Enhancing Vibrio vulnificus infection diagnosis for negative culture patients with metagenomic next-generation sequencing.

Objective: To evaluate the diagnostic value of metagenomic next-generation sequencing (mNGS) in Vibrio vulnificus (V. vulnificus) infection. Methods: A retrospective analysis of patients with V. vulnificus infection at the Fifth Affiliated Hospital of Sun Yat-Sen University from January 1, 2020 to April 23, 2023 was conducted. 14 enrolled patients were diagnosed by culture or mNGS. The corresponding medical records were reviewed, and the clinical data analyzed included demographics, epidemiology laboratory findings, physical examination, symptoms at presentation, antibiotic and surgical treatment, and outcome. Results: In this study, 78.6% (11/14) patients had a history of marine trauma (including fish stab, shrimp stab, crab splints and fish hook wounds), 7.1% (1/14) had eaten seafood, and the remaining 14.3% (2/14) had no definite cause. Isolation of V. vulnificus from clinical samples including blood, tissue, fester and secreta. 9 cases were positive for culture, 5 cases were detected synchronously by mNGS and got positive for V. vulnificus. 85.7% (12/14) cases accepted surgical treatment, with 1 patient suffering finger amputated. 14 enrolled patients received appropriate antibiotic therapy, and all of them had recovered and discharged. 9 strains V. vulnificus isolated in this study were sensitive to most beta-lactam antibiotics, aminoglycosides, quinolones, etc. Conclusion: Vibrio vulnificus infection is a common water-exposed disease in Zhuhai, which requires identification of a number of pathogens. Of severe infections with unknown pathogen, mNGS can be used simultaneously, and the potential to detect multiple pathogens is of great help in guiding treatment.

Overall survival comparison between pediatric and adult Ewing sarcoma of bone and adult nomogram construction: a large population-based analysis.

Background: Ewing sarcoma (ES) is a common primary bone tumor in children. Our study aimed to compare overall survival (OS) between pediatric and adult bone ES patients, identify independent prognostic factors and develop a nomogram for predicting OS in adult patients with ES of bone. Methods: We retrospectively analyzed data for the 2004-2015 period from the Surveillance, Epidemiology, and End Results (SEER) database. To guarantee well-balanced characteristics between the comparison groups, propensity score matching (PSM) was used. Kaplan-Meier (KM) curves were used to compare OS between pediatric and adult patients with ES of bone. Univariate and multivariate Cox regression analyses were used to screen independent prognostic factors for ES of bone, and a prognostic nomogram was constructed by using the factors identified. The prediction accuracy and clinical benefit were evaluated using receiver operating characteristic (ROC) curves, areas under the curves (AUCs), calibration curves, and decision curve analysis (DCA). Results: Our results showed that adult ES patients had lower OS than younger ES patients. Age, surgery, chemotherapy, and TNM stage were independent risk factors for bone ES in adults and were used to develop a nomogram. AUCs for 3-, 5-, and 10-year OS were 76.4 (67.5, 85.3), 77.3 (68.6, 85.9) and 76.6 (68.6, 84.5), respectively. Calibration curves and DCA results indicated excellent performance for our nomogram. Conclusion: We found that ES pediatric patients have better OS than adult ES patients, and we constructed a practical nomogram to predict the 3-, 5- and 10-year OS of adult patients with ES of bone based on independent prognostic factors (age, surgery, chemotherapy, T stage, N stage and M stage).

Socioeconomic status influences on bone mineral density in American men: findings from NHANES 2011-2020.

The association between socioeconomic status (SES) and bone mineral density (BMD) in men remains controversial. We showed that SES was positively associated with BMD in American men. Confounding factors like race/ethnicity and age could affect the association. INTRODUCTION: Based on the data from the National Health and Nutrition Examination Survey (NHANES), 2011-2020, this article aims to investigate the association of SES (poverty income ratio (PIR) and education level) with the BMD in American men. METHODS: We evaluated the association of SES with BMD in 4446 men aged ≥ 20 years (mean age, 41.0 ± 13.4 years) from the NHANES 2011-2020. BMD was measured by dual-energy X-ray absorptiometry (DXA) at the lumbar spine. We used multivariate linear regression models to examine the relationship between SES and total spine BMD, adjusted for a large range of confounding factors. RESULTS: Compared with other PIR quarters, individuals in the highest quarter of PIR were more likely to be older and white and had fewer smoking or drinking behaviors. After adjusting for race/ethnicity, age, drinking and smoking behavior, body mass index (BMI), total protein, serum calcium, serum uric acid, cholesterol, serum phosphorus, and blood urea nitrogen, PIR was positively correlated with total spine BMD (ß = 0.004 95% CI: 0.001-0.007, P = 0.006). Individuals with the highest degree (college degree or above) had a 0.057 g/cm2 greater BMD than that of the lowest degree (less than 9th grade) (ß = 0.057 95% CI: 0.037-0.077, P < 0.001). CONCLUSIONS: Our study indicates that SES was positively associated with the lumbar BMD among American men. Clinicians, healthcare providers, and policymakers should consider the unequal SES of men when implementing osteoporosis prevention and treatment strategies.

