Mycelia glycoproteins from Cordyceps sobolifera ameliorate cyclosporine-induced renal tubule dysfunction in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Cordyceps sorbolifera has been used in Traditional Chinese Medicine for improving the renal function. Cyclosporine A (CsA) is an important immunosuppressive agent in the prevention of renal allograft rejection, but long-term usage of CsA could lead to chronic nephrotoxicity and renal graft failure. The study was aimed to investigate whether the mycelia glycoproteins of Cordyceps sobolifera (CSP) exert prevention effects on CsA-induced nephrotoxicity. MATERIALS AND METHODS: Sprague-Dawley (SD) rats were randomly assigned into four groups (n=6 per group): normal saline (control group), CSP group, CsA group, and CSP-CsA group (CsA combined treatment with CSP). Glomerular and tubular functions were assessed and histological studies were performed. RESULTS: CSP, prepared by hot water extraction, ethanol precipitation and membrane dialysis, was found to be composed of three glycoproteins with average molecular weights of 543, 31, and 6.3 kDa, respectively. CsA impaired urea clearance and creatinine clearance were significantly improved by concomitant administration of CSP. TUNEL histochemical stain revealed that CSP significantly decreased CsA-induced apoptosis in renal tubular cells. The reducing effect of caspase-3 activation by CSP was suggested through the over-expression of the anti-apoptosis protein Bcl-2 in renal tubule cells. In assessment of CSP protection of renal tubule function, we found that CSP restored CsA induced magnesium wasting by increasing the magnesium reabsorption channels TRMP6 and TRMP7. CONCLUSION: The results suggested that CSP had a significant suppressive activity on CsA-induced apoptosis and protective activity against nephron loss possibly via its restoring activity by increasing the magnesium reabsorption channels TRMP6 and TRMP7 on CsA induced magnesium wasting.

Prevention of bone loss by injection of insulin-like growth factor-1 after sciatic neurectomy in rats.

OBJECTIVE: Injection of insulin-like growth factor-1 (IGF-1) can prevent bone loss in sciatic nerve transaction rats. We try to investigate the action mechanism of IGF-1 on bone formation. METHODS: A total of 40 adult male Spragne-Dawley rats were divided into two groups (experimental group and control group) with 20 animals in each. Sciatic neurectomy was performed to model disuse osteoporosis in all rats. IGF-1 was administered in experimental group with the dose of 100 microgramme/kilogram per day for 3 days. Meanwhile, the rats in control group were treated with saline. Bone mineral density was measured by dual-energy X-ray absorptiometry 4 and 6 weeks after neurectomy respectively. Expression of Osterix and Runx2 was determined by reverse transcription-polymerase chain reaction (RT-PCR) assay. RESULTS: There was a significant increase in the bone mineral density of experimental group compared with control group. There was a significant decrease in the level of receptor activator of nuclear factor-kappaB-ligand but an increase in the level of osteoprotegerin 4 and 6 weeks after neurectomy in the experimental group compared with control one. The expression of Osterix and Runx2 was up-regulated in the bone marrow of experimental group compared with control group. CONCLUSION: IGF-1 can increase bone formation by stimulation of osteoblast number and activity, and reduce bone resorption by restriction of differentiation of osteoclast, suggesting that IGF-1 may improve the therapeutic efficacy for disuse osteoporosis.

Substance P stimulates differentiation of mice osteoblast through up-regulating Osterix expression.

OBJECTIVE: To investigate the molecular pathway of substance P (SP) to induce osteoblastic differentiation. METHODS: Mesenchymal stem cells were isolated and cultured. The cultures were divided into four groups with Group A (control group) cultured without any factors, Group B cultured with SP, Group C cultured with SP and SP receptor neurokinin-1 (NK1) antagonist, and Group D cultured with SP NK1 antagonist respectively to induce osteoblastic cells differentiation. Osterix gene expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) for three times after 1-2 weeks of cultivation and the results were analyzed by one-way analysis of variance (ANOVA). RESULTS: The log phase of bone marrow stromal cells appeared at 4-6 days. ALP staining revealed that the majority of cells, more than 95%, were positive and small blue-purple granules were found in the cytoplasm. And Group B, treated with SP, showed a higher level of ALP activity than the other three groups. Meanwhile, RT-PCR found that Osterix expression in Group B was obviously up-regulated, compared with other groups. But Osterix expression in Group D had no remarkable differences, compared with the controls. CONCLUSIONS: SP can up-regulate Osterix gene expression to stimulate differentiation of mesenchymal stem cells into osteoblastic cells at the final stage. The regulatory effect of SP on Osterix expression was dependant on SP NK1 receptors.

