BACKGROUND: Bushen Huogu decoction can effectively prevent and treat osteoporosis, but the concrete mechanism of pharmacology is still not clear. 25-hydroxy vitamin D3 and 1, 25-dihydroxyvitamin D3 are important coupling factors, which can regulate bone resorption and formation.OBJECTIVE: To investigate the curative effects of Bushen Huogu decoction on the bone mineral density, bone biomechanics, and level of 25-hydroxy vitamin D3 and 1, 25-dihydroxyvitamin D3 in blood serum, liver and kidney in the ovariectomized osteoporotic rats.METHODS: Totally 108 healthy female Sprague-Dawley rats were randomly divided into model group, sham-operated group, andtreatment group. All rats had been ovariectomized to induce estrogen absence and further establish osteoporotic models, except those in sham-operated group. Treatment group of rats were intragastrically administrated with 2 mL Bushen Huogu decoction,twice a day.RESULTS AND CONCLUSION: Compared with model group, the bone mineral density in the rat femur was significantly increased in the treatment group (P < 0.05), the index of maximal stress and maximal loading of the femoral head were also increased (P <0.05). The concentration of 25-hydroxy vitamin D3 and 1, 25-dihydroxyvitamin D3 in blood serum, liver and kidney were significant higher in the treatment group than those in the model group, and the levels were similar with those in sham-operated group (P >0.05). In the early period of estrogenic hormone absence, the Chinese kidney-tonifying drugs could activate bone metabolism, raise bone mineral density and reinforce quality of bone through up-regulating expression of 25-hydroxy vitamin D3 and 1,25-dihydroxyvitamin D3.

Objective:To explore the correlation between the characteristics of contrast-enhanced sonography of intraoperative glioblastoma multiform (GBM) and molecular markers of isocitrate dehydrogenase-1(IDH1).Methods:A retrospective analysis were performed in 30 patients who underwent neurosurgery and pathologically confirmed to be GBM at Beijing Tiantan Hospital from May 2018 to April 2019. All neurosurgical glioblastoma patients after craniotomy underwent conventional ultrasound and contrast-enhanced ultrasound(CEUS) guided navigation. The characteristics of the ultrasound imaging (whether the tumor involves the structure of the corpus callosum, the clarity of the tumor boundary after enhanced ultrasound and whether the tumor has necrotic areas with enhanced ultrasound images) were analyzed. The ratio between tumor necrosis area and whole tumor area (N/W) was measured, and the correlation with IDH1 gene expression was analyzed.Results:There were statistical differences in clarity of tumor boundary after CEUS and tumor necrosis after CEUS between positive IDH1 and negative IDH1 groups(all P<0.05). The positive expression of IDH1 was negatively correlated with the N/W area of the contrast-enhanced ultrasound mode( r=-0.756, P<0.05), suggesting that the expression level of IDH1 gene was negatively correlated with the area of tumor necrosis. Conclusions:Ultrasound contrast agent examination can more accurately distinguish the active proliferation area, hemorrhagic necrosis area and peripheral edema area of glioblastoma. Accurately identifying the extent of tumor necrosis area through ultrasound contrast agent examination can predict expression of IDH1.

Objective To investigate the effects of Dangshen (DS) extract saponins alleviating apoptosis in ischemia-reperfusion injury (I/R) of renal grafts and the mechanism. Methods The I/R injury model in SD rats was established after kidney transplantation. The SD rats were randomly divided into the following 3 groups (n = 20): sham operation group, I/R model group, DS saponin intervention group. Blood urea nitrogen (Bun) and creatinine (Scr) levels were determined at the 24th h after operation; apoptosis index (AI) was detected by using TUNEL method; the expression of Bcl2 and bax mRNA was detected by RT-PCR. Results As compared with the sham operation group,the blood Bun and Scr levels were significantly increased in the I/R model and DS saponin intervention groups (P＜0. 05). As compared with the DS saponin intervention group, the blood Bun and Scr levels were significantly increased in the I/R group (P＜0.05). The AI was significantly increased in the I/R model group and DS saponin intervention group. After DS saponin intervention, the AI was decreased from 40. 28 % in the I/R model group to 28. 45 % in the DS saponin intervention group (P＜0. 05). The expression of Bcl-2 mRNA in DS saponin intervention group and I/R model group was significantly decreased as compared with the sham operation group (P＜0. 05), but that of Bax mRNA was significantly increased in the I/R model group and DS saponin intervention group as compared with the sham operation group (P＜0. 05). After DS saponin intervention, the expression of Bcl-2mRNA was increased from 0. 25 in the I/R model group to 0. 391 (P＜0. 05), and that of Bax mRNA was decreased from 0. 565 in the I/R model group to 0. 473 (P＜0. 05). Conclusion DS extract saponin could significantly alleviate apoptosis in the I/R injury of renal grafts possibly by up-regulating the expression of Bcl-2 mRNA and down-regulating the expression of Bax gene.

