left Hemihapatectomy with Right Hepatic Artery Reconstruction for Primary Hepatic Neuroendocrine Neoplasm: A Brief Report.

BACKGROUND rimary hepatic neuroendocrine neoplasms (PHNEN) are exceedingly rare tumors with atypical clinical manifestations, accounting for less than 0.5% of all neuroendocrine tumors. Currently, there is a lack of consensus on their management, and guidelines do not recommend postoperative chemotherapy for patients with stage G1/G2 disease after curative resection. We present a case report of PHNEN, outlining its diagnostic challenges, treatment strategy, and clinical outcomes. CASE REPORT A 31-year-old man presented with jaundice and was initially diagnosed with suspected IgG4-related disease, which initially appeared to respond to steroid therapy, but manifested worsening jaundice 4 months after initial treatment. Subsequent evaluation revealed a PHNEN NET G2 with lymph node metastasis and invasion of the right hepatic artery; and involvement of the hepatic duct at the hepatic hilum, primarily the left hepatic duct. The patient underwent extended left hemi-hepatectomy with caudate lobe resection, bile duct resection, and lymphadenectomy, followed by reconstruction of the right hepatic artery. Postoperatively, the patient received adjuvant chemotherapy consisting of capecitabine (1000 mg bid D1-14) and temozolomide (200 mg qn D10-14) for 6 cycles. Currently, the patient remains disease free 43 months after treatment. CONCLUSIONS PHNEN presents diagnostic challenges due to its rarity and lack of specific markers. Surgical resection remains the cornerstone of treatment, with chemotherapy being considered in select cases with high-risk features. Further research is needed to refine treatment approaches and improve outcomes for patients with PHNEN.

Targeting NAD metabolism regulates extracellular adenosine levels to improve the cytotoxicity of CD8+ effector T cells in the tumor microenvironment of gastric cancer.

PURPOSE: Nicotinamide adenine dinucleotide (NAD+) is closely related to the pathogenesis of tumors. However, the effect of NAD+ metabolism of gastric cancer (GC) cells on immune cells remains unexplained. We targeted nicotinamide phosphoribosyltransferase (NAMPT), a rate-limiting enzyme in the NAD+ synthesis salvage pathway, to observe its effect in the immune microenvironment. METHODS: NAMPT of GC cell lines was inhibited by using the small molecule inhibitor (FK866) and short hairpin RNA (shRNA). CCK-8 test and flow cytometry were performed to detect cell viability and apoptosis. Immunofluorescence was used to observe changes in mitochondrial membrane potential (MMP).The transfected GC cells (AGS) and patient-derived organoids (PDOs) were cocultured with activated PBMCs, followed by flow cytometric analysis (FCA) for cytokines and inhibitory marker. The level of NAD and ATP of GC cells (AGS & MKN45) was tested combined with NMN and CD39 inhibitor. RESULTS: Targeting NAD+ by FK866 obviously reduced MMP, which ultimately inhibited proliferation and increased the apoptosis of GC cells. NAMPT silencing reduced intracellular NAD and ATP，further decreased extracellular adenosine. Meawhile, the cytokines of CD8+T cells were significantly increased after cocultured with transfected AGS, and the expression of PD-1 was distinctly decreased. NMN reversed the effect of shNAMPT and enhanced the immunosuppression. Consistent results were obtained by coculturing PBMCs with PDOs. CONCLUSION: Restraining the function of NAMPT resulted in the functional improvement of effector CD8+ T cells by decreasing extracellular adenosine levels and inducing apoptosis of GC cells simultaneously. Therefore, this study demonstrates that NAMPT can be an effective target for gastric cancer immunotherapy.

Detection of PAX3/PAX7-FKHR fusion transcripts in rhabdomyosarcoma and other small round cell tumors by 1-step reverse transcriptase polymerase chain reaction: a novel tool for diagnosis and differentiation.

