Objective To investigate the preventive effect of carbachol on the formation of postoperative intra-abdominal adhesion. Methods Forty-four Wistar rats were randomly divided into sham operation group (SO group, n=12), operation group (n=16) and carbachol treated group (carbachol group, n=16, carbachol 50 μg/kg). Animal model of abdominal adhesion was established by rubbing the procussus vermiformis of cecum with dry sterile gauze, and by clamping and scuffing abdominal wall. Half of rats were separately killed on day 7 and day 14 after surgery, respectively. The degree of adhesion was evaluated according to Phillips 5-scale grade and the feature of this model. The histopathological changes of adhesive tissues were observed and the content of collagen type Ⅰ in the tissues was detected by immunohistochemistry. Results The scores of intra-abdominal adhesion were significantly lower in the carbachol group than those in operation group both on 7 d and 14 d (P＜0.01). Mild inflammatory changes and less fibrous proliferation were observed in carbachol group microscopically. The contents of collagen type Ⅰ detected by immunohistochemistry were significantly lower in the carbachol group than those in operation group both on 7 d and 14 d (P＜0.01). There was no significant difference of the score of abdominal adhesion and content of collagen type Ⅰ in the same group between 7 d and 14 d (Pgt;0.05). Conclusion Carbachol may take a significant role in the prevention of postoperative abdominal adhesion in rat.
OBJECTIVE The effect of platelet-derived wound healing factor (PDWHF) on wound healing in diabetic rats was studied. METHODS Forty-four male SD rats were randomly divided into 2 groups. Thirty-two rats of experimental group accepted intraperitoneal injection of alloxan (1.5 mg/10 g body weight). Within one or two days after injection, while the blood sugar of the rats was higher than 180 mg/dl, the animal model of diabetic rat should have been established. Then a dorsal incision was given to every rat. After the addition of PDWHF (the experimental group) or bovine albumin (the control group), the incision was sutured up. Seven, ten and fourteen days after operation, the breaking strength of the wound was measured. On another hand, specimen from the wound was taken for the culture of fibroblasts. When the cultured fibroblasts have been incubated with 10% PDWHF for 4, 8 and 12 hours, the procollagen I (alpha 1) mRNA levels were examined respectively, and compared with those of control. RESULTS Significant difference in wound breaking strength had been observed between PDWHF-treated incisions and the control on 7, 10 and 14 days after wounding (P lt; 0.01). Experiment in vitro demonstrated that the procollagen I (alpha 1) mRNA levels in wound fibroblasts incubated with 10% PDWHF for 4, 8 and 12 hours were 0.9, 3.7 and 2.2 folds higher than those in fibroblasts in control. CONCLUSION It was suggested that direct stimulation of procollagen I (alpha 1) gene expression was one of the ways that PDWHF played its role in accelerating wound healing.
Objective To investigate the effect of collagen type I concentration on the physical and chemical properties of the collagen hydrogel, and to analyze the effect of different concentrations of collagen type I hydrogel on the phenotype and gene expression of the chondrocytes in vitro. Methods Three kinds of collagen hydrogels with concentrations of 12, 8, and 6 mg/ mL (C12, C8, and C6) were prepared, respectively. The micro-structure, compressive modulus, and swelling ratio of the hydrogels were measured and analyzed. The chondrocytes at 2nd passage were cocultured with three kinds of collagen hydrogels in vitro, respectively. After 1-day culture, the samples were stained with fluorescein diacetate (FDA) / propidium iodide (PI) and the cell activity was observed under confocal laser microscope. After 14-day culture, HE staining and toluidine blue staining were carried out to observe the histological morphology, and mRNA expressions of chondrocytes related genes (collagen type II, Aggrecan, collagen type I, collagen type X, Sox9) were determined by real-time fluorescent quantitative PCR. Results With the increase of collagen type I concentration from 6 to 12 mg/mL, the physical and chemical properties of the collagen hydrogels changed significantly: the fiber network became dense; the swelling ratios of C6, C8, and C12 were 0.260 ± 0.055, 0.358 ± 0.072, and 0.539 ± 0.033 at 192 hours, respectively, showing significant differences among 3 groups (P lt; 0.05); and the compression modulus were (4.86 ± 0.96), (7.09 ± 2.33), and (11.08 ± 3.18) kPa, respectively, showing significant differences among 3 groups (P lt; 0.05). After stained with FDA/PI, most cells were stained green, and few were stained red. The histological observation results showed that the chondrocytes in C12 hydrogels aggregated obviously with b heterochromia, chondrocytes in C8 hydrogels aggregated partly with obvious heterochromia, and chondrcytes in C6 hydrogels uniformly distributed with weak heterochromia. Real-time fluorescent quantitative PCR results showed that the mRNA expressions of collagen type II and Aggrecan were at the same level in C12, C8, and C6; the expressions of collagen type I, Sox9, and collagen type X were up-regulated with the increase of collagen type I hydrogels concentration, and the expressions were the highest at 12 mg/mL and were the lowest at 6 mg/mL, showing significant differences among 3 groups (P lt; 0.05). Conclusion Increasing the concentration of collagen hydrogels leads to better mechanical properties and higher shrink-resistance, but it may induce the up-regulation of cartilage fibrosis and hypertrophy related gene expression.