Objective To investigate the effect of pretreatment with U75302,antagonist of leukotriene B4 receptor 1 (BLT1),on cisplatin induced acute kidney injury in mice and its immunoregulatory mechanism.Methods Healthy C57BL/6 mice were randomized into four subgroups:1.healthy control group;2.cisplatin group;3.U75302 control group;4.cisplatin + U75302 group,n=6.Group 2 and 4 received intraperitoneal injection of cisplatin (20 mg/kg) on day 0,group 3 and 4received intraperitoneal injection of U75302 (5 μg/mouse) on day 0 and day 2.Mice were sacrificed on the 3rd day and blood and kidney were collected.Renal function and histological changes were estimated,the infiltration of immune cells were determined by flow cytometry,the level of peroxidase (MPO) in kidney were determined by colorimetry,relative expression of TNF-α,IL-1β,CXCL1,CXCL2 were detected by Real-time PCR.Results Compared with healthy control group,levels of BUN,Scr were higher in cisplatin group with serious tubular structural damage.There were more neutrophils,macrophages,CD4+ T lymphocytes,CD8+ T lymphocytes in kidneys of cisplatin group,the level of MPO and relative expression of TNF-α,IL-1β,CXCL1,CXCL2 were also higher in cisplatin group.Compared with cisplatin group,lower BUN [(17.75±1.80) mmol/L vs (42.6±6.66) mmol/L,P ＜0.05],Scr were found in cisplatin+ U75302 group with less tubular structural damage.Meanwhile,U75302 reduced infiltration of neutrophils [(146±13)×103/g vs (296±66) ×103/g,P ＜ 0.05],macrophages [(245± 13)× 103/g vs (420±78)× 103/g,P ＜ 0.05] in the kidney.Levels of MPO [(1.756±0.283) U/g vs (3.308±0.577) U/g,P＜0.05] and relative expression of TNF-α,IL-1β,CXCL1,CXCL2 were also lower.Conclusions BLT1 antagonist U75302 protects mice against AKI induced by cisplatin,and the mechanism is associated with reduced infiltration of inflammatory cells in kidney and the inhibition of kidney inflammation.

OBJECTIVETo investigate the characteristics and incidence of the thoracic deformities in patients with microtia.

METHODSIn Plastic Surgery Hospital, we conducted a retrospective study of the clinical and radiographical data of 300 patients with microtia from March 2013 to October 2014. Pearson χ2 test was used to analyze the relationship among deformities of ribs and spine, as well as microtia.

RESULTSA total of 78 (26.0%) patients were documented with rib deformities, 26 patients (8.7%) had spinal deformities, and 17 patients (5.7% )had both. The incidence of rib deformities in microtia I, II, and III was 7.1% (2/28), 26.7% (62/232) and 35.0% (14/40) respectively. The incidence of spinal deformities in microtia I, II, and III was 3.6% (1/28), 6.5% (15/232) and 25.0% (10/40 respectively. The patients with microtia III were found to have a higher incidence of ribs and spinal deformities than those with microtia II, patients with microtia II were found to have a higher incidence of ribs and spinal deformities than those with microtia I (P < 0.05).

CONCLUSIONSThe incidence of ribs and spinal deformities is high in patients with microtia. The poorer one auricle developed, the higher the incidence of thoracic deformities.

Biomedical Research ; Congenital Microtia ; epidemiology ; Humans ; Incidence ; Retrospective Studies ; Ribs ; abnormalities ; Spine ; abnormalities

Aim To establish a model of the vascular endothelial cell (VEC) injury in SDrats.Methods SD rats were randomly divided into the control and the modelgroups. The model rats were injected with adrenaline diluted to 2. 5 times 0. 05 mg?100 g-1 (tid) for 5 d continously. From the 4th d, they were irritated for 5 min in the0℃ cold-water in the middle between adrenaline injections.The control rats weregiven 0. 9% NS as above. At 6th d, blood samples were taken from carotid arteries ofthe rats and the CEC counts, t - PA、PAI activities, 6-keto-PGF1? concentrations andthe platelet aggregation rate(max) were detected respectively. Results In the modelgroup, as compared with those in the control group, t - PA activity and 6-keto-PGF1?concentration decreased significantly(P