Rhabdomyosarcoma (RMS) is one of the most frequent soft tissue sarcomas in children. It is sometimes difficult to distinguish it from other small round cell tumors (SRCTs) depending on microscopic observations; although their treatment and prognosis varied widely, the same happens between alveolar RMS (ARMS) and embryonal RMS (ERMS). The role of PAX3/PAX7-FKHR fusion gene has been reported in ARMS but not in ERMS and SRCT. The aim of this study was to explore its value in RMS diagnosis and differentiation. Ninety-eight patients with ARMS (n = 13), ERMS (n = 25), pleomorphic RMS (n = 5), Ewing sarcoma (n = 11), neuroblastoma (n = 18), lymphoma (n = 24), and uncertain SRCT (n = 2) were analyzed. One hundred fifteen RNA samples were extracted from the primary tumor tissue at initial presentation and relapse. One-step reverse transcriptase polymerase chain reaction assays for the PAX3/PAX7-FKHR fusion transcripts were performed. Molecular findings were compared with original histologic diagnoses. PAX3-FKHR fusion transcript was detected in 9 ARMS samples, PAX7-FKHR fusion transcript was detected in 7 ARMS samples, and 2 uncertain SRCTs were detected; none of them were detected in ERMS, Ewing sarcoma, neuroblastoma, and lymphoma. Direct sequencing of PAX3 coding regions revealed a heterozygous mutation A→G (nt1380) at codon 448 (AAT→GAT), resulting in substitution of Asn-448 for Asp. Detection of PAX3/PAX7-FKHR fusion transcripts by 1-step reverse transcriptase polymerase chain reaction is a novel tool for RMS diagnosis and differentiation.

Application of contrast-enhanced ultrasonography after radiofrequency ablation for renal cell carcinoma: is it sufficient for assessment of therapeutic response?

PURPOSE: To assess the efficacy of contrast-enhanced ultrasonography (CEUS) with Sonovue in the evaluation of therapeutic response to radiofrequency ablation (RFA) of renal cell carcinoma (RCC). MATERIALS AND METHODS: In a recent 3 years, 63 patients (mean age, 60 years; range 26-81 years) with 64 RCCs were treated by RFA. The lesions had a diameter between 1.8 and 9.8 cm (average diameter, 3.1 cm). The indications for RFA treatment included chronic renal insufficiency (n = 10), presence of solitary kidney (n =3), bilateral renal carcinoma (BRCC) (n =2), advanced age (n =12), significant medical comorbidity (n =29) or refusal of conventional therapy (n =7). Tumors were treated by laparoscopy-assisted (n =41), open surgical (n =18) or percutaneous US guidance (n =4). Follow-up CEUS and contrast-enhanced CT were performed 1 month after treatment to assess the necrotic area. Technical success was defined as elimination of areas that enhanced at imaging within the entire tumor. RESULTS: On the 1-month CEUS and CT imaging after RFA, 62 of 64 tumors (96.9%) were successfully ablated with one session, and residual tumors were found in two RCCs. One of the two tumors was subjected to additional RFA treatment. We could not obtain a complete ablation in the other tumor of a patient with solitary kidney. The diagnostic concordance between the CEUS and 1-month follow-up CT was 100%. Sixty-one patients survived in the follow-up phase which ranged from 2 to 34 months. One patient with solitary kidney died of systemic disease progression and one patient was lost to follow-up. Of the 61 tumors without residual on both CT and CEUS after RFA, four had suspicious findings of recurrence on follow-up CEUS, and two of them were confirmed by subsequent CT examination. With CT as the reference imaging procedure in the assessment of renal tumor ablation, the sensitivity, specificity, positive predictive value, and negative predictive value of CEUS for detecting recurrence during follow-up were 100%, 96.6%, 50%, and 100%. CONCLUSION: Despite its limitation of false-positive value, CEUS is potentially effective in assessing the therapeutic response to RFA of RCC.

[Radiofrequency ablation-assisted laparoscopic partial nephrectomy without renal hilar vessel clamping: a report of 14 cases].

OBJECTIVE: To evaluate the clinical feasibility of radiofrequency ablation-assisted laparoscopic partial nephrectomy (RF-LCPN) without renal hilar vessel clamping. METHODS: From January 2009 to June 2010, 14 cases with renal tumors were treated with RF-LCPN. The mean tumor size was 3.1 cm (range: 2.2 - 4.0 cm). All cases were staged at T(1a)N(0)M(0). The preoperative serum creatinine (SCr) was (87 ± 36) µmol/L and glomerular filtration rate (GFR) (45 ± 11) ml/min. RESULTS: RF-LCPN was technically successful in all patients (14 tumors). The mean operative time was (152 ± 46) min and the mean blood loss (89 ± 52) ml. All tumor margins were negative. One patient with a brief period of urine leakage from lower pole calix was managed successfully by ureteral stenting and Foley catheter drainage of bladder. The postoperative hospital stay was (5 ± 2) days. The postoperative SCr was (90 ± 38) µmol/L and GFR (41 ± 12) ml/min. There was no statistic post-operative change of SCr and GFR (P > 0.05). All patients completed a minimum follow-up of 6 months (mean: 17 months, range: 6 - 25 months). No evidence of local or port-site recurrence or metastasis was found. CONCLUSION: As an accurate and effective intervention with a relatively low incidence of complications, RF-LCPN may eliminate renal warm ischemia. But its long-term efficacy remains to be further studied.