Upregulated periostin promotes angiogenesis in keloids through activation of the ERK 1/2 and focal adhesion kinase pathways, as well as the upregulated expression of VEGF and angiopoietin­1.

Periostin, a secreted extracellular matrix protein, is highly expressed in wound healing and in various types of human cancer and is involved in angiogenesis. Keloids, considered dermal benign tumors, are granulomatous lesions characterized by capillary proliferation. However, the underlying regulatory mechanism of angiogenesis in keloids remains to be elucidated. The present study aimed to examine the effect of periostin on angiogenesis in keloids. The expression of periostin was upregulated and the vessel density was higher in human keloids compared with normal tissue, observed following staining with CD31 and CD105. Periostin demonstrated a markedly positive correlation with blood vessel density, which was assessed using CD31 staining (r=0.711; P<0.01) and a weak correlation was observed using CD105 staining (r=0.251; P<0.01). Conditioned medium from keloid fibroblasts (KFs) promoted the migration and tube formation of human umbilical vein endothelial cells (HUVECs) compared with normal fibroblasts and this effect may have been abrogated by the short hairpin RNA knockdown of periostin. Treatment with recombinant human periostin promoted the migration and tube formation of HUVECs by activating the extracellular signal­regulated kinase 1/2 and focal adhesion kinase signaling pathway. In addition, periostin increased the secretion of vascular endothelial growth factor and angiopoietin­1 in the KFs. In conclusion, these data suggested that upregulation in the level of periostin may promote angiogenesis directly and indirectly in keloids and may be a key factor in keloid development. Periostin may, therefore, be a promising therapeutic target in the treatment of keloids and other angioproliferative diseases.

Increased periostin expression affects the proliferation, collagen synthesis, migration and invasion of keloid fibroblasts under hypoxic conditions.

Periostin, a secreted extracellular matrix protein, is involved in the wound healing and pathological process of various human cancers. Keloid scars are fibroproliferative tumor-like lesions and develop under local hypoxia. Using suppression subtractive hybridization, in a previous study, we found that periostin is overexpressed in keloids compared with hypertrophic scars. However, little is known about the regulation and function of periostin in keloids. In this study, we examined the effects of periostin on the bioactivity of keloid fibroblasts (KFs) in order to determine whether periostin is involved in hypoxia-stimulated keloid pathogenesis by measuring the expression levels of periostin in KFs cultured under hypoxic conditions. We also investigated the association between periostin and hypoxia-inducible factor-1α (HIF-1α). The mRNA, intracellular protein and secreted protein level of periostin was examined by RT-PCR (and quantitative PCR), western blot analysis and enzyme-linked immunosorbent assay (ELISA), respectively. We also used shRNA targeting periostin to knockdown its expression in the KFs. We report that hypoxia (2% O(2)) upregulates both HIF-1α and periostin expression in KFs. In addition, hypoxia-upregulated periostin expression was regulated by HIF-1α. The inhibition of periostin by short hairpin RNA decreased the hypoxia-stimulated proliferation, collagen synthesis, migration and invasion of KFs and altered the cell cycle, but did not affect apoptosis; treatment with recombinant human periostin protein reversed these effects. Periostin also activated the αvß3 integrin-PI3K/Akt pathway in the KFs. These findings suggest that hypoxia initiates hyperplasia of KFs and increases periostin expression under hypoxic conditions; periostin is involved in the pathogenesis of keloids, which indicates that periostin may be a novel therapeutic target for keloids and other fibroproliferative disorders.

