Objective To verify adhesion and growth ability of canine esophageal epithelial cells (EECs) on the poly (lactic-co-glycolic acid) (PLGA), a three-dimensional biodegradable polymer scaffold, and to reconstruct the canine esophagus by the tissue engineering. Methods Free canine EECs isolated from adult dogs by esophagoscopy were seeded onto the PLGA scaffolds precoated with collagen type Ⅳ after the first passage by the in vitro culture. Then, the composites of the cell-scaffold were respectively cultured invitro and in the abdominal cavity of the dog in vivo. After different periods, the cell-seeded scaffolds were assessed by histological HE staining, scanning electron microscopy, and immunohistochemical analysis. Results The cells displayed a cobblestone-shaped morphology that was characteristic of the epithelial cells and were stained to be positive for cytokeratin, which indicated that the cells were EECs. The canine EECs were well distributed and adhered to the PLGA scaffolds, and maintained their characteristics throughout the culture period. After the culture in vivo for 4 weeks, the cell-seeded scaffolds looked like tissues. Conclusion PLGA scaffolds precoated with collagen type Ⅳ can be suitable for adhesion and proliferation of EECs, and can be used as a suitable tissue engineering carrier of an artificial esophagus.
Objective To investigate the controlled release effect and the anti-cancer cell ability of a 5-FU loaded poly-L-lactic acid?（PLLA） nanofibers membrane blending with keratin. Methods Making PLLA and keratin mix together and crosslinking to generate blending solution. Then the anti-cancer drug 5-FU was added into the solution to fabricate nanofibers membrane by high voltage electrospinning method. The micro morphology was observed by scanning electron microscope （SEM）. The controlled release effect of 5-FU from the nanofibers membrane was measured by high performance liquid chromatography （HPLC）. The cytotoxicity of 5-FU/PLLA keratin nanofibers membrane was evaluated by using 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide （MTT） assay on HCT116 cell lines. At the meantime, cell growth morphology of HCT116 in experiment group were observed by microscope and transmission electron microscope. Results 5-FU could be dispersed homogeneous in the PLLA/keratin nanofibers membrane through SEM. HPLC suggested that 5-FU could be diffused out from the fibers slowly and uniformly, which corresponded the zero order kinetics basically. After different treatment, the longer time the 5-FU/PLLA keratin nanofibers （experiment group） immerse in the medium, the much more swelling, apoptosis, and necrocytosis of the cells were observed. The cell viability for experiment group was （47.5±2.8）% by MTT, while the PLLA keratin nanofibers without 5-FU had no significant impact on cell viability （93.9±2.8）%, which was statistic significance （P＜0.01）. Conclusion 5-FU/PLLA keratin nanofibers membrane owns good controlled release effect and satisfies cell inhibitory effect against HCT116 cells in vitro，which suggested that it has a promising prospect for clinical therapy.
Objective To explore the clinical characteristics of patients with lymphoma firstly manifested as symptoms in respiratory system. Methods The clinical data of 9 patients with lymphoma were analyzed retrospectively and discussed with literature review. Results There were 7 males and 2 females with an average age of 48.2 years and a median disease course of 20 days. All patients were diagnosed by pathology while specimens were obtained by cervical lymph node biopsies in 2 cases, by CT-guided percutaneous lung biopsies in 2 cases, by bronchoscope mucosal biopsies in 2 cases, by transbronchial needle aspiration biopsies in 1 case, by thoracoscope lung biopsies in 1 case, and by ascites cell block inspection in 1 case. The main symptoms were cough (7/9), expectoration (3/9), fever (3/9) and wheeze (2/9). Chest CT showed tumors (3/9, multiple in 1 case), enlarged mediastinal lymph nodes (6/9), enlarged hilar lymph nodes (3/9), pulmonary consolidation (3/9), pleural effusion (6/9, bilateral in 3 cases), pleural thickening (2/9), pulmonary atelectasis (2/9), patchy shadow (7/9), pericardial effusion (1/9). Laboratory examination demonstrated elevated cancer antigen 125 (CA125) in 7 cases while elevated lactic acid dehydrogenase (LDH) in 4 cases. One patient died during hospitalization in the respiratory department, 1 patient auto-discharged without further treatment, 1 patient died in follow-up period. Five patients remain alive up to now. Conclusions The symptoms of patients with lymphoma are atypical while the chest radiological findings are varied. CA125 and LDH play important role in evaluating disease and predicting prognosis in patients with lymphoma.