Coptis chinensis is widely used as Chinese medicine herbs and serious soil problems occur after continual cultivation of this medicinal plant. In the preset experiment, fibrous root extract of C. chinensis (REC) was added into soil to study the effect of REC on microbes and enzyme activity in soil. The results showed that both bacteria and actinomycetes decreased by about 2 times in contrast to fungi, which increased by about 3 folds. Phosphorus bacteria, potassium bacteria, azotobacter, ammonia bacteria, and nitrifying bacteria were also reduced significantly by REC, suggesting the inhibition of nitrogen biofixation and supply, mobilization of phosphorus and potassium, ad plant growth promotion as REC added into soil. There were multiple influences of REC on soil enzyme activities. Invertase activity was stimulated, while urease was inhibited and dehydrogenase unchanged by REC, indicating the interference of biochemical reactions in soil. In addition, type and total content of phosphorus lipid fatty acids (PLFAs) , the signature of microbes, decreased while the ratio of bacterium to fungus PLFAs increased as REC increased in soil, which suggested that fungi increased relatively with bacteria decreased thereby leading to easy occurrence of crop fungus diseases following cultivation of C. chinensis. The decrease in diversity and evenness indexes of microbial community in soil by REC indicated soil ecosystem deterioration and reduction of microbial groups and densities in soil. Therefore, allelopathic chemicals released from the roots of C. chinensis could change microbial community structure and resulted in serious soil problems by continual cropping of this medicinal plant.
Coptis ; Ecosystem ; Plant Extracts ; pharmacology ; Plant Roots ; Soil ; chemistry ; Soil Microbiology

BACKGROUND:The cryopreservation of single tissue has achieved great advancement and is gradual y applied in clinics. However, the cryopreservation of complex tissue is rarely reported.
OBJECTIVE:To investigate the morphological change in rabbit limb tissue after replantation through different rewarming methods, find the best rewarming methods of compound textured blood vessels, and provide theoretical basis for the feasibility of limb replantation after long-term cryopreservation.
METHODS:Thirty New Zealand white rabbits were randomly divided into control group, slow freezing-slow thawing group, and slow freezing-rapid thawing group. The right posterior limbs of al the rabbits were cut off 1 cm above the knee joint. Except control group, the latter two groups were given limb replantation after thawing, and then the right posterior limb was again cut off after the replanted limbs were survived for 6 hours. For al groups, the histological changes and gross observation in aorta tissue were observed by light microscopy and transmission electron microscope, and the results were analyzed with statistical methods.
RESULTS AND CONCLUSION:In the slow freezing-slow thawing, slow freezing-rapid thawing groups, the pathological changes (gross specimen, light microscope, electron microscope) of rabbit limbs 6 hours after replantation were worse than those in control group. Compared with slow freezing-rapid thawing group, better integrity of endothelial cells and less damage of the organel es were found in slow freezing-slow thawing group. Through deep cryogenic freezing-thawing process, rabbit limb blood vessels can maintain the structural integrity after replantation and survived at 6 hours. Slow freezing-slow thawing is better than slow freezing-rapid thawing for the preservation of severed limbs, providing evidences for the long-term survival fol owing a deep cry ogenic treatment after the severed limb replantation.

Objective To observe the quantity change of autophagosomes in podocytes and expressions of autophagy-related gene Beclin-1 and microtubule-associated protein 1 light chain 3 (LC3) in different pathological stages of idiopathic membranous nephropathy (IMN),and to explore how autophagy is related to podocyte injury,the occurrence of proteinuria and the disease progression in IMN.Methods Clinical data of 26 patients who were diagnosed as IMN (14 IMN stage 1 and 12 IMN stage 2) admitted to Zhejiang Provincial people's Hospital from January 2013 to December 2014 were retrospectively analyzed.Normal renal tissue from 15 cases of kidney neoplasms with nephrectomy was collected as control.The changes of kidney tissue pathology were detected after PAS and PASM staining by light microscope.The autophagosomes of podocyte were detected by transmission electron microscopy.Expressions of Beclin-1 and LC3 protein were detected by immunohistochemistry.Expressions of LC3 and synaptopodin were detected by immunofluorescence.The correlation of autophagosomes and clinical pathologic factors in IMN patiens was analyzed.Results There were fewer autophagosomes of podocytes and lower expression of Beclin-1 and LC3 protein in IMN group than those in control group (P=0.034,P=0.011,P=0.013,respectively).Moreover,these effects were more obvious with the development of IMN.Compared with those in control group,autophagosomes,Beclin-1 and LC3 protien were reduced in IMN stage 2 group (P=0.009,P=0.030,P=0.015);the number of autophagosomes and the expressions of LC3 and Beclin-1 were decreased in IMN stage 1 group as well,however statistically insignificant (P=0.352,P=0.087,P=0.128);Comparisons between IMN stage 2 patients and IMN stage 1 patients shown significant difference in the number of autophagosomes (P=0.030),but no significant difference in expressions of Beclin-1 and LC3 (P=0.355,P=0.181).Autophagosomes number was not correlated with serum creatinine,serum urea nitrogen,24-hour urinary protein and eGFR (all P ＞ 0.05).The expressions of synaptopodin and LC3 protein were lower in IMN group than those in control group.Conclusion Autophagy may contribute to podocyte injury and the production of protein urine in IMN,and may be closely related to the progression of disease.

