Injectable self-assembled dual-crosslinked alginate/recombinant collagen-based hydrogel for endometrium regeneration.

The disadvantages of mainstream therapies for endometrial injury are difficult to resolve, herein, we suggest an omnibearing improvement strategy by introducing an injectable multifunctional self-assembled dual-crosslinked sodium alginate/recombinant collagen hydrogel. The hydrogel possessed a reversible and dynamic double network based on dynamic covalent bonds and ionic interactions, which also contributed to excellent capability in viscosity and injectability. Moreover, it was also biodegradable with a suitable speed, giving off active ingredients during the degradation process and eventually disappearing completely. In vitro tests exhibited that the hydrogel was biocompatible and able to enhance endometrial stromal cells viability. These features synergistically promoted cell multiplication and maintenance of endometrial hormone homeostasis, which accelerated endometrial matrix regeneration and structural reconstruction after severe injury in vivo. Furthermore, we explored the interrelation between the hydrogel characteristics, endometrial structure, and postoperative uterine recovery, which would benefit deep research on regulation of uterine repair mechanism and optimization of hydrogel materials. The injectable hydrogel could achieve favourable therapeutic efficacy without the need of exogenous hormones or cells, which would be of clinical value in endometrium regeneration.

Isolation and identification of antioxidative peptides from crocodile meat hydrolysates using silica gel chromatography.

Crocodiles are cultured in large numbers in Asia and other places in order to protect wild resources and meet the needs of human life. In this study, crocodile (Crocodylus siamensis) meat proteins were extracted and hydrolyzed into peptides, their antioxidant peptides were isolated and purified by silica gel chromatography and identified by LC/MS. Crocodile meat proteins were optimally extracted with water and hydrolyzed by papain based on the degree of hydrolysis and antioxidant activity. The hydrolysates were fractionated by ultrafiltration into 3 kDa, 3-30 kDa, and ≥ 30 kDa fractions. The 3 kDa fraction showed most antioxidant activity of the hydrolysates. Its active peptides were separated by silica gel column chromatography and purified by silica gel TLC, based on TLC bio-autographic assays of the activity. Four highly active peptides were identified by LC/MS as SSLTIQFVEGQFVDSYDPTIENTFTK, VPPHIY, VAPEEHPVLLTEAPLNPK, and RNGLPGPIGPAG. The identified peptides were synthesized and showed 50% free radical scavenging activities at 1.0 mg/mL, equal or higher to ascorbic acid at 0.5 mg/mL, in both DPPH and ABTS assays. The results indicated that the 3 kDa hydrolyzed peptides of crocodile meat had high antioxidant activity and the active peptides can be effectively separated and purified by silica gel column chromatography and TLC.

One-step approach for full-thickness skin defect reconstruction in rats using minced split-thickness skin grafts with Pelnac overlay.

BACKGROUND: Split-thickness skin grafting is the current gold standard for the treatment of traumatic skin loss. However, for patients with extensive burns, split-thickness skin grafting is limited by donor skin availability. Grafting split-thickness skin minced into micrografts increases the expansion ratio but may reduce wound repair quality. Dermal substitutes such as Pelnac can enhance the healing of full-thickness skin wounds, but their application currently requires two surgeries. The present study investigated whether it is possible to repair full-thickness skin defects and improve wound healing quality in a single surgery using Pelnac as an overlay of minced split-thickness skin grafts in a rat model. METHODS: A full-thickness skin defect model was established using male Sprague-Dawley rats of 10 weeks old. The animals were randomly divided into control and experimental groups in which Vaseline gauze and Pelnac, respectively, were overlaid on minced split-thickness skin grafts to repair the defects. Wound healing rate and quality were compared between the two groups. For better illustration of the quality of wound healing, some results were compared with those obtained for normal skin of rats. RESULTS: We found that using Pelnac as an overlay for minced split-thickness skin grafts accelerated wound closure and stimulated cell proliferation and tissue angiogenesis. In addition, this approach enhanced collagen synthesis and increased the formation of basement membrane and dermis as well as the expression of growth factors related to wound healing while reducing scar formation. CONCLUSIONS: Using minced split-thickness skin grafts overlaid with Pelnac enables the reconstruction of full-thickness skin defects in a single step and can increase the healing rate while improving the quality of wound healing.

Simultaneous extraction and separation of oil, proteins, and glucosinolates from Moringa oleifera seeds.

