Tendon regeneration induced by umbilical cord graft in a rabbit tendon defect model

Whether tendon regeneration can be induced using the umbilical cord as a whole‐graft structure is unknown. In this study, we explored the potential for tendon regeneration induction using an umbilical cord graft in a rabbit model of patella tendon defects. In 52 of 54 New Zealand White rabbits, the central third of the patella tendons of both hind legs was removed to create tendon defects. The rabbits were randomly divided into four groups, nonfilling (empty defect), refilling (defect refilled with resected tendon portion), Wharton's jelly (WJ) outside (WJO; defect filled with umbilical cord graft, WJ side facing outward), and WJ inside (WJI; same as WJO with WJ side facing inward) groups. Four rabbits from WJO and WJI groups were sacrificed for human CD 105 evaluation 1 month after surgery. Further histological, biomechanical, and gene expression analyses were performed at 3 and 6 months after surgery. The untreated patella tendons in the remaining two rabbits were harvested as normal biomechanical controls. Histological evaluation showed that the formed tissue structure fibers in the tendon defect area were much denser and more mature in the WJI group than in all other groups. Biomechanical testing showed that the failure load of the final tissue structure was the highest in the WJI group. Real‐time polymerase chain reaction indicated that the expression of most tendon‐related genes was upregulated in the WJI group at 6 months after surgery. We concluded that umbilical cord grafting induces effective tendon regeneration, particularly when the WJ side faces inward.     

Treatment of urethral stones by retrograde manipulation and extracorporeal shock wave lithotripsy

Objective To assess the effect of retrograde manipulation and extracorporeal shock wave lithotripsy (ESWL) as a monotherapy for urethral stones that are not associated with urethral strictures. Patients and methods Between August 1993 and January 1995, 34 male patients (mean age 38.7 years, range 7–55) presented with urethral stones. No patient had a suggested or past history of urethral stricture. Lidocaine jelly (2%) was instilled and retained inside the urethra for 5 min. A 16 F urethral catheter was advanced gently to push the stone back to the urinary bladder. Twenty patients had ESWL of their stones in the bladder, using a Storz Modulith SL20, in the prone position. Tilting the patient about 15° towards the side with the stone minimized movement of the stone during fragmentation. In-line co-axial echography (3.5 MHz) and intermittent pulsed fluoroscopy were used to monitor stone fragmentation. In situ fragmentation of posterior urethral stones was not possible because localization was difficult and the treatment was painful. Thirteen patients had cysto-urethroscopy and mechanical cystolitholapaxy under general anaesthesia. Results Stones impacted in the posterior urethra in 31 (91%) patients and in the anterior urethra in three (9%) patients. Stones ranged in size from 7 to 25 mm. One patient expelled an anterior urethral stone after the instillation of 2% lidocaine jelly. The urethral stones were pushed back to the bladder without complication in the remaining 33 patients. All 20 patients except one had their stones fragmented by ESWL in one session. The mean number of shock waves was 3600 ± 1480 (range 1200–6000) and the generator voltage ranged between 5 kV (560 bar) to 8 kV (940 bar). No patient in the ESWL group required anaesthesia or analgesia. Thirteen patients had successful mechanical cystolitholapaxy with no complications. Conclusion Both endoscopic lithotripsy and ESWL of urethral stones are safe and effective. However, transurethral lithotripsy requires general anaesthesia and carries a risk of bladder and urethral trauma. This study demonstrated that, in the absence of urethral stricture, urethral stones can be pushed back safely to the urinary bladder and fragmented effectively by ESWL. The success of the treatment depends on adequate anaesthesia of the urethra before inserting the urethral catheter. We propose that this new technique should be considered before resorting to endoscopic or surgical management of urethral stones, particularly in children.     

蜂王浆冻干粉对实验性高脂血症及血栓形成的影响

用蜂王浆冻干粉每日每公斤体重７００ｍｇ喂饲实验性高脂血症模型大鼠６周，结果发现，蜂王浆冻干粉能降低大鼠血清胆固醇含量，提高高密度脂蛋白胆固醇含量。同时，蜂王浆组大鼠的红细胞变能力增强，血浆纤维蛋白原含量下降，体外血栓指标优于对照组大鼠，差异有统计学意义。提示蜂王浆冻干粉具有防治高脂血症和改善血液高凝状态的作用。     

