Background The select of supporter is critical for the construction of tissue engineering cornea.Many carrier carl be utilized in the construction of tissue engineering cornea,but de-cellular corneal matrix is known to be one of optimal supporters.Objective Present study was to investigate the characteristics of de-cellular corneal matrix of porcine of structure and biocompatibility for rabbit cornea stroma and limbal epithelial ceHs. Methods The porcine cornea was prepared as de-cellular corneal matrix of porcine by the application of detergent enzyme combined process.The corneal epithelial cells,keratocyte and endothelial cells of porcine were removed completely and stored in -20℃ refrigerator after sterilization.The morphology of de-cellular corneal matrix of porcine was examined by hematoxylin-eosin staining under the light microscope.The structure characteristics of de.cellular corneal matrix of porcine under the scan electron microscope,and its physics features were investigated by the evaluation of water content,strength,expansion and transparency.The de-cellular corneal matrix of porcine were implanted to cornea stroma of rabbit and co-cultured with rabbit corneal epithelial cells for 4 weeks in vitro to assess the keracyte compatibility. Results The epithelial cells,keratocyte and endothelial cells of porcine were removed completely by trypsogen digestion.The collagen fibril network and collagen plates paralleled to corneal surface under the light microscope.The water content,strength,expansion。Ratio of light transparency of de-cellular corneal matrix of porcine were similar to normal porcine cornea.After implantation of de.cellular comeal matrix of porcine into rabbits corneal stroma,the edema of tissue was found in one week,and edema disappeared on two weeks and became clear on four weeks after surgery.The de-cellular eorneal matrix attached to rabbit cornea stroma well.No inflammatory eell and new vessel were found after surgery.The co-cultured rabbit corneal epithelial cells differentiated and proliferated on the surfaee of de-cellular corneal matrix and showed positive response for CK3.No statistically significant differences were found in the water content,strength,expansion of de-cellular cornea matrix of porcine among the normal,before dehydration,2 and 4 hours after dehydration cornea matrix(P＞0.05).However,the transparency was much better in the corneal matrix with 2-hour,4-hour dehydration in comparison with non-dehydration one(P＜0.05). Conclusion The structure features of de-cellular cornea matrix of porcine are similar to normal porcine cornea.Good biocompatibility is proved for xenogenesis of rabbit cornea.

Abstract: Objective　To analyze the clinical characteristics and changes of serum IgG, IgM antibodies in patients infected with the SARS-CoV-2 B.1.1.529 (Omicron) variant. Methods　The clinical data of 82 patients with SARS-CoV-2 B.1.1.529 variant was analyzed retrospectively. Based on the presence of pneumonia on chest CT, the patients were divided into pneumonia group and non-pneumonia group. Serum IgG, IgM antibodies were observed at 5 time points T1 (1~<4 d), T2 (4~<8 d), T3 (8~<15 d), T4 (15~<22 d) and T5 (22~<30 d) after admission. Results　Among the 82 patients infected with the SARS-CoV-2 B.1.1.529 variant strain, there were 62 cases of cough, 31 cases of fever, 33 cases of throat discomfort, 5 cases of muscle soreness and 3 cases of diarrhea. The serum IgG antibody levels at 5 time points were 50.22 (142.20) AU/mL, 326.50 (220.63) AU/mL, 368.23 (76.21) AU/mL, 368.65 (79) AU/mL, and 385.26 (113.10) AU/mL, respectively. The level of serum IgG antibody in the pneumonia group was lower than that of the non-pneumonia group at T1 and T4 time points, and the differences were statistically significant (P<0.05) , the positive rate of serum IgG antibody in the pneumonia group was lower than that of the non-pneumonia group at the T1 time point, and the difference was statistically significant (P<0.05) . The serum IgM antibody levels at 5 time points were 0.41 (0.81) AU/mL, 0.95 (1.62) AU/mL, 1.09 (2.42) AU/mL, 0.74 (3) AU/mL, and 0.81 (3.10) AU/mL respectively, and there was no significant difference between the two groups. Conclusion　The clinical symptoms of patients infected with the SARS-CoV-2 B.1.1.529 variant strain are mild. Serum IgG antibodies increased after infection, but there are some differences between the pneumonia group and the non-pneumonia group, whether serum IgG has a protective effect needs further research; the serum IgM antibodies do not increase highly after infection, there are some differences between individuals.