Objective To investigate the effect of vanadate on prol iferation and collagen type I production of rat medial collateral l igament (MCL)fibroblasts. Methods A total of 12 adult male SD rats were included, weighing 350-375 g. MCL was cut into small pieces (1 mm × 1 mm × 1 mm) in aseptic conditions, and then placed and cultured in culture chambers. Fibroblasts were passaged with 0.25% trypsin. The vanadate (0, 1.0, 2.5, 5.0 ng/mL) was added in the 3rd passage MCL fibroblasts, respectively, and the samples were divided into 4 groups (0, 1.0, 2.5, 5.0 ng/mL groups). MTT was used to measure the cell prol iferation. The production of collagen type I was measured by RT-PCR and ELISA. Twelve samples in each group were measured. Results In fibroblast prol iferation, the absorbency values of the 1.0, 2.5, 5.0 ng/mL groups were significantly different from that of the 0 ng/mL group (P lt; 0.05). The absorbency values of the 0, 1.0, 2.5, and 5.0 ng/mL groups were 0.213 ± 0.016, 0.327 ± 0.023, 0.449 ± 0.137, and 0.561 ± 0.028, respectively. In collagen secretion, vanadate in 1.0, 2.5, 5.0 ng/mL groups could significantly induce the production of collagen type I (P lt; 0.05) ina dose-dependent manner. The expressions of collagen type I of 0-5 ng/mL groups were 0.47 ± 0.02, 0.51 ± 0.03, 0.60 ± 0.01, and 0.72 ± 0.02, respectively. There was significant difference between the 1.0, 2.5, 5.0 ng/mL groups and 0 ng mL group (P lt; 0.05). RT-PCR displayed a dramatic increase of band density. The ratio of band density in the 0-5 ng mLgroups was 1.37 ± 0.76, 1.97 ± 0.53, 2.41 ± 0.94, and 2.73 ± 0.82, respectively. The gene expression of collagen type I in the 1.0, 2.5 and 5.0 ng/mL groups was higher than that in the 0 ng/mL group, and there was significant difference (P lt;0.05). There were statistical significant differences among 1.0, 2.5 and 5.0 ng/mL groups in each index mentioned above.Conclusion Vanadate can effectively induce the prol iferation of fibroblasts and the production of collagen type I in vitro, which may provide a new approach to the treatment of MCL injury.
Objective To explore the feasibility of combining high level Le Fort Ⅰ osteotomy with bone grafting in the same operation for correction of secondary midface deformities in cleft patients. Methods From January 2002 to January 2005, 10 patients suffering from secondary midface deformities were treated. There were 4 males and 6 females, aged from 16 to 32 years. The unilateral cleft was involved in 8 patients and the bilateral cleft in2 patients. All patients received combining high level Le Fort Ⅰ osteotomy with bone grafting in the same operation. The horizontal corticotomy of high level Le Fort Ⅰosteotomy on anterior wall of maxillary bone is higher than that of traditionalLe Fort Ⅰ osteotomy, it is only 5 mm close to infraorbital foramen. Results All 10 patients were satisfied with their appearances after operation. Dental articulation was improved greatly in 9 patients. With an X-ray re-examination, maxillary was returned to normal position in all patients. After a follow up from 6months to 2 years, dental arch had good appearance. The X-ray films showed no obvious bone absorption. The density of grafting bone was approximation to the normal bone.Conclusion High level Le Fort Ⅰ osteotomy can make notonly maxillary advance, but also regions of lateral and floor of nose and partial infraorbital region advance. Combining with bone grafting in the same operation can decrease the frequency of operation, save the treatment expense and obtain an ideal effect. So it is an effective method for correction of the secondary midface deformities in cleft patients.
Objective To explore the effect of cyclopamine （Cyc） which is the inhibitor of the Hedgehog signaling pathway on portal venous pressure of normal and liver cirrhosis rats, and it’s possible mechanisms. Moreover, to provide the experimental basis of drug efficacy and clinical treatment. Methods Thirty two healthy male SD rats were randomly average divided into four groups：normal control group, normal treatment group, liver cirrhosis control group, and liver cirrhosis treatment group. The liver cirrhosis models of rat were established by using the thioacetamide （TAA） method, which made 0.03% of TAA as the initial water concentration, and then the concentration of TAA in drinking water was adjusted according to the changes of the weekly body weight of rats lasting for twelve weeks. In thirteenth week, intraperitoneal injection of corn oil （0.1 ml/100 g body weight, 1 time/d） were performed lasting for a week in rats of the normal control group and liver cirrhosis control group; intraperitoneal injection of Cyc 〔1 mg （0.1 ml）/100 g body weight, 1 time/d〕were performed lasting for a week in rats of the normal treatment group and liver cirrhosis treatment group. In fourteenth week, the liver function, portal venous pressure （PVP）, and the ration of liver or spleen weight to body weight were detected, the expressions of α-smooth muscle actin （α-SMA） and typeⅠcollagen α1 （Col1α1） of hepatic stellate cell were detected by using immunohistochemistry. Results PVP were （10.7±0.9） and （12.3±1.3） cm H2O （1 cm H2O=0.098 kPa） in normal control group and normal treatment group, respectivly, the latter was higher than the former （t=-2.918，P=0.011）. PVP were （21.8±0.7） and （14.3±1.4） cm H2O in liver cirrhosis control group and liver cirrhosis treatment group, respectivly, the latter was lower than the former（t=13.602，P=0.000）. The expressions of α-SMA and Col1α1 in liver cirrhosis treatment group was lower than the liver cirrhosis control group. There were no significant difference of the liver function and ration of liver or spleen weight to body weight between the treatment group and the control group （P＞0.05）. Conclusion Cyclopamine could signally reduce the PVP of liver cirrhosis rats through reducing the expressions of α-SMA and Col1α1.