[Variations in expressions of periostin and related factors in early stage of wound healing and scar remodeling in rats].

OBJECTIVE: To investigate the expressions of periostin (PN), angiopoietin-1 (Ang-1), vascular epithelial growth factor (VEGF) and fetal liver kinase-1 (Flk-1) during the processes of scar formation and modulation in rat cutaneous wounds and probe into their roles in wound healing and scaring. METHODS: Eighty-two male Sprague-Dawley (SD) rats were randomly divided into 10 groups with 8-9 rats in each group. Two 2 cm×2 cm full-thickness excisional wounds in the back were created in each rat. The wound surface was observed, and the healing area was measured. The pathological change was observed after hematoxylin and eosin (HE) staining. The expressions of PN, Ang-1, VEGF and Flk-1 in wound surface scar at 4-8 weeks were determined with immunohistochemistry. The expressions of PN, Ang-1 and VEGF were determined by Western blotting. The normal skin was served as control. RESULTS: HE staining showed that the wound surface tissue had healed with epithelization at 4-8 weeks. Immunohistochemistry results showed that there was no significant difference in Flk-1 expression between wound surface tissue and normal skin. The PN expression (A value/µm(2)) in wound surface tissue was significantly lower than that in normal skin at 5 weeks (2.43±0.44 vs. 4.24±0.50, P<0.05), and the expression of Ang-1 and VEGF (A value/µm(2)) at 4, 5, 6, 8 weeks was significantly lower than that in normal skin (Ang-1: 3.51±0.93, 3.10±0.57, 2.77±0.59, 2.77±1.26 vs. 4.89±0.48; VEGF: 1.76±0.68, 1.75±0.49, 1.99±0.42, 1.94±0.86 vs. 4.86±1.63, all P<0.05). In wound surface scar, PN and Flk-1 positive signal was found in cell, and the Ang-1 and VEGF positive signal in extracellular matrix. Western blotting data demonstrated that the expressions of PN, Ang-1 and VEGF peaked at the 10th day after excision with increases to 7.90-22.56 folds compared with normal skin (PN: 2.45±1.51 vs. 0.31±0.19, Ang-1: 18.43±15.20 vs. 1.53±1.42, VEGF: 6.09±4.66 vs. 0.27±0.13, P<0.05 or P<0.01), and then followed with a decrease. CONCLUSIONS: PN, Ang-1, VEGF and Flk-1 are transiently overexpressed in early stage of full-thickness cutaneous wound healing in rats. Their expressions vary in wounds and scars. They participate in the healing of full-thickness cutaneous wounds together and may be essential for the proliferation stage during wound healing.

[The expressions of periostin and the related factors during healing process of full-thickness cutaneous wound in rat].

OBJECTIVE: To observe the expression of osteoblast-specific factor 2 (periostin, PN), angiopoietin-1 (Ang-1), vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 [VEGFR-2/fetal liver kinase-1 (FLK-1)] in wound surface and its peripheral skin, and their effects on wound healing in rats. METHODS: Forty-eight Sprague-Dawley (SD) rats were randomly divided into six groups, with 8 rats in each group. An area of 2 cm×2 cm full-thickness skin was excised on both sides of the back of rats. Specimens from wounds were obtained on 1, 4, 7, 10, 14, 21 days after operation, and histological evaluation and immunohistochemical staining of PN, Ang-1, VEGF and FLK-1 were made to determine their expression levels. Normal skin specimens were obtained as normal controls. RESULTS: The expressions of PN, Ang-1, VEGF and FLK-1 were significantly increased in wound surface after operation. Compared with the skin of normal controls, the expression of PN in the tissues of wound increased by 234.4% on the 1st day, and then increased continuously up to 597.9% on the 7th day (reaching the peak) after operation, followed by a decrease, the increase rate was 280.9% on the 21st day, and still remained at a high level (all P < 0.05). The expression of Ang-1 in the tissue of wound increased by 128.1% on the 1st day and 327.5% on the 4th day (reaching the peak), and then, it was gradually decreased. The increase rate was only 80.5% on the 14th day and it rose slightly later (all P < 0.05). The expression of VEGF in the tissues of wound reached the peak (165.8%) on the 7th day. Then it decreased with a slight fluctuation (all P < 0.05). The expression of FLK-1 in the tissues of wound was increased by 56.1% on the 1st day, and the level remained. It reached the peak by an increase of 70.1% on the 7th day (both P < 0.05). Then, it was lowered after the 10th day (all P > 0.05). CONCLUSIONS: The expressions of PN, Ang-1, VEGF, FLK-1 were obviously increased during healing of skin wound, with different peaking time and expressing rates. The increase in expression of PN showed the longest duration and highest peak value. The PN, Ang-1, VEGF, FLK-1 all play a role in the wound healing process, and PN might play an important role during the healing process of a full-thickness cutaneous wound.