An efficient and user-friendly method for cytohistological analysis of organoids.

Organoid culture is a recently developed in vitro three-dimensional (3D) cell culture technology. It has wide applications in tissue engineering studies. However, histological analysis of organoid is quite complex and tedious for researchers. This study proposes a user-friendly, affordable and efficient method for making formalin-fixed paraffin embedded (FFPE) organoid blocks and Optimal Cutting Temperature compound (OCT) embedded frozen organoid blocks. This method implements a key pre-embedding step for preparing paraffin embedded organoid blocks, which could concentrate organoid together without damaging or loss of samples. This method could be used to process even a small number of organoids with high efficiency. In addition, with minor modifications, the method is readily applied for OCT embedded organoid blocks. The slides generated were ready for H&E staining, immunohistochemistry staining and immunofluorescent staining. The method described in this study can be easily used for routine histological analysis of organoid, and could be performed in general pathology labs and requires no dedicated equipment and reagent.

[A retrospective study comparing the clinical efficacy of laparoscopic cryoablation and partial nephrectomy for renal cell carcinoma].

OBJECTIVE: To compare the clinical outcomes of laparoscopic cryoablation (LCA) and laparoscopic partial nephrectomy (LPN) in the treatment of renal cell carcinoma (RCC). METHODS: Between April 2005 and March 2009, 47 patients were treated with minimally invasive nephron sparing surgery (LPN or LCA) for RCC. The LCA group included 18 selected primary RCC cases (14 men and 4 women, mean age 63 years). There were 6 tumors located in the left, 11 located in the right and 1 located bilaterally. The maximum diameter of tumors was 1.5 - 5.0 cm (mean: 2.9 cm). The LPN group included 29 renal tumors patients (19 men and 10 women, mean age 61 years). The maximum diameter of tumors in this group was 2.0 - 4.5 cm (mean: 2.8 cm). Changes of hemoglobin (Hb), erythrocyte sedimentation rate (ESR), serum creatinine (SCr) and glomerular filtration rate (GFR) after operations were compared between LCA group and LPN group. The operative time, average intra-operative bleeding volume, postoperative hospital stay and incidence of postoperative complications of the 2 groups were analyzed and compared. RESULTS: The 2 surgical procedures were both successful. There was no significant change of Hb, ESR, SCr and GFR after operations in LCA group and LPN group (P > 0.05). The operative time was (94 ± 29) min and (146 ± 45) min in LCA group and LPN group, respectively. The average estimated blood loss was (37 ± 20) ml and (274 ± 69) ml. The postoperative hospital stay was (4 ± 2) d and (10 ± 2) d. These differences between the 2 groups were significant (P < 0.01). No laparoscopic operative complications were noted in LCA group. Follow-up magnetic resonance imaging (MRI) at 1, 3, and 6 months identified the punched-out, nonenhancing, spontaneously resorbing, renal cryolesion. LCA group had completed a minimum follow-up of 6 months (mean 16, range 6 to 21 months). No evidence of local or port-site recurrence was found, and no patient developed metastatic disease. 3 - 36 months' (mean 20 months) follow-up showed no recurrence of tumors or metastatic disease in LPN group. CONCLUSIONS: LCA for RCC is an accurate and effective intervention with a relatively low incidence of complications, and is superior to LPN in operative time, intraoperative bleeding volume and postoperative recovery.

[Inhibitory effect of TRPV6 silencing on prostate cancer cell line LNCaP in vitro].

OBJECTIVE: To explore the effects of the TRPV6 gene silencing by small interfering RNA (siRNA) on the proliferation, cell cycle and apoptosis of human prostate cancer LNCaP cells. METHODS: We constructed two siRNA sequences (siTRPV6-1 and siTRPV6-2) targeting the TRPV6 gene and then transfected them into LNCaP cells mediated by liposome. The transcription of TRPV6 mRNA was detected by RT-PCR, and the effects of siRNA on the proliferation, cell cycle and apoptosis of the LNCaP cells were determined by MITT and flow cytometry. RESULTS: Both siTRPV6-1 and siTRPV6-2 significantly suppressed the expression of TRPV6 mRNA in the LNCaP cells, and the expression was decreased with the extension of time, by 73 and 77% respectively at 72 h after transcription with siTRPV6-1 and siTRPV6-2 as compared with the blank control group (P < 0.01). The proliferation inhibition rates were the highest (34.53 and 29.32%) at 48 h in comparison with 24 and 72 h (P < 0.05). The number of cells was significantly increased in the GO and G1 phases and decreased in the S phase after siTRPV transfection (P < 0.01). The apoptosis rates of LNCaP cells were 14.45 and 12.73% respectively at 48 h after transfected with siTRPV6-1 and siTRPV6-2, significant higher than in the blank control and negative control groups (P < 0.05). CONCLUSION: TRPV6-targeted siRNA can effectively inhibit the transcription of TRPV6 mRNA, inhibit the proliferation of LNCaP cells, arrest their cycles in the G0 and G1 phases, and induce their apoptosis.