[Effect of Xianlong granules on immunological function in rats of adjuvant arthritis].

OBJECTIVE: To study the effect of Xianlong granules (XLG) on immunological function in the rat of adjuvant arthritis (AA). METHOD: Rats were randomly divided into normal group, AA model group, prednisone group and low, middle and high dose XLG groups, 10 rats in each group. All rats were treated by intragastric administration from the 18 days after arthritis was induced by the complete Freud's adjuvant and the effect of XLG on toes swelling was observed. On the 30th days after modeling, proliferation of the splenic and thymic lymphocytes, and IgG secreted by splenocytes were detected respectively by MTT assay and ELISA. RESULT: Compared with the model group, both the high and middle dose XLG groups had significant therapeutic effects on toes dwelling in the rat of AA (P < 0.05 or P < 0.01); The low, middle and high dose XLG groups strengthened the PHAM-inhibited proliferation of splenic lymphocytes (P < 0.05), and inhibited the PHAM-augmented proliferation of thymic lymphocytes (P < 0.05); XLG did not significantly effect on IgG level secreted by splenocytes in rats of AA. CONCLUSION: XLG can cure toes swelling in rats of AA, which is related with regulation of the abnormal immunlological function.

[Effect of arnebia root oil in promoting the tissue recovery of surface of wound and basic fibroblast growth factor (bFGF) mRNA expression].

OBJECTIVE: To explore the molecular biological mechanism of Arnebia Root oil (AO) in promoting the recovery of surface of wound by observing basic fibroblast growth factor (bFGF) mRNA expression in the wound tissue and healing rate of the wound. METHODS: Patients in the observed group with their wound treated by AO and those in the control group treated by petrolatum gauze. The wound surface healing rate was estimated and bFGF mRNA expression was observed by RT-PCR. RESULTS: Endogenous bFGF mRNA expression existed in the wound surface of both groups, but its level in the observed group at any time point was obviously higher than that in the control group respectively, with significant difference in comparison of the gray density between the two groups (P < 0.05). The wound surface healing rate kept abreast with bFGF mRNA expression in wound tissues, so it was significantly higher in the observed group than that in the control group (P < 0.05). GAPDH gene, which was taken as a parameter for internal reference, expressed with a certain amount unchanged in different periods of healing (P > 0.05 ). CONCLUSION: AO shows obviously promotive action on bFGF, an important regulatory factor on wound healing, it might complete the recovery process by stimulating the increase of bFGF.

[Effects of bushen huoxue recipe on proliferation, differentiation and mineralization of osteoblasts in rats].

OBJECTIVE: To observe the effects of bushen huoxue recipe (BHR) on cell proliferation, differentiation, maturation and mineralization in osteoblasts of rats cultured in vitro. METHODS: Osteoblasts from cranium of newborn SD rats were cultured by collagenase method. MTT, PNPP and ARS were used to observe the proliferation, activity of alkaline phosphatase (ALP) and the formation of mineral node of cultured osteoblasts effected by different concentrations of BHR at different time points (24, 48, 72 hrs after medication). RESULTS: Different concentrations of BHR could enhance the cell proliferation rate and ALP activity, the effects of BHR in high and moderate concentration were more significant in enhancing the cell proliferation rate (P < 0.01). And the number of mineral node of cultured osteoblasts treated by high and moderate concentration BHR was much more than that of the untreated osteoblasts (P < 0.01). CONCLUSION: BHR could promote the proliferation, differentiation, and mineralization of the osteoblasts.

[Experimental study of the effects of puerarin on biological characters of osteoblasts in vitro].

OBJECTIVE: To observe the effect of puerarin on cell proliferation, differentiation, maturation and mineralization in cultured rat osteoblasts. METHOD: Osteoblasts from craniums of newly born SD rats were cultured in vitro. MTT, PNPP and ARS were used to observe the proliferation, activity of ALP and the number of mineral node of cultured osteoblasts in vitro. RESULT: It was found that puerarin had the effect on stimulating cell proliferation, activity of ALP and the number of mineral node of cultured osteoblasts (P < 0.01 or P < 0.05). CONCLUSION: Puerarin can promote proliferation, differentiation, maturation and mineralization of the osteoblasts in vitro.