[Study on stabilized hyaluronic acid-based gel of non-animal origin for the correction of nasolabial folds].

OBJECTIVE: To assess the safety and efficacy of hyaluronic acid-based gel of non-animal origin (NASHA gel [Restylane]; Q-Med AB, Uppsala, Sweden)for correcting nasolabial folds in Chinese. METHODS: Patients with moderate or severe nasolabial fold (Wrinkle Severity Rating Scale, WSRS) were recruited to receive NASHA gel injection ( < 1.5 ml). The patients were followed up for 6 months. The efficacy was assessed by physicians and patients, respectively. Adverse events (AEs) were recorded and laboratory tests were performed before and after operation. RESULTS: 86 patients were treated. 6 months after injection, improved esthetic results was assessed by patients and physicians independently. 52 AEs happened in 32 cases (37.2%). Most of them were local injection reaction and minor, which were recovered spontaneously. No systemic reaction was found. CONCLUSIONS: NASHA gel can improve the nasolabial folds. It is very safe and tolerated.

The use of peritoneal tissue mobilised with a novel laparoscopic technique to reconstruct a neovagina.

The use of the peritoneum to construct the new lining of the vagina was described 40 years ago. However, the technique required open laparotomy and extensive dissection of the peritoneum and this technical difficulty and morbidities associated with the procedure reduced the acceptance of this technique. Subsequently, outcomes achievable from any neo-vaginoplastic techniques for a vaginal deformity, regardless of the aetiology, have been problematic due to peritoneal reflection at the pelvic floor, lack of ideal tissues to line a neovagina, and cicatricial contracture. The advent of the endoscope renewed interest in using peritoneal tissues in vaginal reconstruction, and peritoneal tissue mobilisation using forceps to grasp the tissue from below has been described. Here, we use a novel device to push a segment of mobilised peritoneum caudally into the vault. We have used this technique successfully in 12 individuals with congenital vaginal agenesis. Under laparoscopic guidance and from the pelvic cavity, a novel instrument was used to push peritoneal tissue down to the introitus, incised, and sutured to the margin of the skin and mucous membrane to form a new introitus. All the new peritoneal linings survived and the diameter, depth, and moisture of the neovaginas allowed for satisfactory sexual intercourse and did not produce unpleasant odour or excessive secretion. Advantages of this operative technique include its simplicity, the reduced possibility of severe complications in the abdominal cavity, the avoidance of severe morbidity in the perineal region, and the production of a functional, hygienic vagina. It is our preferred and recommended method to reconstruct the vagina.

[Laparoscopically assisted neovaginaplasty using peritoneum as vaginal mucous substitute].

OBJECTIVE: To practice a more atraumatic, physiological and aesthetically valued approach of construction for neovagina. METHODS: Laparoscopically using peritoneum as neovagina lining. RESULTS: From March 2005 to September 2006, this technique was adopted to treat 10 patients whose diagnosis was congenital absence of vagina. The ages of the patients were from 19 to 32. The operation lasted average 2.34 hours. And hospitalization was about 20.5 days. Follow-up ranged from 3 - 12 months. No complication occurred. All of the patients was satisfied with their sexual life. CONCLUSIONS: Laparoscopically assisted neovaginaplasty, in which peritoneum was substituted for vaginal mucous membrane, was a kind of ideal approach of vaginal creation.

[Vaginal reconstruction with modified pudendal-thigh flap].