Objective To evaluate the internal fixation effect, degradation, and biocompatibility of polylactic-co-glycolic acid/hydroxyapatite (PLGA/HA) absorbable cannulated screws in treatment of lateral femoral condyle fracture of canine so as to provide the theory basis for their further improvement and clinical application. Methods Sixteen adult male Beagles (weighing, 9-12 kg) were selected to prepare the models of bilateral lateral femoral condyle fracture; left fracture was fixed with PLGA/HA absorbable cannulated screws as experimental group and right fracture with metal screws as control group. At 2, 4, 8, and 12 weeks after operation, general observation was done and X-ray films were taken for observing fracture healing; bone mineral density was measured; the histological examination was performed; and the degradation property of absorbable cannulated screws was detected. Results All animals survived to the end of the experiment. General observations showed that no fracture displacement occurred and fracture healed at 12 weeks in 2 groups; no breakage, displacement, or loosening of screws was observed in experimental group. X-ray films results showed that the absorbable cannulated screws could not be found out by X-ray in experimental group, but metal screws could be found out in control group; fracture healed with time in 2 groups. The bone mineral density reached the peak at 8 weeks in 2 groups, and no significant difference was found between 2 groups and among different time points in the same group (P gt; 0.05). Histological examination showed that 2 groups had similar fracture healing process at different time points; no obvious inflammatory reaction was found around absorbable cannulated screws in experimental group. The degradation results of absorbable cannulated screws showed that the intrinsic viscosity and molecular weight distribution obviously decreased at 2 weeks; the number average molecular weight and the weight average molecular weight markedly decreased at 4 weeks; and the maximum shear force did not decrease obviously at 8 weeks, and then decreased significantly. Significant differences were found in all indexes among different time points in the same group (P lt; 0.05). Conclusion PLGA/HA absorbable cannulated screws and metal screws show similar fracture healing process for fixing lateral femoral condyle fracture of canine, and the absorbable canulated screws have good biocompatibility. The maximum shear force of PLGA/HA absorbable cannulated screw has no obvious decrease during 8 weeks after operation, so it can ensure full healing of fracture.
ObjectiveTo investigated the levels of aldolase A (ALDOA) in pleural effusion in patients with different pathological types of lung cancer and patients with tuberculous pleurisy,and the correlation between ALDOA and carcinoembryonic antigen (CEA),lactate dehydrogenase(LDH). Methods80 cases of pleural effusion samples were collected,of which 65 cases of lung cancer (malignant group) and 15 cases of tuberculous pleurisy (TB group). All the patients were not treated with anti-inflammatory or steroid therapy. ALDOA concentrations in pleural effusion were detected by ELISA and the contents of CEA and LDH in pleural fluid were detected by chemiluminescence assay. ResultsThe levels of ALDOA,CEA and LDH in the malignant group were 46.75±21.39 ng/mL,82.24±56.63 ng/mL,755.76±382.54 U/L respectively,and were 23.92±17.21 ng/mL,2.55±1.67 ng/mL,and 388.37±163.87 U/L in the TB group respectively. The levels of ALDOA,CEA and LDH in the malignant group were significantly higher than those in the TB group (P<0.01). The concentrations of ALDOA in malignant pleural effusion from different pathological types of lung cancer were 71.65±32.09 ng/mL(adenocarcinoma),22.43±18.23 ng/mL(small cell lung cancer),and 19.16±13.85 ng/mL(squamous cell carcinoma),respectively. The concentration of ALDOA in malignant pleural effusion from the adenocarcinoma patients was significantly higher than that in the other two types (P<0.05). The concentration of CEA was 112.40±62.71 ng/mL(adenocarcinoma),62.45±54.78 ng/mL(small cell lung cancer),and 71.87±52.4 ng/mL(squamous cell carcinoma),respectively. It was significantly higher in adenocarcinoma than that in other two types (P<0.05). The levels of LDH were 661.81±328.93 U/L(adenocarcinoma),737.62±315.41 U/L(small cell lung cancer),767.85±503.28 U/L(squamous cell carcinoma),respectively. There was no significant difference in three types(P>0.05). The concentrations of ALDOA in pleural effusion from the patients with lung cancer or tuberculous pleurisy were positively correlated with the concentrations of CEA and LDH (P<0.01 or 0.05). ConclusionThe levels of ALDOA,CEA and LDH in malignant pleural effusion from lung cancer patients were significantly higher than those in pleural effusion from patients with tuberculous pleurisy. The ALDOA and CEA levels in malignant pleural effusion from lung adenocarcinoma patients were significantly higher than those in small cell lung cancer and squamous cell carcinoma patients. There were highly positive correlation between ALDOA,CEA and LDH levels.