Postmenopausal mild hirsutism and hyperandrogenemia due to ovarian Sertoli-Leydig cell tumor: A case report.

Among several types of ovarian tumors, Sertoli-Leydig cell tumors are considered significantly rare, accounting for less than 1% of all primary ovarian tumors. Hirsutism caused by ovarian tumors accounts for approximately 1% of all cases of hirsutism. We report a case of a woman with a ovarian Sertoli-Leydig cell tumor who presented with hirsutism. A 45-year-old woman (gravida 12, para 2) who experienced menopause when she was 43 years old had excessive hair on her face and lower abdomen since 2 years. Her body mass index was 24.3 kg/m2. She also had hair growth on her upper lip, submandibular area, lower abdomen, vulva, and bilateral tibia (front), and around her breast. She had a Ferriman-Gallwey score of 8. Ultrasound findings revealed a 4.8 × 3.5-cm left adnexal mass. Pelvic computed tomography (CT) findings revealed that her left accessory gland had a low-density mass (CT value, 25 Hu). Her serum testosterone level was 15.80 nmol/l. The patient underwent a laparoscopic left adnexectomy. Subsequently, she was diagnosed with ovarian Sertoli-Leydig cell tumor by immunohistochemical staining. A week after surgery, her serum testosterone level decreased from 15.80 nmol/l to 1.03 nmol/L. Her hirsutism almost completely disappeared 3 months after surgery. It is vitally important to establish the final diagnosis according to the clinical manifestations and laboratory values in addition to imaging studies and laparoscopic examination of a rare coexistence of hirsutism and hyperandrogenemia in a postmenopausal woman based on ovarian pathology.

Comparison of changes in body composition during puberty development of obese and normal-weight children in China.

OBJECTIVE: To compare the changes in body composition, including fat mass index (FMI) and fat free mass index (FFMI) during puberty development of obese and normal-weight children in China, and to explore the effect of age and gender on body composition. METHODS: A total of 356 children at the age of 7-15 years were enrolled in this study. Body composition of 10 normal-weight and obese children in each age group was measured by dual-energy x-ray absorptiometry (DEXA). FFMI and FMI were calculated according to the following formula: FFMI (kg x m(-2)) = FFM(kg) / height2 (m2) and FMI (kg x m(-2)) = FM (kg) / height2 (m2). RESULTS: The fat mass and fat free mass of obese children were significantly higher than those of normal-weight children (P < 0.05). The FMI and FFMI of obese children increased significantly with age and were higher than those of the same sex, gender, and age normal-weight children (P < 0.05). CONCLUSION: The levels of fat mass, fat free mass, FMI, and FFMI are different in obese and normal-weight children, and gender effects are significant in boys having higher levels of these indicators than in girls. FFMI and FMI can be used as monitoring indexes in weight control of obese children.

Efficacy of Sox10 Promoter Methylation in the Diagnosis of Intestinal Neuronal Dysplasia From the Peripheral Blood.

OBJECTIVES: Intestinal neuronal dysplasia (IND) is a common malformation of the enteric nervous system. Diagnosis requires a full-thickness colonic specimen and an experienced pathologist, emphasizing the need for noninvasive analytical methods. Recently, the methylation level of the Sox10 promoter has been found to be critical for enteric nervous system development. However, whether it can be used for diagnostic purposes in IND is unclear. METHODS: Blood and colon specimens were collected from 32 patients with IND, 60 patients with Hirschsprung disease (HD), and 60 controls. Sox10 promoter methylation in the blood and the Sox10 expression level in the colon were determined, and their correlation was analyzed. The diagnostic efficacy of blood Sox10 promoter methylation was analyzed by receiver operating characteristic curve. RESULTS: The blood level of Sox10 promoter methylation at the 32nd locus was 100% (90%-100%; 95% confidence interval [CI], 92.29%-96.37%) in control, 90% (80%-90%; 95% CI, 82.84%-87.83%) in HD, and 60% (50%-80%; 95% CI, 57.12%-69.76%) in IND specimens. Sox10 promoter methylation in the peripheral blood was negatively correlated with Sox10 expression in the colon, which was low in control, moderate in HD, and high in IND specimens (r = -0.89). The area under the curve of Sox10 promoter methylation in the diagnosis of IND was 0.94 (95% CI, 0.874-1.000, P = 0.000), with a cutoff value of 85% (sensitivity, 90.6%; specificity, 95.0%). By applying a cutoff value of 65%, promoter methylation was more indicative of IND than HD. DISCUSSION: The analysis of Sox10 promoter methylation in the peripheral blood can be used as a noninvasive method for IND diagnosis.