The TCM (traditional Chinese medicine) transdermal plaster (also known as "cataplasma") are flexible adhesive patches used for treatment of pain, resulted from arthritis, sprain and bruise, tendovaginitis, lumbar spine protrude, neuralgia, hyperosteogeny ache, abdominal discomfort and metastatic cancer, etc. Since the 1980's, investigators in China have used this modern patch delivery system for herbal drugs and obtained satisfactory results especially from the treatment of various types of pain associated with bone diseases, abdominal discomfort, and tumors, etc. The production of TCM cataplasma was successfully scaled up in early 90's and the commercial product line for an antirheumatic agent was first established in Shanghai by Leiyunshang Group. Thus far, a number of products in the form of TCM cataplasma became commercially available in the market, and clinical investigations with these products indicated that topically applicable herbal preparations, especially in the form of cataplasma, are preferred formulations with respect to the treatment comfort of the patient. Compared to the traditional preparations which utilize rubber and rosin as adhesives, cataplasma is advantageous in that the lipophilic and hydrophilic ingredients of the herbal extracts are solubilized and then "gellified" with the organic polymers, and that the drug matrix containing up to 40%-70% of water serves as a "drug reservoir" that will sustain the quick and continuous release of herbal ingredients over several days across the skin. While there are conventional remedies for palliation of pain and discomfort associated with bone diseases or cancers, administration of oral medicinal herbs combined with topical agents such as TCM cataplasma may significantly alleviate the symptoms and improve their quality of life. This article provides a review on three aspects, which include the process development, characteristics and developmental status of TCM cataplasma, and future development of such a technology.
Administration, Cutaneous ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; therapeutic use ; Bone Neoplasms ; complications ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; Pain ; drug therapy ; etiology ; Phytotherapy ; Skin Absorption ; Technology, Pharmaceutical ; methods

To explore the mechanism of the absorption enhancement of Angelica dahurica extract (Ade), the absorption mechanism of baicalin in the Scutcllaria water extraction as well as the effect of Angelica dahurica extract on absorption of baicalin were investigated. In order to determine the main absorption site, everted intestinal sac model was used to study the effect of Angelica dahurica extract on the absorption of baicalin at duodenum, jejunum, ileum and colon. In situ single pass intestinal perfusion model was performed to study the absorption of various concentrations of baicalin and the effect of Angelica dahurica extract on the absorption of baicalin at the main absorption site. To authenticate the consequence of perfusion by getting the blood from the hepatic portal vein and determine the concentration of the baicalin in the blood. The result showed that baicalin could be absorbed at all of the four intestinal segments with increasing absorption amount per unit as follows: ileum > colon > jejunum > duodenum. The absorption ofbaicalin in the duodenum significantly increased with Angelica dahurica extract, thus, duodenum was chosen to be the studying site. Apparent permeability values (Papp) and absorption rate constant (Ka) of baicalin in the duodenum increased gradually with higher concentrations. When the concentration of baicalin rises to a certain degree, the absorption increase had a saturable process, the absorption of baicalin may be an active transportation. Baicalin may be not a substrate of P-gp as verapamil which had not significantly affected the Papp and Ka of baicalin. The absorption of baicalin in the duodenum significantly increased (P < 0.01) in the two models with Angelica dahurica extract and the concentration of baicalin in the blood from the hepatic portal vein showed that the Angelica dahurica extract can increase the absorption of baicalin.
Angelica ; chemistry ; Animals ; Drug Synergism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Duodenum ; metabolism ; Flavonoids ; isolation & purification ; pharmacokinetics ; Herb-Drug Interactions ; Intestinal Absorption ; drug effects ; Intestines ; metabolism ; Male ; Perfusion ; Permeability ; Plants, Medicinal ; chemistry ; Portal Vein ; metabolism ; Rats ; Rats, Sprague-Dawley ; Scutellaria ; chemistry ; Verapamil ; pharmacology