ObjectiveTo review the research progress and challenges of poly (L-lactic acid) (PLLA) membrane in preventing tendon adhesion. MethodsThe relevant literature at home and abroad in recent years was extensively searched, covering the mechanism of tendon adhesion formation, the adaptation challenge and balancing strategy of PLLA, the physicochemical modification of PLLA anti-adhesion membrane and its application in tendon anti-adhesion. In this paper, the research progress and modification strategies of PLLA membranes were systematically reviewed from the three dimensions of tissue adaptation, mechanical adaptation, and degradation adaptation. ResultsThe three-dimensional adaptation of PLLA membrane is optimized by combining materials (such as hydroxyapatite, polycaprolactone), structural design (multilayer/gradient membrane), and drug loading (anti-inflammatory drug). The balance between anti-adhesion and pro-healing is achieved, the mechanical adaptation significantly improve, and degradation is achieved (targeting the degradation cycle to 2-4 weeks to cover the tendon repair period). ConclusionIn the future, it is necessary to identify the optimal balance point of three-dimensional fitness, unify the evaluation criteria and solve the degradation side effects through the co-design of physicochemical modification and drug loading system to break through the bottleneck of clinical translation.
ObjectiveTo investigate the clinical values of serum histidine decarboxylase (HDC), D-lactate, and alpha-glutathione S-transferase (α-GST) for diagnosing intestinal mucosal injury of patients with intestinal obstruction. MethodsThe expression levels of serum HDC, D-lactate, and α-GST in 28 patients with strangulated intestinal obstruction, 19 patients with simple intestinal obstruction, 17 patients with acute simple appendicitis, and 20 healthy volunteers were determined by enzyme linked immunosorbent assay (ELISA) before the treatment, and then the area under receiver operating characteristic curve (AUC) of these diagnostic indices were compared. In addition, the occurrence rates of systemic inflammatory response syndrome (SIRS) and infectious complications (abdominal cavity infection and pulmonary infection) were closely observed. The relevances of SIRS and infectious complications and the expression levels of these three diagnostic indices were analyzed. ResultsThe expression levels of serum HDC, D-lactate, and α-GST of the patients with strangulated intestinal obstruction were the highest among all the patients (Plt;0.01), and the expression levels of these three indices in the patients with simple intestinal obstruction were higher than those of the patients with acute simple appendicitis (Plt;0.05). The AUC of HDC (0.913) was larger than that of D-lactate (0.872) and α-GST (0.836) (P=0.000, P=0.000, respectively). When the cut off value of HDC was 31.00 μg/L, the sensitivity, specificity, false negative rate, and false positive rate of HDC were 74.5%, 94.6%, 25.5%, and 5.4%, respectively, which were all better than those of D-lactate and αGST. The occurrence rates of SIRS and abdominal cavity infection of the patients with strangulated intestinal obstruction were significantly higher than those of patients with simple intestinal obstruction (P=0.046) and acute simple appendicitis (P=0.027); while there was not significantly different of pulmonary infection among all the patients (P=0.728). The expression level of serum HDC in patients with strangulated intestinal obstruction suffered from SIRS (P=0.000) or abdominal cavity infection (P=0.002) was significantly higher than that of not-suffered from SIRS or uninfected patients. Meanwhile, the expression levels of serum D-lactate and α-GST in the patients with strangulated intestinal obstruction suffered from SIRS were higher than those of notsuffered from SIRS patients (P=0.032, P=0.021, respectively). The expression levels of HDC, D-lactate, and α-GST were significantly correlated with SIRS and abdominal cavity infection (Plt;0.05), among which the level of HDC and the incidence of SIRS had the highest correlation (r=0.608, P=0.001). ConclusionHDC may be a more effective index for diagnosing intestinal mucosal injury of patients with intestinal obstruction.