蚁王浆对慢性肝炎食疗效果的初步观察

本文报告采用蚊王浆食疗33例慢性肝炎患者。临床近期观察结果显示,食疗组治愈8例,好转20例,无效5例。对照组治愈4例,好转9例,无效7例。总有效率前者为84.8％,后者为65.0％(P<0.05)。结果提示,蚊王浆具有缓解肝炎症状,改善肝功能的作用,部分患者乙肝病毒复制指标好转。     

不同消化方式对脐带华通胶间充质干细胞提取的影响

目的探讨使用不同酶浓度、消化不同时间及不同浓度胎牛血清的培养基稀释对消化脐带华通胶后提取培养间充质干细胞的影响。方法无菌条件下取出正常剖腹产胎儿脐带,剔除动脉、静脉及外膜,取其之间的胶状物,剪成1 mm3及更小的块状,分别加入0.1%和0.2%的Ⅱ型胶原酶,分为A、B两组,37℃水浴消化。每一组消化时间分别为4、81、2、16、202、4、28、323、6、40 h共10个消化时间点,每一个消化时间点又分为两组,分别为A1、A2组和B1、B2组。消化后的黏稠细胞悬液用完全DMEM稀释,并加入特级胎牛血清至浓度为10%（A1、B1）和20%（A2、B2）,充分混匀。悬液移至75 cm2培养瓶,15 ml/瓶,37℃、5%CO2、饱和湿度培养,3 d换液。通过细胞计数观察原代细胞贴壁生长的最早时间和80%细胞铺满瓶底的时间和生长活力。结果 0.2%的Ⅱ型胶原酶消化脐带华通胶在消化时间为20h,且稀释血清浓度为20%时,获得的细胞悬液在培养36 h即有细胞贴壁,培养6 d细胞贴壁达到80%,其余各组细胞生长情况各不相等,均弱于此组。其中消化时间在8 h以内和32 h以上时不论使用多少的酶浓度和稀释时使用多少的血清浓度,细胞均不生长。结论在使用胶原酶消化脐带华通胶提取间充质干细胞的过程中,使用0.2%Ⅱ型胶原酶、消化16～24 h、稀释培养时加入至20%的血清对干细胞的提取效果最佳。     

Use of hybrid chitosan membranes and human mesenchymal stem cells from the Wharton jelly of umbilical cord for promoting nerve regeneration in an axonotmesis rat model

Many studies have been dedicated to the development of scaffolds for improving post-traumatic nerve regeneration. The goal of this study was to assess the effect on nerve regeneration, associating a hybrid chitosan membrane with non-differentiated human mesenchymal stem cells isolated from Wharton’s jelly of umbilical cord, in peripheral nerve reconstruction after crush injury. Chromosome analysis on human mesenchymal stem cell line from Wharton’s jelly was carried out and no structural alterations were found in metaphase. Chitosan membranes were previously tested in vitro, to assess their ability in supporting human mesenchymal stem cell survival, expansion, and differentiation. For the in vivo testing, Sasco Sprague adult rats were divided in 4 groups of 6 or 7 animals each:Group 1, sciatic axonotmesis injury without any other intervention (Group 1-Crush); Group 2, the axonotmesis lesion of 3 mm was infiltrated with a suspension of 1 250-1 500 human mesenchymal stem cells (total volume of 50 μL) (Group 2-CrushCell); Group 3, axonotmesis lesion of 3 mm was enwrapped with a chitosan type III membrane covered with a monolayer of non-differentiated human mesenchymal stem cells (Group 3-CrushChitIIICell) and Group 4, axonotmesis lesion of 3 mm was enwrapped with a chitosan type III membrane (Group 4-CrushChitIII). Motor and sensory functional recovery was evaluated throughout a healing period of 12 weeks using sciatic functional index, static sciatic index, extensor postural thrust, and withdrawal reflex latency. Stereological analysis was carried out on regenerated nerve fibers. Results showed that infiltration of human mesenchymal stem cells, or the combination of chitosan membrane enwrapment and human mesenchymal stem cell enrichment after nerve crush injury provide a slight advantage to post-traumatic nerve regeneration. Results obtained with chitosan type III membrane alone confirmed that they significantly improve post-traumatic axonal regrowth and may represent a very promising clinical tool in peripheral nerve reconstructive surgery. Yet, umbilical cord human mesenchymal stem cells, that can be expanded in culture and induced toform several different types of cells, may prove, in future experiments, to be a new source of cells for cell therapy, including targets such as peripheral nerve and muscle.     