Objective:To evaluate the efficacy of 3D printed individualised prosthesis in treating bone and joint defects in upper limbs remained after earlier microsurgical repairs.Methods:From June 2019 to September 2021, 12 patients were treated in the Institute of Orthopaedic Trauma of PLA, the 80th Group Army Hospital for bone and joint defects in upper limb that had been remained after earlier repairs with soft tissue flaps. The defects were: 1 in completely severed wrist, 2 defects of digit metacarpal bone, 4 defects of interphalangeal joint, 4 defects of bones in radiocarpal joint and 1 defect of lunate bone. The area of soft tissue defect ranged from 1.5 cm×3.0 cm to 12.0 cm×18.0 cm, and the length of bone defects ranged from 2.5 to 8.5(average 3.64) cm. For incompletely severed and completely severed limbs, replantation of severed limbs (digits) were performed in the primary surgery and the repair of soft tissue defects were performed in the second stage surgery. The remaining defects of bone and joint were reconstructed by 3D printed individualised prostheses in the third stage surgery. Finger soft tissue defects were covered with a local flap in the primary surgery, and bone and joint defects were reconstructed with a 3D printed prosthesis in the second surgery. Finger soft tissue defects were covered with a local flap in the first phase, and bone and joint defects were reconstructed with a 3D printed prosthesis in the second phase. After the surgery, the bone integration between the broken end of the bone joint defect and the prosthesis was determined based on the X-ray results and the Paley fracture healing score standard. Simultaneously measured the Total Active Motion(TAM) of the forearm and hand joints. At 1, 2, 3, 6, 9 and 12 months after hospital discharge. Follow-up X-ray examinations were taken followed by examinations on the recovery of soft tissues and bones. The upper limb function was graded according to the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association.Results:Postoperative follow-up at outpatient clinic lasted for 6 to 26 months, with an average of 11.5 months. All flaps were free from necrosis and infection, also there was no infection in bones and joints. According to the Paley fracture healing scale, 10 patients were in excellent and 2 in good. In addition, according to the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association, 5 patients achieved upper limb function in excellent, 5 in good and 2 in fair. The ranges of motion of the affected wrists were 30°-42°(average 37.3°) for the implanted prostheses of distal end of radius and the radial shaft. Wrist flexion 40° to 55°(average 43.5°). The range of motion of finger and wrist was 60° to 70°(average 65.7°) with a metacarpal and phalangeal bone prosthesis.Conclusion:3D printed individually customised prostheses are safe, accurate and effective in repair of the remained bone and joint defects in upper limbs after primary and early stages of microsurgical flap repairs. It can effectively restore anatomical structures of bone and joint in upper limbs.

OBJECTIVETo prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy.

METHODSCT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50,000 and above 300,000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70,000, 90,000, 95,000, 110,000 and 170,000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL.

RESULTSMajor of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK(P<0.01).

CONCLUSIONSThe rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42 centi-degree heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.

Animals ; Cancer Vaccines ; immunology ; Cells, Cultured ; Heat Stress Disorders ; metabolism ; Heat-Shock Proteins ; metabolism ; Mice ; Mice, Inbred BALB C ; Molecular Chaperones ; immunology ; Trichosanthin ; pharmacology

OBJECTIVETo explore the application of biosensor technology in the determination of endotoxin-neutralizing materials.

METHODSAfter mixing polymyxin B (PMB) with endotoxin in certain concentration, the neutralizing ratio of PMB to endotoxin was assessed by biosensor technique and limulus amebocyte lysate test respectively. The results from the two methods were compared.

RESULTSThe neutralizing ratio of PMB to endotoxin as assessed by biosensor technology was 0.35 microg to 1 ng, while that by dynamic turbidimetric and chromogenic limulus amebocyte lysate (LAL) technique was 0.5 mg to 1 ng and 1 mg to 1 ng, respectively. The results obtained by biotechnology were similar to that by biosensor technique.

CONCLUSIONBiosensor technology was an accurate, convenient and rapid method for the determination of potency of endotoxin-neutralizing materials.

Bacterial Proteins ; analysis ; Biosensing Techniques ; methods ; Endotoxins ; analysis ; Lipid A ; analysis ; Polymyxin B ; analysis ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVETo investigate the possibility of crosstalk between phosphatidylinositol 3-kinase (PI3-K)/Akt pathway and p38 mitogen-activated protein kinase (p38MAPK) pathway in cardiomyocyte with challenge of burn serum, and to explore their influence on cardiomyocyte injury after burn.