Moringa oleifera is a worldwide cultivated edible and medicinal plant. Its seeds are rich in oil, proteins, and glucosinolates. A practical method was developed to simultaneously extract and separate the three groups of substances from M. oleifera seeds. Smashed seed material was loaded into columns with petroleum ether: ethanol 8:2 (PE-ethanol) and eluted sequentially with 4.8-fold PE-ethanol to extract oil, and 10.8-fold water to extract proteins and glucosinolates. More than 95% of oil, proteins, and glucosinolates were extracted. The extracts were separated automatically into ether (oil) phase and ethanol aqueous phase. The latter was further separated into proteins and glucosinolates by 70% ethanol precipitation. The main glucosinolate was identified by LC-MS as GLC (4-α-rhamnopyranosyloxy-benzyl glucosinolate). After purification, 22.3 g refined oil, 33.0 g proteins, and 5.5 g purified GLC from 100 g M. oleifera seeds were obtained. This study provides a simple and high-efficient method to utilize M. oleifera seeds.

Simplified recovery of enzymes and nutrients in sweet potato wastewater and preparing health black tea and theaflavins with scrap tea.

The industry discards generous organic wastewater in sweet potato starch factory and scrap tea in tea production. A simplified procedure to recover all biochemicals from the wastewater of sweet potato starch factory and use them to make health black tea and theaflavins from scrap green tea was developed. The sweet potato wastewater was sequentially treated by isoelectric precipitation, ultrafiltration and nanofiltration to recover polyphenol oxidase (PPO), ß-amylase, and small molecular fractions, respectively. The PPO fraction can effectively transform green tea extracts into black tea with high content of theaflavins through the optimized fed-batch feeding fermentation. The PPO transformed black tea with sporamins can be used to make health black tea, or make theaflavins by fractionation with ethyl acetate. This work provides a resource- and environment-friendly approach for economically utilizing the sweet potato wastewater and the scrap tea, and making biochemical, nutrient and health products.

Segmental ureteroileal conduit resection for the treatment of distal upper urinary tract recurrence of bladder cancer following cystectomy.

Segmental ureterectomy is less invasive than radical nephroureterectomy and results in nephron preservation and satisfactory tumor control. This study was to determine the feasibility of segmental ureteroileal conduit resection (SUICR) for patients with distal upper urinary tract recurrence of bladder cancer following radical cystectomy. Four patients with high-grade distal upper urinary tract recurrence underwent SUICR 15-108 months after radical cystectomy. The surgical technique details of SUICR, operative results, and follow-up outcomes are reported. The median operation time was 280 min, and estimated blood loss was less than 100 mL. One patient suffered from ileus 5 days after surgery and was managed conservatively. Histopathologic evaluation showed high-grade stages pTa-pT1 diseases for these patients, and ureteral margins were all negative. No patient suffered from tumor recurrence, with a median follow-up of 39 months. SUICR preserved the ipsilateral renal unit and conformed to oncological principles during surgery. The oncological outcome was satisfactory for these properly selected patients. This technique provides a valid alternative to nephroureterectomy for patients with imperative indications and high-grade upper urinary tract recurrence of bladder cancer following radical cystectomy.

Surgical treatment of calcaneal fractures of Sanders type II and III by a minimally invasive technique using a locking plate.

The aim of the present study was to investigate the outcomes of surgical treatment of calcaneal fractures of Sanders type II and III using a minimally invasive technique and a locking plate. We reviewed 33 feet in 33 consecutive patients with Sanders type II and III calcaneal fractures who had undergone a minimally invasive technique using percutaneous reduction and locking plates. All operations were performed by the same surgeons. The postoperative evaluation included radiographs, determination of restoration of Böhler's angle and Gissane's angle, and administration of the American Orthopaedic Foot and Ankle Society ankle-hind foot scale, Maryland Foot Score, and visual analog scale of pain. The mean visual analog scale score was 1.6 ± 1.4 when radiographic fracture healing was observed. The median functional score of the 33 patients (33 feet) reached 82 (interquartile range 80 to 99) at the last follow-up evaluation according to the American Orthopaedic Foot and Ankle Society ankle-hind foot scale and 89 (interquartile range 80 to 99) according to Maryland Foot Score. All cases achieved restoration of a normal Böhler's angle and Gissane's angle. Postoperative superficial infections occurred in 2 patients, subtalar arthritis developed in 2, and no soft tissue necrosis was observed. For Sanders type II and III fractures of the calcaneus bone, treatment with a minimally invasive technique combining percutaneous reduction and locking plate fixation provided satisfactory clinical results, with a lower incidence of complications. However, longer term studies with a larger sample size and more randomized controlled trials are required to define the superiority of our minimally invasive technique compared with conventional surgical treatment of calcaneal fractures.

Laparoscopic ureteral reimplantation underneath broad ligament tunnel for female vesicoureteral stenosis: a technical innovation.

INTRODUCTION: In order to anatomically reconstruct ureteral stenosis, we present a novel technique for laparoscopic ureteral reimplantation. PATIENTS AND METHODS: Three young females, who were diagnosed with hydroureteronephrosis caused by congenital vesicoureteral junction obstruction, were treated by laparoscopic ureteral reimplantation with a tunnel underneath the broad ligament. RESULTS: Surgery was performed successfully without conversion to open surgery. No major intra- or postoperative complications occurred. Postoperative follow-up was 38, 33 and 26 months, respectively. The operative time was between 220 and 260 min. The mean estimated blood loss was less than 20 ml. Subsequent imaging performed 3 months after surgery revealed relief of hydroureteronephrosis for all patients. The patients all gave birth to healthy neonates and showed normal urinary tract sonogram and urine analysis during the gestation period. CONCLUSION: Laparoscopic ureteral reimplantation with broad ligament tunnel is safe and effective, allowing for anatomical reconstruction of ureter defects. However, a larger clinical sample and longer follow-up period will be needed.

Screening, separating, and completely recovering polyphenol oxidases and other biochemicals from sweet potato wastewater in starch production.

Polyphenol oxidase (PPO) has multiple functions, and the lack of commercially available enzyme sources limits its widespread application in various industries. An accurate PPO assay was developed by HPLC determination of the substrate oxidation. Resources screening indicated that sweet potato (Ipomoea batatas L.) wastewater in starch production has high PPO activity. A procedure was developed for separately recovering PPO, ß-amylase, sporamins, and small molecular nutrients (SMNs) from sweet potato wastewater. The wastewater was adjusted to pH 3.5 to precipitate PPO, and then adjusted to 50 % acetone to precipitate ß-amylase and further to 80 % acetone to precipitate sporamins. The SMNs were obtained after acetone recovery. Purified powders of 4.3 × 10(5) units of PPO, 4.0 × 10(6) units of ß-amylase, 8.70 g sporamins, and 20.2 g SMNs were obtained from the wastewater of 1 kg sweet potato. More than 50 million tons of sweet potato is used for starch production annually around the world. Through this simple procedure, huge amount of biochemical resources can be recovered from the wastewater, which greatly increases the economic value of the crop and saves the environment.

Closed reduction and internal fixation versus total hip arthroplasty for displaced femoral neck fracture.

OBJECTIVE: To compare the clinical effects between closed reduction and internal fixation (CRIF) and total hip arthroplasty (THA) for displaced femoral neck fracture. METHODS: In this prospective randomized study, 285 patients aged above 65 years with hip fractures (Garden III or IV) were included from January 2001 to December 2005. The cases were randomly allocated to either the CRIF group or THA group. Patients with pathological fractures (bone tumors or metabolic bone disease), preoperative avascular necrosis of the femoral head, osteoarthritis, rheumatoid arthritis, hemiplegia, long-term bed rest and complications affecting hip functions were excluded. RESULTS: During the 5-year follow-up, CRIF group had significantly higher rates of complication in hip joint, general complication and reoperation than THA group (38.3% vs. 12.7%, P<0.01; 45.3% vs. 21.7%, P<0.01; 33.6% vs. 10.2%, P<0.05 respectively). There was no difference in mortality between the two groups. Postoperative function of the hip joint in THA group recovered favorably with higher Harris scores. CONCLUSION: For displaced fractures of the femoral neck in elderly patients, THA can achieve a lower rate of complication and reoperation, as well as better postoperative recovery of hip joint function compared with CRIF.

New and highly efficient column chromatographic extraction and simple purification of camptothecin from Camptotheca acuminata and Nothapodytes pittosporoides.