Rheological Characterization of an Acetaminophen Jelly

The aim of this study was to prepare an inclusion complex of acetaminophen and β-cyclodextrin (molar ratio of 1:1). A jelly with inclusion complexes formed by kneading was prepared. The formation of inclusion complexes was assessed by powder X-ray diffraction patterns and Fourier transform-infrared spectroscopy. Jellies were prepared with xanthan gum, gelatin, and κ-carrageenan. The concentration of each jelling agent was 0.5, 1.0, and 1.5% w/v. Viscoelasticity and dissolution characteristics were determined and osmometry was performed. PGWater^™, a commercial jelly for fluid replacement, served as a reference for viscoelastic characteristics and dissolution. Powder X-ray diffraction measurement revealed a different diffraction pattern for the kneading than for acetaminophen and β-cyclodextrin. Fourier transform-infrared spectroscopy revealed an absorption peak (at around 1655 cm⁻¹) due to the carbonyl group and benzene ring (at around 1610 cm⁻¹) of acetaminophen. In contrast, the kneaded mixture (1:1) had a shift in the absorption peak due to the carbonyl group (at around 1650 cm⁻¹) in acetaminophen''s molecular structure, and the formation of an inclusion complex was noted. The viscosity of xanthan gum-1.0, gelatin-1.5, and carrageenan-0.5 resembled the viscoelasticity of PGWater^™. The acetaminophen in gelatin-1.0 and carrageenan-0.5 had dissolution behavior similar to that of commercial acetaminophen preparations. The osmolality of jellies prepared in different concentrations ranged from about 20-50 mOsm/kg. Results suggested that carrageenan-0.5 could serve as a useful jelly vehicle for acetaminophen.     

Efficacy of Royal Jelly on Methotrexate-Induced Systemic Oxidative Stress and Damage to Small Intestine in Rats

The aim of this present study is to investigate the mucositis caused by methotrexate (MTX), as well as whether the application of royal jelly (RJ) has a protective effect on oxidative stress. This present study included six groups each consisted of 12 Wistar rats. Distilled water (po: peroral) was given to the 1^st group as placebo for 10 days and MTX (20 mg/kg, intraperitoneal: ip) on the 7^th day. The 2^nd group received RJ (50mg/kg, po) for 10 days and normal saline (NS) instead of MTX. RJ (50mg/kg) was given to the 3^rd group for 10 days and MTX on the 7^th day. The 4^th group received RJ (100 mg/kg, po) for 10 days and NS was given intraperitoneally. RJ (100mg/kg) was given to the 5^th group for 10 days and a single dose of MTX. Distilled water was given to the 6^th (control) group for 10 days and intraperitoneal NS on the 7^th day. Malondialdehyde (MDA), glutathione peroxidase and superoxide dismutase were analyzed in blood samples on the 11^th day. Morphological and histopathological changes were examined in the intestinal tissue samples. Villus length and mucosal thickness, as well as the villus length/crypt ratio, were significantly decreased with MTX administration, and the semi-quantitative histological evaluation (SQHE) score was measured high (p<0.001). In addition, a decrease in the antioxidant parameters and an increase in the MDA levels were identified. The villus length and SQHE were significantly different in the groups receiving RJ (p<0.001) as compared to the MTX group. Although RJ addition had no effect on the decreased mucosal thickness and villus/crypt ratio in MTX groups, it caused an improvement in the antioxidant levels and a remarkable decrease in MDA levels. Adding RJ has a decreasing effect on the MTX-induced intestinal damage and it has a suppressive effect on MTX-induced oxidative stress by means of increasing antioxidant enzyme activity and decreasing lipid peroxidation.     

The stimulating effects of polyphenol and protein fractions from jelly fig (Ficus awkeotsang Makino) achenes against proliferation of leukemia cells

This study aimed to investigate the direct and immune-stimulated antiproliferative activities of jelly fig achenes fractions including pectinesterase inhibitors, crude polyphenols extract, and purified polyphenols extract (PP). Beside the measurement of cell viability of U937, the quantity of cytokines in conditioned medium and morphologic changes in leukemia were observed. After surveying all fractions in jelly fig, the obtained fractions of polyphenol exhibited the highest stimulating effects and directly cytotoxic effects against leukemia with the lowest effect found in protein fractions. The leukemia treated by our PP fraction showed dose-dependent response between the concentration and G2/M cell numbers of the U937 cells. The PP fraction had more pronounced effect on immune-stimulated than direct antiproliferative activities. The finding was also supported by morphological analysis by showing the formation of apoptotic bodies and differentiation from immature U937 cells into mature monocytes/macrophages on cells cultured with PP-conditioned medium. In conclusion, polyphenol fraction of pectinesterase inhibitors from jelly fig showed the immune-stimulated antiproliferative activities against U937 cell.