Background Pterygium is one of the common ocular surface disorders,and the main drugs for pterygium include dexamethasone (DXM),interferon α-2b (IFN-α2b),mitomycin C (MMC),5-fluorouracil (5-FU),cyclosporin A (CsA) and tacrolimus (FKS06).However,the efficacy of these drugs on the fibroblasts from recurrence pterygium is unelucidated.Objective This study was to compare the efficacy of DXM,IFN-o2b,MMC,5-FU,CsA and FK506 on proliferation and apoptosis of recurrent pterygium-derived fibroblasts in vitro.Methods The specimens of recurrence pterygium were collected during surgery in Tianjin Medical University Ophthalmological Hospital from May 2015 to July 2016 under the written informed consent.Fibroblasts were isolated and cultured by explant culture method and identified by immunochemistry.DXM,IFN-α2b,MMC,5-FU,CsA and FK506 were added into the medium for 48 hours,respectively,and the cells cultured without drug were used as the control group.The inhibitory efficiency of different concentrations of DXM,IFN-α2b,MMC,5-FU,CsA and FK506 on the cell proliferation was assayed by cell counting kit-8 (CCK-8),and 50％ inhibiting concentration (IC50) of the drugs was calculated.The cells were treated by the IC50 dose of drugs for 48 hours,and cell apoptotic proportion and cell cycle were assessed by flow cytometry analysis.The expression of proliferating cell nuclear antigen (PCNA) in the cells after treated by drugs was detected by immunochemistry.Results Cultured cells grew well with the fusiform shape and radial arrangement.Vimentine showed the positive expression and keratin was absently expressed in the cells.The IC50 to the cells was (3.5×103±2.83×10-2)mg/L,(6.1×102±3.6×10-3)mg/L,(3.2×10-1±1×10-4)mg/L,(2.2× 101 ± 1.2× 10-3) mg/L,(6.3 × 101 ±2.5 × 10-3) mg/L and (6.0× 101 ± 0.0× 100) mg/L in the DXM,IFN-α2b,MMC,5-FU,CsA and FK506,respectively.In the 48 hours after treated by the IC50 drugs,the apoptotic ratio was (35.00± 3.21)％,(30.37±1.67)％,(26.11±0.75)％,(22.01±0.07)％,(20.95±1.68)％ and (19.85±0.52)％ in the IFN-α2b group,CsA group,MMC group,FK506 group,DXM group and 5-FU group,which was significantly higher than (11.38±2.18) ％ in the control group (all at P＜0.05).The cell proportion of G0/G1 phase,S phase and G2/M phase was (85.64±2.62)％,(5.29±1.56)％ and (2.73-±2.66)％ in the control group,and the cell proportion of G0/G1 phase was reduced,while that of S phase or G2/M phase was considerably increased in various drug groups (all at P＜0.05),with the blocking efficiency of cell cycle was in turn MMC,CsA,5-FU,DXM,IFN-α2b and FK506.The expressional rate of PCNA in the cells was (95.00 ± 2.00) ％,(82.67 ± 5.04) ％,(80.00 ± 2.78) ％,(64.00± 6.55)％,(38.00±3.00)％,(32.00±4.36)％ and (29.67±3.02)％ in the control group,FK506 group,DXM group,5-FU group,IFN-α2b group,CsA group and MMC group,showing a significant difference among the groups (F=25.995,P＜0.01),and the expressional rate of PCNA was significant lower in various drug groups than that in the control group (all at P＜0.05).Conclusions DXM,IFN-α2b,MMC,5-FU,CsA and FK506 are all able to inhibit the proliferation and promote the apoptosis of recurrent pterygium-derived fibroblasts in vitro,and MMC and CsA appear to have a stronger effect.