INTRODUCTION: Camptothecin, a widely used natural anti-cancer drug, is difficult to extract and purify effectively from plants. OBJECTIVE: To develop new and highly efficient extraction and purification methods for analysis and production of camptothecin from leaves and fruits of Camptotheca acuminata and Nothapodytes pittosporoides roots. METHODS: Dried materials were loaded in empty columns with fivefold 60% ethanol for leaves or 70% ethanol for fruits of C. acumnata, and sixfold 70% ethanol for N. pittosporoides roots. The columns were eluted with the same solvents at room temperature. Eluent was collected as extraction solution. Extraction solution from leaves and fruits of C. acuminata was vacuum-evaporated to remove ethanol, precipitated at pH 8.0 to remove alkaline insolubles and fractionated with chloroform at pH 3.0, which yields a crude product with 70% purity. Extraction solution from N. pittosporoides roots was concentrated to 1/10 volume and precipitated at pH 3.0, which yields a crude product with 60% purity. All crude products were purified by crystallisation. All steps were monitored by HPLC. RESULTS: Camptothecin was extracted from the three plant materials at a 98% rate with 15- or 18-fold solvent for content analysis, or at a 97% rate with five- or sixfold solvent for production. All crude products were purified to 98%. The overall recovery rates of camptothecin from plant materials to purified products reached 92% or higher. CONCLUSION: The new procedures are simple and highly efficient, and have multiple advantages for quantitative analysis and large production of camptothecin from plants.

[Comparison of effectiveness between minimally invasive cannulated screw and open reduction and plate fixation in treatment of humeral greater tuberosity fracture].

OBJECTIVE: To compare the effectiveness between minimally invasive cannulated screw and open reduction and plate fixation in treatment of humeral greater tuberosity fracture by a prospective case-control study. METHODS: Between January 2008 and January 2011, 49 cases of humeral greater tuberosity fractures were treated with minimally invasive cannulated screw in 25 cases (trial group), and with open reduction and plate fixation in 24 cases (control group). There was no significant difference in gender, age, injury cause, disease duration, fracture displacement, injury side, and complications between 2 groups (P > 0.05). The length of incision, operation time, intraoperative blood loss, and hospitalization days were recorded. According to Neer grading system, the effectiveness was evaluated; fracture healing was observed by X-ray films. RESULTS: The trial group had smaller incision, shorter operation time, less blood loss, and shorter hospitalization days than the control group, showing significant differences (P < 0.01). Superficial infection occurred in 2 cases of the control group, and were cured after symptomatic treatment; primary healing of incision was obtained in the others of 2 groups. All patients were followed up 1-4 years (mean, 2.3 years). The fracture healing time was (7.0 + 2.3) weeks in the trial group, and was (7.8 +/- 2.1) weeks in the control group, showing no significant difference (t=1.24, P=0.22). No heterotopic ossification or loosening and breakage of internal fixation occurred during follow-up. The shoulder function Neer score of the trial group (86.3 +/- 2.8) was significantly higher than that of the control group (80.1 +/- 2.1) (t=6.37, P=0.00). The results were excellent in 14 cases, good in 8 cases, fair in 2 cases, and poor in 1 case with an excellent and good rate of 88.0% in the trial group; the results were excellent in 12 cases, good in 7 cases, fair in 2 cases, and poor in 3 cases with an excellent and good rate of 79.2% in the control group; and difference had no statistical significance (Z=0.83, P=0.41). CONCLUSION: Compared with open reduction and plate fixation, minimally invasive cannulated screw for greater tuberosity fracture has the advantages of simple operation, less trauma, less intraoperative blood loss, and good shoulder function recovery.

Viral therapy for pancreatic cancer: tackle the bad guys with poison.

Pancreatic cancer is one of the most devastating diseases with very poor prognosis. Only a small proportion is curable by surgical resection, whilst standard chemotherapy for patients with advanced disease has only modest effect with substantial toxicity. Therefore, there is an urgent need for the development of novel therapeutic approaches to improve the patient outcome. Recently the viral therapy is emerging as a novel effective therapeutic approach for cancer with the potential to selectively treat both primary tumor and metastatic lesions. This review provides an overview of the current status of viral treatment for pancreatic cancer, both in the laboratories and in clinical settings.

Anatomic study of popliteus complex of the knee in a Chinese population.