METHODSThe model of murine cardiomyocyte with stimulation of burn serum was established. (1) The level of Akt and p38 phosphorylation in cardiomyocyte were examined with stimulation of 10% burn serum before stimulation and 1, 3, 6, 12, 24 hour after stimulation. (2) The levels of Akt and p38 phosphorylation in cardiomyocyte were determined with stimulation of burn serum (at concentration of 5%, 10%, 20%) or 10% burn serum plus insulin (at concentration of 1 x 10(-8), 1 x 10(-7), 1 x 10(-6)mol/L). The content of creatine kinase (CK) in supernate was also detected. (3) Addition to the inhibitor of p38 MAPK pathway (SB203580) and PI3K/Akt pathway (LY294002), the level of p38MAPK, PI3K/Akt and the content of CK in supernate were determined.

RESULTS(1) The level of p-p38 in cardiomyocyte was 4.0 +/- 0.8, 3.6 +/- 0.8, 5.1 +/- 1.6, 2.4 +/- 0.5, 3.0 +/- 0.6 at 1, 3, 6, 12, 24 hour (s) after stimulation of burn serum, which was obviously higher than that immediate after stimulation (1.0, P < 0.01). The level of p-Akt was 0.15 +/- 0.07, 0.64 +/- 0.10, 0.26 +/- 0.08, 0.38 +/- 0.11, 0.59 +/- 0.13, which was obviously lower than that before stimulation (1.00, P < 0.01). (2) With stimulation of different concentration of burn serum or burn serum plus insulin, the level of p-Akt and p-p38 changed in the opposite directions comparatively. The content of CK increased along with increase of burn serum concentration, but decreased obviously with treatment of insulin (P < 0.05 or 0.01). (3) Low level of p38 induced by burn serum was increased after treatment of LY294002, which neutralized the protection of insulin (P < 0.01). Low level of p-Akt induced by burn serum increased after treatment of SB203580 (P < 0.01), which inhibited the release of CK induced by burn serum.

CONCLUSIONThere is being crosstalk between PI3K/Akt pathway and p38 MAPK pathway in cardiomyocytes with challenge of burn serum, which may regulate cardiomyocytes.

Animals ; Burns ; blood ; Male ; Myocytes, Cardiac ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serum ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the protective effect of insulin on oxygen-radical induced hepatic injury in severely scalded rats in early stage of severe scald.

METHODSEighty-four male Sprague-Dawley rats were randomly divided into three groups: i. e, normal group, saline group, and insulin group, with 28 rat in each group. The rats in the latter two groups were subjected to 30% TBSA full-thickness scald on the back, and received intra-peritoneal injection of 40ml/kg isotonic saline, and subcutaneous injection of 3 IU/kg insulin, respectively. The total anti-oxygen capability (T-AOC), the expression of superoxide dismutase (SOD), reactive oxygen species (ROS) and intercellular adhesion molecule (ICAM-1) in hepatic tissue, and serum alanine transaminase (ALT) were determined in each group at 6, 12, 24, 48 post-scald hours (PSH) with corresponding methods.

RESULTSThe hepatic T-AOC and SOD content were obviously decreased, while the ROS content were markedly increased at 6 PSH in saline group compared with that in normal group (P < 0.05 or P < 0.01). The expression of ICAM-1 and serum content of ALT were significantly higher than that in normal group at 12 PSH and 48 PSH (P < 0.01). At 24 PSH, the hepatic T-AOC (386 +/- 75) U/g and SOD content (210 +/- 39 ) U/g were obviously higher in insulin group than those in saline group [(124 +/- 18), (111 +/- 9) U/g, respectively, P < 0.01), but the ROS content (154 +/- 29 ) U/g was much lower than that in saline group [(351 +/- 41) U/g, respectively, P < 0.01]. At 48 PSH, the serum content of ALT and hepatic expression of ICAM-1 in insulin group exhibited obvious difference when compared with those in saline group (P < 0.01). Meanwhile, Pathological examination showed that hepatic injury was alleviated by insulin administration after scald.

CONCLUSIONInsulin administration early after severe scald exhibits protective effect on liver function by improving anti-oxygen radical ability of rat liver.

Alanine Transaminase ; blood ; Animals ; Burns ; metabolism ; pathology ; Insulin ; pharmacology ; Liver ; drug effects ; metabolism ; pathology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism