[Objective]To understand the changes of periprosthetic bone mineral density(BMD) of the proximal femur in patients with failed hip arthroplasty by measuring with dual energy X-ray absorptiometry(DEXA). [Method]Periprosthetic BMD was determined in 9 patients with revision total hip arthroplasty by dual energy X-ray absorptiometry(DEXA),defining regions of interest according to Gruen(1~7),and comparing the operation side with the nonoperation side.Clinic results of the cases were evaluated according to Harris Hip Score(HHS).[Result]The mean HHS was 61 points in 9 patients at this follow-up examination.There was a significant decrease(8.9%~27.6 %) in BMD in every Gruen zone as compared with the nonoperation side.[Conclusion]The bone loss in the proximal femur in patients with failed hip arthroplasty is significant,and the pattern of bone loss around failed hip arthroplasty may be different from the typical remodeling seen after successful hip arthroplasty.

[Objective]To discuss the outcome of acetabular protrusion after total hip arthroplasty. [Method]Fifteen patients(M=7,F=9) having painful acetabular protrusion received total hip arthroplasty with reinforcement of the acetabulum with bone grafts(autogenous or allogeneous bone).The results were reported at 1~4 years(mean,3 years) of follow-up.Of the 15 cases,5 were mild protrusion and 10 were moderate protrusion.The Harris hip-rating and radiographs were taken pre-and postoperatively. [Result]The average Harris hip-rating was improved from 45 points(range,39~60 points) preoperatively to 85 points(range,70~100 points) postoperatively.The average preoperative protrusion of the femoral head medial to the Kohler line was 8.8 mm(6~18 mm).The average postoperative placement of the prosthetic femoral head was 10 mm(6~13 mm) lateral to the Kohler line.None acetabulum prothesis failure or bone grafts absorption was seen.The grafts were well incorporated on radiograms in all patients one year after operation by radiographs. [Conclusion]Good results can be obtained in hips with acetabular protrusion after reconstructed with autogeneous or allogeneous bone grafting and total hip arthroplasty.

Objective To explore the treatment methods of periprosthetic femur fracture after hip arthroplasty.Methods 9 patiens with periprosthetic femur fracture after hip arthroplasty were selected.According to Vancouver classification,there were 1 case in A type,5 cases in B1 type,2 cases in B2 type,1 case in C type.One case were treated by nonoperative method and the other 8 cases were treated by operative method,including 5 cases treated by memory alloy embracing fixator and internal fixation,3 cases treated by long stem prosthetic replacement and iliac bone graft.Results 8 cases were followed up for 8 to 21 months,average 14.2 months.All fractures were united well with good alignment and internal fixation failure except one prosthesis loosing was observed.Conclusion For periprosthetic femur fracture after hip arthroplasty,Vancouver classification methods include the location and stability of the fracture,prosthesis loosening,and the femur in bone mass is importance to the clinical treatment.As to A type fracture,prosthesis is stable,and the conservative treatment can be choosed.As to B1 and B2 type fracture,the aggressive surgical treatment can be choosed based on patients' general condition.If prosthesis loosening after artificial hip arthroplasty,the patients should treated with long stem prosthetic replacement.

Objective To study the genetic diversity of Fritillaria thunbergii,a traditional Chinese herb in Zhejiang Province in China.Methods The genetic diversity of six representational populations of F.thunbergii including 32 individuals was investigated by amplified fragment length polymorphism(AFLP) maker technique.Results The genetic diversity was revealed as follow: the Nei′s genetic diversity index(He) 0.169 0?0.175 7,Shannon′s information index(I) 0.269 8?0.245 3,percentage of polymorphic loci(PPB) was 76.85% at the species level;Ht 0.169 0?0.030 9,and Hs 0.150 8?0.024 0,I 0.233 3?0.261 9, PPB was 50.38% at population level.The genetic differentiation index(Gst) was 0.107 6,Nm 4.147 0.The result of dendrogram of six populations indicated that Dongyang and Yongkang populations shared the minimum genetic distance(0.015 0),they were classified into a group,and Xiangshan and Jinyun populations shared the maximum genetic distance(0.032 4).Conclusion The genetic diversity of F.thunbergii cultivated in Zhejiang Province is very rich,which could ensure the long-term survival of F.thunbergii.But the genetic diversity of F.thunbergii is relatively higher in population levels while lower at the species levels and the degree of genetic differentiation occured among the populations is not significant.The germplasm resources are relatively stable among these six populations.These populations could be used to breed the fine strains of F.thunbergii as the bases.