摘要:目的:探讨Ⅰ型肾小管酸中毒( RTA) 患者的临床特征及误诊分析。方法: 回顾性分析我院51例Ⅰ型RTA患者的临床资料, 包括临床表现、实验室检查、诊断及误诊情况、并发症及治疗转归等。结果:Ⅰ型RTA 临床表现多样,复杂,无特异性,外院误( 漏)诊率高达64.7%,易并发或伴发多种疾病。 结论: 临床上应提高对Ⅰ型RTA的认识, 做出早期诊断和治疗。Abstract: Objective: To investigate the clinical features and misdiagnosis of distal renal tubular acidosis(RTA), or type 1 RTA. Methods: Data of 51 cases with type 1 RTA were analyzed retrospectively,including clinical features, laboratory examinations,diagnostic errors, complications and therapeutics. Results: The clinical features of type 1 RTA were complicated and were easy to be misdiagnoed.The misdiagnosis rate were as high as 64.7% in other hospitals.Conclusion: Type 1 RTA should be further recognized in clinic in order to make earlier diagnose and treatment.
Neurofibromatosis type 1 (NF1) is an autosomal dominant genetic disease caused by the mutations in the NF1 gene, with an incidence of approximately 1/3 000. Affecting multiple organs and systems throughout the body, NF1 caused a wide variety of clinical symptoms. A comprehensive multidisciplinary diagnostic and treatment model is needed to meet the diverse needs of NF1 patients and improve their quality of life. In recent years, the emergence of targeted therapies has further benefited NF1 patients, and the number of clinical consultations has increased dramatically. However, due to the rarity of the disease itself and insufficient attention previously, the standardized, systematic, and precise diagnosis and treatment model of NF1 still needs to be further improved. In this paper, we reviewed the current status of comprehensive diagnosis and treatment of NF1 in China, combine with our long-term experiences in diagnosis and treatment of this disease. Meanwhile, we propose future directions and several suggestions for the comprehensive diagnosis and treatment model for Chinese NF1 patients.
Objective To investigate the possibility of repairing articular cartilage defects with the mesenchymal stem cells(MSCs) seeded type Ⅰ collagen-glycosaminoglycan(CG) matrices after being cultured with the chondrogenic differentiation medium. Methods The adherent population of MSCs from bone marrow of10 adult dogs were expanded in number to the 3rd passage. MSCs were seeded intothe dehydrothermal treatment (DHT) crosslinked CG matrices; 2×106 cells per 9mm diameter samples were taken. Chondrogenic differentiation was achieved by the induction media for 3 weeks. Cell contractility was evaluated by the measuement of the cell-mediated contraction of the CG matrices with time inculture.The in vitro formation of the cartilage was assessed by an assayemploying immunohistochemical identification of type Ⅱ collagen and by immunohistochemistry to demonstrate smooth muscle actin （SMA）. The cells seededingCGs wereimplanted into cartilage defectsof canine knee joints. Twelve weeks after surgery, the dogs were sacrificed and results were observed. Results There was significant contraction of the MSCsseeded DHT crosslinked CG scaffolds cultured in the cartilage induction medium. After 21 days, the MSCseeded DHT crosslinked matrices were contracted to 64.4%±0.3%; histologically, the pores were found to be compressedandthe contraction coupled with the newly synthesized matrix, transforming the MSCsseeded CG matrix into a solid tissue in most areas. The type Ⅱ collagen staining was positive. The SMA staining was positive when these MSCs were seeded and the contracted CGs were implanted into the cartilage defects of the canine knee joints to repair the cartilage defects. The function of the knee joints recovered and the solid cartilaginous tissue filled the cartilage defects. Conclusion The results demonstrates that MSCs grown in the CG matrices can produce a solid cartilaginous tissuecontaining type Ⅱ collagen after being cultured with the chondrogenic differentiation medium and implanted into cartilage defects. We hypothesize that the following steps can be performed in the chondrogenic process: ①MSCs express SMA, resulting in matrix contraction, thus achieving a required cell density (allowing the cells to operate in a necessary society)； ②Cells interact to form a type Ⅱ collagencontaining extracellular matrix (and cartilaginous tissue)； ③Other factors, suchas an applied mechanical stress, may be required to form a mature cartilage with the normal architecture.