This study explored the anatomic relationship of the popliteus complex (PC) and collected anatomic data of PC in a Chinese population. The anatomic study was performed using 81 formalin-fixed knees. The femoral attachment of the popliteus tendon could be classified into three types with respect to femoral attachment of the lateral collateral ligament. The popliteofibular ligament presented as ligament (87.7%) or fascia (12.3%), originating from the musculotendinous junction of the popliteus muscle or just proximal to it. Given the great variability of the posterolateral structures, reconstructive surgeons should design a method based on individual anatomic features, rather than a fixed reconstructive surgery.

Occurrence, function and potential medicinal applications of the phytohormone abscisic acid in animals and humans.

Abscisic acid (ABA) is an important phytohormone that regulates plant growth, development, dormancy and stress responses. Recently, it was discovered that ABA is produced by a wide range of animals including sponges (Axinella polypoides), hydroids (Eudendrium racemosum), human parasites (Toxoplasma gondii), and by various mammalian tissues and cells (leukocytes, pancreatic cells, and mesenchymal stem cells). ABA is a universal signaling molecule that stimulates diverse functions in animals through a signaling pathway that is remarkably similar to that used by plants; this pathway involves the sequential binding of ABA to a membrane receptor and the activation of ADP-ribose cyclase, which results in the overproduction of the intracellular cyclic ADP-ribose and an increase in intracellular Ca²âº concentrations. ABA stimulates the stress response (heat and light) in animal cells, immune responses in leukocytes, insulin release from pancreatic ß cells, and the expansion of mesenchymal and colon stem cells. ABA also inhibits the growth and induces the differentiation of cancer cells. Unlike some drugs that act as cell killers, ABA, when functioning as a growth regulator, does not have significant toxic side effects on animal cells. Research indicated that ABA is an endogenous immune regulator in animals and has potential medicinal applications for several human diseases. This article summarizes recent advances involving the discovery, signaling pathways and functions of ABA in animals.

Cloning, microbial expression and structure-activity relationship of polyphenol oxidases from Camellia sinensis.

Polyphenol oxidase (PPO) can be used for organic synthesis and degradation of wastes and dyes in industries. Lack of enzyme sources is a major barrier for its application. A PPO gene, with a full length of 1.8kb without introns, was cloned by PCR from genomic DNA of five common cultivars of Camellia sinensis. They had a 98.2-99.9% degree of identity in nucleotides and 94.7-96.1% in amino acids and encoded a polypeptide of 599 amino acids with a signal peptide targeting the chloroplast and three Cu-binding domains. The mature PPO showed high expression and enzyme activity after refolding the inclusion bodies in Escherichia coli BL21 (DE3) using pET30c expression vector, but low expression in Pichia pastoris GS115 using both the secretory and non-secretory vectors pPICZalphaA and pPICZA. The expression of PPO mutants demonstrated that the signal sequences prevented recombinant gene expression in E. coli. PPO activity was not affected by the C-terminus and was slightly inhibited by the CuC domain. Other domains were important for its activity. A 3.1-fold increase in PPO activity over non-recombinant controls was obtained by expressing the PPO fragment without signal sequences and the CuC domain in E. coli BL21 (DE3) using the pET30c vector.

Cloning and expression analysis of an aldehyde oxidase gene in Arachis hygogaea L.

Aldehyde oxidase (AO) plays important role in plant hormone biosynthetic pathways, such as abscisic acid (ABA) and indole-3-acetic acid (IAA). The enzyme catalyzes the last step of the pathways. In this study a full-length cDNA encoding an aldyhyde oxidase was cloned and sequenced from leaves of peanut by RT-PCR, RACE-PCR and genomic DNA walking methods. The full-length cDNA, designated as Arachis hygogaea L. aldehyde oxidase 1 (AhAO1), consists of an open reading frame of 4131 bp, a 326 bp 5' untranslated region and a 128 bp 3' untranslated region including a poly (A) tail of 21 nucleotides. The gene encodes a polypeptide of 1377 amino acids with a calculated molecular weight of 150 kDa and an isoelectric point (pl) of 6.99. Analysis of amino acid sequence of AhAO1 shows that it had 61%, 59% and 55% identity with the AOs from tomato, Arabidopsis and maize, respectively The peanut AO polypeptide contains consensus sequences for iron-sulfur centers and a molybdenum cofactor (MoCo)-binding domain. Semi-quantitative RT-PCR analysis showed that AhAO1 expression was higher in leaves than in roots of peanut.