OBJECTIVE: To observe the effect of wheat-grain moxibustion at "Dazhui" (GV 14), "Zusanli" (ST 36) and "Sanyinjiao" (SP 6) on Wnt/β-catenin signaling pathway in bone marrow cell in mice with bone marrow inhibition, and to explore the possible mechanism of wheat-grain moxibustion in treating bone marrow inhibition. METHODS: Forty-five SPF male CD1(ICR) mice were randomly divided into a blank group, a model group and a wheat-grain moxibustion group, 15 mice in each group. The bone marrow inhibition model was established by intraperitoneal injection of 80 mg/kg of cyclophosphamide (CTX). The mice in the wheat-grain moxibustion group were treated with wheat-grain moxibustion at "Dazhui" (GV 14), "Zusanli" (ST 36) and "Sanyinjiao" (SP 6), 3 moxa cones per acupoint, 30 s per moxa cone, once a day, for 7 consecutive days. The white blood cell count (WBC) was measured before modeling, before intervention and 3, 5 d and 7 d into intervention. After intervention, the general situation of mice was observed; the number of nucleated cells in bone marrow was detected; the serum levels of interleukin-3 (IL-3), interleukin-6 (IL-6) and granulocyte macrophage colony stimulating factor (GM-CSF) were measured by ELISA; the protein and mRNA expression of β-catenin, cyclinD1 and C-Myc in bone marrow cells was measured by Western blot and real-time PCR method. RESULTS: Compared with the blank group, the mice in the model group showed sluggish reaction, unstable gait, decreased body weight, and the WBC, number of nucleated cells in bone marrow as well as serum levels of IL-3, IL-6, GM-CSF were decreased (P<0.01), and the protein and mRNA expression of β-catenin, cyclinD1 and C-Myc was decreased (P<0.01). Compared with the model group, the mice in the wheat-grain moxibustion group showed better general condition, and WBC, the number of nucleated cells in bone marrow as well as serum levels of IL-3, IL-6, GM-CSF were increased (P<0.01, P<0.05), and the protein and mRNA expression of β-catenin, cyclinD1 and C-Myc was increased (P<0.05). CONCLUSION Wheat-grain moxibustion shows therapeutic effect on bone marrow inhibition, and its mechanism may be related to activating Wnt/β-catenin signaling pathway in bone marrow cells, improving bone medullary hematopoiesis microenvironment and promoting bone marrow cell proliferation.
Animals ; Male ; Mice ; beta Catenin/metabolism* ; Bone Marrow/physiopathology* ; Bone Marrow Cells/physiology* ; Granulocyte-Macrophage Colony-Stimulating Factor/metabolism* ; Interleukin-3/metabolism* ; Interleukin-6/metabolism* ; Mice, Inbred ICR ; Moxibustion/methods* ; RNA, Messenger/metabolism* ; Triticum ; Wnt Signaling Pathway ; Hematopoiesis

Objective By using quantum dots-based double-color imaging method to simultaneously label human epidermal growth factor 2 (HER2) on breast cancer cell membrane and the type Ⅳ collagen in the extracellular matrix.To monitor the degree of malignancy of breast cancer and the invasive and metastatic potential.Methods The co-expressions of HER2 and the type Ⅳ collagen in breast cancer of 54 cases were detected and quantified by quantum dots-based double-labeling immunofluorescent histochemistry,and its correlation with clinical pathology parameters and prognosis were analyzed.Results With the increasing HER2 expression level,a progressive decrease in collagen Ⅳ around the cancer nest,the expression of HER2 and the type Ⅳ collagen value was negatively correlated (r =-0.980,P ＜ 0.05) ; the expression of HER2 and collagen Ⅳ was associated with lymph note metastasis,pathological stage (TNM) and disease-free survival(P ＜ 0.05).The expression of HER2 and the type Ⅳ collagen between different age groups,menopause and estrogen receptor (ER) and progesterone receptor (PR) were not significantly different (P ＞ 0.05).Conclusions Quantum dots-based double-color imaging method provides direct observable evidence to support the degradation of HER2 and the type Ⅳ collagen expression,which may help in determining the degree of malignancy and evaluating prognosis.