ObjectiveTo observe the effect of vascular endothelial growth factor/polylactide-polyethyleneglycol-polylactic acid copolymer/basic fibroblast growth factor (VEGF/PELA/bFGF) mixed microcapsules in promoting the angiogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. MethodsThe BMSCs were isolated by the method of whole bone marrow adherent, and sub-cultured. The passage 3 BMSCs were identified by Wright-Giemsa staining and flow cytometry, and used for subsequent experiments. VEGF/PELA/bFGF (group A), PELA/bFGF (group B), VEGF/PELA (group C), and PELA (group D) microcapsules were prepared. The biodegradable ability and cytotoxicity of PELA microcapsule were determined, and the slow-released ability of VEGF/PELA/bFGF mixed microcapsules was measured. The passage 3 BMSCs were co-cultured with the extracts of groups A, B, C, and D, separately. At 1, 3, 7, 14, and 20 days after being cultured, the morphological changes of induced BMSCs were recorded. At 21 days, the induced BMSCs were tested for DiI-labeled acetylated low density lipoprotein (Dil-ac-LDL) and FITC-labeled ulex europaeus agglutinin I (FITC-UEA-I) uptake ability. The tube-forming ability of the induced cells on Matrigel was also verified. The differences of the vascularize indexes in nodes, master junctions, master segments, and tot.master segments length in 4 groups were summarized and analyzed. ResultsThe isolated and cultured cells were identified as BMSCs. The degradation time of PELA was more than 20 days. There was no significant effect on cell viability under co-culture conditions. At 20 days, the cumulative release of VEGF in the mixed microcapsules exceeded 95%, and the quantity of bFGF exceeded 80%. The morphology of cells in groups A, B, and C were changed. The cells in groups A and B showed the typical change of cobble-stone morphology. The numbers of double fluorescent labeled cells observed by fluorescence microscope were the most in group A, and decreases from group B and group C, with the lowest in group D. The cells in groups A and B formed a grid-like structure on Matrigel. Quantitative analysis showed that the differences in the number of nodes, master junctions, master segments, and tot.master segments length between groups A, B and groups C, D were significant (P<0.05). The number of nodes and the tot.master segments length of group A were more than those of group B (P<0.05). There was no significant differences in the number of master junctions and master segments between group A and group B (P>0.05). ConclusionVEGF/PELA/bFGF mixed microcapsules have significantly ability to promote the angiogenic differentiation of rat BMSCs in vitro.
Objective To observe the release pattern of the microcysts and the effect of ectopic osteogenesis of combined micromorselized bone by optimized preparation of microcysts. Methods Optimized poly-DLlactide-co-glycolide (PLGA) microcysts manufacturing method was performed with the orthogonal design, and the accumulated release amount of microcysts was calculated at 2 h, 4 h, 8 h, 12 h, 24 h, 36 h, 48 h, 60 h, 72 h, 84 h, 96 h, 120 h, 144 h, 168 h, 192 h, 216 h, 240 h and 264 h. Twentyfour Wistar rats were divided into 4 groups (n=6) and 1 cm length incision was cut in their bilateral thighs skin, forming 48 gluteus maximus muscle sackmodels. In group A,collagen was implanted to bilateral muscle sacks respectively. In group B, collagen and autologous morselized bone were implanted to bilateral muscle sacks. Ingroup C, collagen and rhBMP-2/PLGA delayed release microcysts were implanted to bilateralmuscle sacks respectively. In group D, collagen and morselized bone/rhBMP-2/PLGA delayed release microcysts were implanted to bilateral muscle sacks. Gross and histologic observations were made at 3, 4 and 5 weeks postoperatively.Results Every optimized variance had an effect on particle diameter of microcyst and its encapsulating rate. The microcyst’s surface was smooth and had a fine spheroplast, which released slowly within 11 days in vitro. In thethird week postoperatively, the graft in group A could not be touched, while the graft in all other 3 groups was still found. After 3 weeks, collagen was absorbed completely in group A, the residual collagen could be seen in groups B, C andD. After 4 weeks, collagen could be seen in group A; micromorselized bone continued to be absorbed and became smaller in group B; microsphere became smaller, osteoblasts increased in group C; micromorselized bone and microsphere continuedto be absorbed, oteoblasts and chondroblasts increased. After 5 weeks, implantsbecame small, microsphere was absorbed, osteoblasts and chondroblasts became more in groups B, C and D. Microcysts presented with white granuloshape and were packaged in tissue pieces. Histologic observation showed that the PLGA microcysts in 3 weeks and 4 weeks could be absorbed gradually as the time in vivo, if combining with morselzed bone they could produce abundant induced osteoblasts and chondroblasts. Conclusion Optimizing the preparation technology of microcysts has delayed their release during a long period in vitro. Autologous micromorselized bone can be ectopicly induced to produce large amount of osteoblasts in gluteus maximus muscle sack, where PLGA microcysts can combine organically and bring about the bone formation with less amount of growth factors.
Objective To observe the therapeutic effect of poly tetrahydrofurfuryl co-lactic acid(copolymer C4) as the biodegradable vitreous substitutes on rhegmatogenous retinal detachment.Methods Vitreoretinal surgery with copolymer C4 tamponades was performed on 32 pigmented rabbits (64eyes) with rhegmatogenous retinal detachment. The rate of reattached retina and the post operative cornplications were observed.Results Three months after the operation, reattached retina was found in 96. 4%, glaucoma in 5.5%, cataract in 10.9%, and copolymer emulsion in 10.2% of all the eyes.Conclusion copolymer C4 may withstand the retinal tear effectively for 3 months, and can be a valuable vitreous substitutes. (Chin J Ocul Fundus Dis，2004，20：27-28)