Association of the common genetic variant upstream of INSIG2 gene with obesity related phenotypes in Chinese children and adolescents.

OBJECTIVE: To study the association between the rs7566605 variant of INSIG2 and obesity-related phenotypes in Chinese children and adolescents. METHODS: The study sample consisted of two independent cohorts of Chinese children and adolescents. Anthropometric indices, lipids, blood pressure, fasting glucose, insulin and percentage of fat mass were determined. PCR with restriction fragment length polymorphism analysis was performed for genotyping the rs7566605 variant. RESULTS: In each of the two independent cohorts, no significant association was observed between rs7566605 and obesity under additive, dominant or recessive model. We also did not detect any difference in the genotype frequency between all the obese children and controls. Furthermore, we did not find evidence of an association between body composition indices and metabolic phenotypes in all children. However, the triglyceride level of CC homozygotes was significantly higher than that of GG+GC genotypes in obese children (P=0.022). Additionally, we observed a non-significant trend of severe obesity in a post-hoc test. CONCLUSION: INSIG2 rs7566605 variant is not associated Chinese childhood obesity in two independent cohorts. Further study is needed to verify the effect of rs7566605 on triglyceride in obese children.

Design and synthesis of immunoconjugates and development of an indirect ELISA for rapid detection of 3, 5-dinitrosalicyclic Acid hydrazide.

In this study novel immunoconjugates were designed, synthesized and then used to develop a rapid, specific and sensitive indirect ELISA method to directly detect residues of 3,5-dinitrosalicyclic acid hydrazide (DNSH), a toxic metabolite of nifursol present in chicken tissues. The hapten DNSHA was first designed and used to covalently couple to BSA to form an immunogen which was immunized to rabbits to produce a polyclonal antibody against DNSH. Furthermore, a novel 3,5-dinitrosalicylic acidovalbumin (DNSA-OVA) immunoconjugate structurally different from DNSHA-OVA was designed and used as a "substructural coating antigen" to improve the sensitivity of an indirect ELISA analysis for a direct DNSH detection. Based on the "substructural coating antigen" concept, an optimized indirect ELISA method was established that exhibited good specificity and high sensitivity for detecting DNSH, with a cross-reactivity of less than 0.1% (excluding the parent compound nifursol), IC(50) of 0.217 nmol/mL and detection limit of 0.018 nmol/mL. Finally, a simple and efficient analysis of DNSH samples in chicken tissues showed that the average recovery rate of the indirect ELISA analysis was 82.3%, with the average coefficient of variation 15.9%. Thus, the developed indirect ELISA method exhibited the potential for a rapid detection of DNSH residues in tissue.

A full-automated magnetic particle-based chemiluminescence immunoassay for rapid detection of cortisol in milk.

A sensitive full-automated micromagnetic particles (MMPs) based competitive chemiluminescent immunoassay (CLIA) was developed to detect cortisol in milk. Polyclonal antibody (pAb) with good specificity against cortisol was produced. The antigen (cortisol-OVA) was labeled with acridinium ester (cortisol-OVA-AE) as signal tracer. During the detection, the free cortisol in sample will compete with cortisol-OVA-AE for binding to pAb. To capture pAb, MMPs conjugated with goat anti-rabbit IgG was added. The whole immunoassay process (exclude sample pretreatment) was performed by automatic chemiluminescence immunoassay instrument, which could consume less test time (within 40 min) and avoid error from manual operation. The method showed a good detection limit of 0.12 ng/mL, a broad linear range from 0.42 to 72.27 ng/mL for cortisol detection, negligible cross-reactivity with related analogues and satisfied recovery (84.3%-102.3%) for spiked milk samples test. Simultaneously, since the results of proposed method had no significant difference with those of LC-MS/MS, the proposed method was confirmed to have a potential applicability for rapidly monitoring cortisol in the food.