Objective: To compare the protective effect of Phytolaccae Radix and its processed products on nephropathy induced by doxorubicin (DOX) in rats, and explore its mechanism. Method: A rat model of nephropathy was established by a single tail intravenous injection of DOX hydrochloride. Content of esculentoside A (EsA) in Phytolaccae Radix and its processed products was determined by HPLC-ELSD. Contents of serum total protein (TP), albumin (Alb), urea nitrogen (BUN), serum creatinine (SCr), total cholesterol (TC) and urine protein (UP) were determined by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein expression of transforming growth factor-β (TGF-β) in renal tissue of rats was examined by real-time quantitative polymerase chain reaction (Real-time PCR) and immunohistochemistry. Result: A single intravenous injection of DOX could induce a severe nephrotic syndrome associated with decreased serum TP, Alb and elevated serum BUN, SCr, TC, and a high urinary excretion of protein (P<0.05). Expression of BUN, SCr, TC in serum and UP in urine of model rats could be decreased by Phytolaccae Radix and its processed products to some degree, expression of TP and Alb in serum of model rats could be increased by Phytolaccae Radix and its processed products to some degree, vinegar processed products had the most significant effect. Expression of TGF-β in renal tissue of model group rats was significantly higher than that of blank group (P<0.01), which could be significantly reversed by Phytolaccae Radix processed with vinegar, Phytolaccae Radix boiled with vinegar, Phytolaccae Radix steamed with water(P<0.05, P<0.01). Conclusion: Phytolaccae Radix and its processed products can improve the symptoms of DOX nephropathy model rats in different degrees, among which the vinegar prepared products have the strongest effect, and this effect may be related to the reduction of TGF-β expression in renal tissue.

OBJECTIVE: To explore the effect of "Zhibian" (BL 54)-to-"Shuidao" (ST 28) needle insertion on the ovarian function in the rats with primary ovarian insufficiency (POI) and the potential effect mechanism based on the Fas/FADD/Caspase-8 of death receptor pathway. METHODS: Forty-eight female SD rats were randomly divided into a blank group, a model group, a medication group and an acupuncture group, with 12 rats in each group. Except in the blank group, the rats in the other groups were intraperitoneally injected with cyclophosphamide to establish the POI model. In the acupuncture group, after successful modeling, the intervention was given with "Zhibian" (BL 54)-to- "Shuidao" (ST 28) needle insertion, once daily, 30 min in each intervention; and the duration of intervention was 4 weeks. In the medication group, estradiol valerate tablets were administered intragastrically, 0.09 mg•kg^-1•d^-1, for 4 weeks. The general situation and the estrous cycle of the rats were compared among groups. Using ELISA, the levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E₂) in the serum were detected. HE staining was adopted to observe the morphological changes of ovarian tissue of rats. The protein expression of Fas, FADD and Caspase-8 in ovarian tissue was detected with immunohistochemistry and Western blot. RESULTS: After modeling, except the rats of the blank group, the rats of the other groups had dry fur, lost hair, low spirits, reduced food intake, increased urination and loose stool. After intervention, the stool became regular gradually in the acupuncture group and the medication group. The percentage of estrous cycle disturbance was increased in the rats of the model group when compared with the blank group (P<0.01); in comparison with the model group, the percentages of estrous cycle disturbance were reduced in the acupuncture group and the medication group after intervention (P<0.01). When compared with the blank group, the body mass and E₂ content in the serum were lower (P<0.01), the levels of FSH and LH in the serum and the protein expression levels of Fas, FADD and Caspase-8 were increased (P<0.01) in the model group. Compared with the model group, the body mass and E₂ contents in the serum were higher (P<0.01), the levels of FSH and LH in the serum and the protein expression levels of Fas, FADD and Caspase-8 were reduced (P<0.01) in the acupuncture group and the medication group. CONCLUSION "Zhibian" (BL 54)-to-"Shuidao" (ST 28) needle insertion can effectively improve the ovarian function of POI rats, and its effect mechanism may be related to regulating the serum sex hormone levels, reducing the expression of Fas, FADD and Caspase-8 in ovarian tissue and retarding apoptosis of ovarian cells.
Female ; Animals ; Rats ; Needles ; Signal Transduction ; Receptors, Death Domain/metabolism*