OBJECTIVE: To increase the success rate of vaginal reconstruction with pudendal-thigh flap, reduce the chance of hemocirculatory disorder and relieve the deformity of greater lip of pudendum and vaginal orifice. METHODS: From June 1998 to May 2004, 15 patients, 2 males with transsexualism and 13 females with vaginal absence, aged 23-31 years old (26.2 on average), received vaginal reconstruction. Arterial perforator was detected at 3 cm laterally away from the mid-point between the vaginal orifice and the anus. The flap of 12.0 cm x 5.5 cm was designed at the lateral of the greater lip of pudendum, with the groin as the axis. After the flap was elevated beneath the deep fascia, the fascial pedicle was partially severed from lateral to medium, keeping the perforator intact into the flap. Then the flap was transferred through the tunnel to the socket. The inner layer and the orifice were formed. RESULTS: All flaps survived, and the contour of the vulva was satisfactory. Among the 15 patients, 14 patients' incisions obtained healing by first intention, and only 1 male with transsexualism was discovered to suffer from rectovaginal fistula at the remote juncture of the two flaps at 7 days after the operation. By taking out all the gauze and cleaning the feces, the fistula healed automatically. All patients were followed up for 6 months-3 years. The reconstructed vagina was about 10 cm deep and 2 fingers wide. One female's vagina was found pilous at 20 months after the operation, and no special treatment was given. The other patients' vaginal inner walls were smooth and complete. Two female patients had the experience of sexual intercourse and felt satisfied after using lubricant. CONCLUSION: The modified pudendal-thigh flap has reliable blood supply and makes the transferring during vaginal reconstruction more convenient and the contour of the vulva more satisfactory.

[In-operation adjusting in inferior pedicle technique reduction mammaplasty].

OBJECTIVE: To overcome inflexible disadvantage in reduction mammaplasty design. METHODS: Preoperation, locating approximately new nipple position and redundant breast skin range. In operation, reshaping unfinished breast shape and locating new nipple-areola position finally in near elective position, breast is reduced using inferior pedicle technique. RESULTS: From August, 1995, 34 cases were performed using this method. After 3 - 18 months' follow-up, the result show that there isn't obvious complication, new breast shape is natural, nipple--areola sense exist. CONCLUSIONS: This design method is simple, flexible, operation is safe, effect is reliable.

[Leg re-contouring by using a new technique of partially removing gastrocnemius].

OBJECTIVE: To evaluate a method to partially remove gastrocnemius for improvement of the contour of the leg. METHODS: Thirteen patients with bulked calf were undergoing the treatment. With a self-designed apparatus, the gastrocnemius was partially removed by placing the device in the muscle through an incision in the popliteal fossa. The follow-ups were carried out for 1-6 months. RESULTS: Thirteen patients were successfully treated by above mentioned technique. The removed amount of muscle was weighted between 11 g and 201 g, averaged 77 g. All of the patients could be able to walk three days after the operation. But, the normal walk had to take 1 month after of the surgery for recovering. The function of the ankle joint was not obviously influenced. CONCLUSION: The above mentioned technique is a safe and effective method for re-contouring the leg.

[The histological study of the capsules formed around the mammary prosthesis].

OBJECTIVE: To observe and analyze the pathohistological characteristics of capsules which formed around the mammary prosthesis with different contents. And to provide the selective basis for ideal and safe prosthesis in clinical practice. METHODS: 20 specimen of the capsules were taken from 20 cases who receive the operation of prothesis removal for different reasons. HE, Masson and Mallory staining were used to analyse the tissue structure and characteristics under the light microscope. RESULTS: The common structure including the collagen fibers accumulation, inflammatory cells infiltration and the capillary hyperplasia were found in all specimen. A layer of squamous epithelium-like cell was detected in some specimen. The specific characteristics were also found in different capsules formed around different prosthesis. In the capsules around vegetable oil prosthesis, there was excessive collagen fiber accumulation, and the capsules were much thicker. In the PVP (polyvinylpyrolidone) prosthesis capsules, there was severe inflammatory cell infiltration, and the number of eosinophilic granulocyte increased obviously. In the silicone gel and saline prosthesis capsule, the collagen fibers were well-arranged and the inflammatory cells were much less. Synovial metaplasia was detected in two cases. CONCLUSION: 1. The capsules form around the prosthesis in all cases after mammary augmentation. 2. There will be synovial metaplasis in some cases, for vegetable oil prosthesis, the collagen over-accumulated which lead the capsules become thicker and harder. So it is not a kind of ideal mammary prothesis. 4. The severe infiltration of the inflammatory cells especially the large quantity of eosinophilic granulocyte indicate the possibility of the delayed hypersensitive reaction mediated by eosinophilic granulocyte. Cautious attitude should be taken during application.