[The expression of c-fos and transmitter calcitonin gene-related peptide in the chronic compressive injury of the nerve root].

OBJECTIVES: To investigate the significance of c-fos oncogene morphogenetic protein's locational expression, and the correlativity between nerve transmitters calcitonin gene-related peptide (CGRP) expression and nerve root's functional change using the animal model of the chronic compressive injury in the nerve root. METHODS: The animal model of chronic compressive injury of the nerve root was established by transplanting autogenous cancellous bone into the intervertebral foramen. During different injury phase (1, 2, 4, 8, 12, 24 weeks after operation), the functional status of the nerve root was determined under the monitoring of evoked potential, and the expression changes of c-fos oncogene morphogenetic protein and nerve transmitter CGRP were detected using in situ hybridization technique and their expression intensity was determined using automatic image analytic instrument respectively. RESULTS: One week after operation, the c-fos expression strengthened in both anterior and posterior root fiber obviously. Two to four weeks after operation, the expression of the posterior root fiber weakened than the anterior root fiber. After 12 weeks, the anterior root fiber expression turned down obviously, however the posterior root fiber expression backed up slightly compared with that of the 8 weeks. By the time of 24 weeks after operation, the expression enhancement in all roots disappeared. CGRP expression increased obviously at the site of compressive axon of both anterior and posterior root. The expression of the posterior root axon and ganglion cell was higher than that of the anterior root axon. CGRP expression was diminished in the second week than the first week, and that was especially obvious in the posterior root and ganglion cell. But 4 weeks after operation, the expression enhanced once more, and that was more obvious inside the anterior root axon. Eight weeks after operation, the expression intensity attained the high peak. Twelve weeks after operation, the expression started the slow-moving descent. CONCLUSIONS: The expression of c-fos gene protein is beneficial to localize the damaged part of certain nerve. During chronic injury, the degeneration of posterior root sensory fiber is earlier than the anterior root motor fiber. The expression of CGRP strengthened when the nerve fiber degenerated by the harmful stimulation, and the expression intensity is positively related with pain. That suggests when the nervous tissue is hurt, the information of warning and regulation should be sent out to our body.

Cloning and characterization of metallothionein gene in ayu Plecoglossus altivelis.

Metallothionein (MT) has been used widely as a potential molecular marker to detect the deleterious effects of heavy metals in aquatic ecosystem. Here we exposed ayu, Plecoglossus altivelis, to zinc (Zn) and tested the distribution as well as the induction of MT in various tissues such as liver, kidney, intestine and stomach. MT induction was significant in liver tissue, followed by kidney and intestine, whereas no induction was detected in stomach. The gene encoding ayu MT was successfully cloned and characterized. Complete nucleotide sequencing and analysis of the 4.5 kb DNA fragment containing the ayu MT gene revealed that the gene has three exons interrupted by two introns, a 5'-flanking region of about 2.5 kb and about 1.6 kb of 3'-flanking region. In grouper heart and kidney cells, the 2.5 kb promoter containing eight metal responsive elements (MREs), two hepatic nuclear factor 5 responsive elements (HNF5REs) and one cAMP responsive element (CRE) had the highest reporter activity.

Comprehensive treatment for old patients with hip fractures.

OBJECTIVE: To observe the effect of comprehensive treatment for hip fracture in old people. METHODS: Three hundred and seventy-two old patients with hip fractures were randomly divided into two groups, Group A and Group B. Cases in Group A were treated only by operations. Cases in Group B received comprehensive treatment. The Singh Indexes of both uninjured and injured femoral necks were used to judge the osteoporosis levels before operation and one year after the operation. The function of injured hip joints was evaluated one year postoperatively. RESULTS: Complications occurred in 36.56% of the cases in Group A and 5.91% of Group B. One year postoperatively, the Singh Index degree distributions of both uninjured and injured femoral necks in Group A had no significant difference compared with those before the operation (P>0.05). In Group B, there was significant difference between one year postoperatively and before operation, and the Singh Index one year after the operation showed better result than that before operation (P<0.05). One year after operation, there was significant difference in the function of injured hip joints between Group A and Group B (P<0.05). CONCLUSIONS: Hip fracture in old people should be treated comprehensively according to its internal characteristics, osteoporosis.