ObjectiveTo observe the regulatory effect of Huangwu Ganfu ointment on transient receptor potential anchor protein 1 （TRPV1） receptor expression， macrophage polarization， and p38 mitogen-activated protein kinase （p38 MAPK） signaling pathway in synovial tissue of knee osteoarthritis （KOA） rats with yang deficiency and cold coagulation syndrome and explore the mechanism of relieving peripheral inflammatory hyperalgesia of KOA. MethodForty-eight male SD rats were randomly divided into blank group， model group， high-dose， middle-dose， and low-dose groups of Huangwu Ganfu ointment （9.3， 4.65， 2.325 g·kg^-1）， and celecoxib group （20.82 mg·kg^-1）. The KOA rat model of yang deficiency and cold coagulation syndrome was established through climate box and swimming for two weeks combined with an injection of sodium iodoacetate （MIA） in the articular cavity. After continuous administration for four weeks， the general condition of rats in each group was observed， and the pain withdrawal threshold （PWT） and joint diameter induced by mechanical stimulation were recorded. The expression levels of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， nerve growth factor （NGF）， and calcitonin gene-related peptide （CGRP） inflammatory factor were detected by enzyme-linked immunosorbent assay （ELISA）， and the histopathological changes of synovial tissue of the knee joint were observed by hematoxylin-eosin （HE） staining. Western blot was used to detect the protein expression of TRPV1， p38 MAPK， and p-p38 MAPK in synovial tissue of the knee joint， and immunofluorescence （IF） was used to evaluate the polarization of M1/M2 macrophages. ResultCompared with that in the blank group， the overall mental state of the model group was worse， and the autonomous activity was decreased. The body mass was lower， and the joint diameter was increased. The X-ray showed that the osteophyte at the edge of the joint proliferated， and the articular surface was obviously rough. The articular cavity was significantly narrowed， and the PWT was significantly decreased （P<0.01）. The contents of IL-1β， TNF-α， CGRP， and NGF in serum and synovium Krenn score increased significantly （P<0.01）. The protein expression of TRPV1 and p-p38 MAPK/p38 MAPK increased significantly （P<0.01）， and the proportion of M1 macrophages and M1/M2 increased （P<0.01）， while the proportion of M2 macrophages decreased （P<0.01）. Compared with model group， the body mass in the low， middle， and high dose groups of Huangwu Ganfu ointment increased to different degrees （P<0.05， P<0.01）. The diameter of the knee joint in the high dose group of Huangwu Ganfu ointment and celecoxib group decreased （P<0.01）. The recovery of PWT in the high and middle dose groups of Huangwu Ganfu ointment groups was more obvious （P<0.05）. The contents of IL-1β and CGRP in the serum of rats in each administration group were significantly decreased （P<0.01）， and the content of serum TNF-α in the celecoxib group and high dose group of Huangwu Ganfu ointment decreased significantly （P<0.05）. The content of serum NGF in the middle dose group of Huangwu Ganfu ointment decreased significantly （P<0.05）， and the synovium Krenn score decreased in the high dose group of Huangwu Ganfu ointment （P<0.05）. In addition， the protein expression of TRPV1 and p-p38 MAPK/p38 MAPK in synovial tissue decreased significantly in all groups of Huangwu Ganfu ointment （P<0.01）. The proportion of M1 macrophages in synovial tissue in the celecoxib group and all groups of Huangwu Ganfu ointment decreased （P<0.01）， and the proportion of M2 macrophages in the high dose group of Huangwu Ganfu ointment increased （P<0.05）. The M1/M2 in the middle and high dose groups of Huangwu Ganfu ointment decreased （P<0.05）. ConclusionHuangwu Ganfu ointment can mediate the polarization of macrophages to reduce the inflammatory reaction of KOA， alleviate the release of inflammatory pain mediators， and lower the protein expression of TRPV1. The mechanism may be related to the p38 MAPK signaling pathway， so as to improve the peripheral hyperalgesia of KOA.

Objective:To explore the neuroprotective effect and mechanism of Buyang Huanwu Tang (BYHWT) on experimental autoimmune encephalomyelitis (EAE) at different stages. Method:The 36 female C57BL/6 mice were immunized subcutaneously with myelin oligodendrocyte glycoprotein peptides (MOG_35-55),then randomly divided into 9, 17, 28 d EAE control group. Each BYHWT group was orally given drugs on the 3^rd day after immunization (50 g·kg^-1·d^-1), and EAE control group was given the same volume of normal saline in the same way once a day for 9, 17 and 28 d after immunization. The effect of BYHWT on EAE mice was observed with internationally accepted clinical score. Brain and spinal cord specimens were collected at 9, 17 and 28 d after immunization. The neuroprotective effect of BYHWT was observed by hematoxylin-eosin(HE)staining and solid blue staining (LFB). The expressions of BDNF and GAP-43 in spinal cord and brain were detected by Western blot. Result:After treatment, BYHWT can significantly inhibit myelitis cell infiltration and alleviate myelin loss. Compared with EAE group, the expression of Nogo-A in the spinal cord of each BYHWT group was significantly down-regulated (P<0.01), and the expression of BDNF in the spinal cord was significantly up-regulated (P<0.05, P<0.01) in the BYHWT group 17 and 28 d group compared with EAE group 17 and 28 d group. Compared with EAE 9, 17 d group, GAP-43 expression was significantly up-regulated in the spinal cord of BYHWT 9, 17 d group (P<0.01). Conclusion:BYHWT can improve the local nerve growth microenvironment and promote the expression of NTFs, reduce the expressions of neuroinhibitory factors, and play a role in neuroprotection.