OBJECTIVE: To study the chemical constituents of ethanol extract from Cirtus aurantium. METHODS: The ethanol extract of C. aurantium was isolated and purified by silica gel column, open ODS column, preparation liquid chromatography, HW-40F gel column and macroporous resin column. The structure of compounds was analyzed and identified according to physicochemical properties and spectral data (mass spectrum, hydrogen spectrum, carbon spectrum). RESULTS: Eight compounds were isolated from ethanol extract of C. aurantium, i. e. Rimboxo (1), Thymidine (2), Uracil (3), Cyclo-(Leu1-Ile2-Ala3-Thr4-Gly5-Thr6-Phe7) (4), Cyclo- (Leu1-Leu2-Pro3-Tyr4-Gly5-Ser6-Pro7) (5), Meranzin hydrate-β-D-glucoside (6), Umbelliferone (7), Linaloyl glucoside (8). CONCLUSIONS: Compound 1, 2, 3, 8 are isolated from Citrus L. for the first time, compound 4, 5, 6 were isolated from C. aurantium for the first time. The study lays the foundation for quality evaluation of C. aurantium.

OBJECTIVETo investigate the feasibility of promoting nerve regeneration by using microcapsulated NGF-expressing cells transplantation.

METHODSThe plasmid pcDNA3. 1 + /NGF, containing rat NGF gene, was transfected into the NIH 3T3 cell by using FuGENE6 transfection reagent. The genetically modified cells expressing NGF gene were enclosed within the alginate-polylysine-alginate (APA) microcapsules and then cultured in vitro. The growth and NGF secretion of the cells were measured periodically. At the same time, these microcapsulated NGF-expressing cells were transplanted into the injured sciatic nerve. The regeneration and functional recovery of the nerve were evaluated in 4 weeks, 8 weeks and 12 weeks after the operation.

RESULTSThe microcapsulated cells had survived and secreted the NGF in three months in vitro. In the group with microcapsulated NGF-expressing cells, the number of the regenerated axons was in large and the nerve fibers were arranged regularly. Compared to other groups, there was less scar , edema and monocytes found at the stoma in the goup. The moter nerve conductive velocity, nerve muscle-action potential and SFI were improved.

CONCLUSIONSThe microcapsulated NGF-expressing cells could significantly enhance the nerve regeneration and reduce inflammatory response of xenograft.

Animals ; Cell Transplantation ; Female ; Male ; Mice ; NIH 3T3 Cells ; Nerve Growth Factor ; biosynthesis ; genetics ; Nerve Regeneration ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo analysis the early complications of tibial fracture and its related factors, and propose a solution.

METHODSFrom December 2003 to December 2013,38 patients with early complications of tibial plateau fracture after operation were retrospectively analyzed. There were 35 males and 3 females, aged from 37 to 69 years old (averaged 42.3 years). According to Schatzker classification, 3 cases were classified as type II, 2 cases as type III, 2 cases as type IV, 19 cases as type V, 12 cases as type VI. The intervals between injury and operation ranged from 9 hours to 9 days, 26 cases within 3 days. Fifteen cases were treated with internal fixation of plates and 23 were treated by plate fixation and bone transplantation. Early complications included skin necrosis in 15 cases, infection in 6 cases, osteofascial compartment syndrome in 3 cases, common peroneal nerve injury in 2 cases, the superficial peroneal nerve injury in 3 cases, popliteal artery injury in 2 cases, loss of reduction in 7 cases.

RESULTSThe wound of 14 cases healed at the first stage and 24 cases healed delay. Hospitalization days ranged from 7 to 67 days (averaged 25.6 days). All patients were followed up for 12 to 36 months with an average of 16.4 months. The fracture healing time ranged from 3 to 9 months (averaged 6.9 months). According to Merchant knee function evaluation criteria, the results were excellent in 19 cases, good in 12, fair in 5 and poor in 2.

CONCLUSIONEarly complications of tibial fracture after operation is closely associated with the severe fracture complexity and related with preoperative preparation, surgical timing, operation incision selection and surgical technique. Early detection and timely processing reduce damage.

Adult ; Aged ; Female ; Humans ; Length of Stay ; Male ; Middle Aged ; Postoperative Complications ; therapy ; Tibial Fractures ; surgery

OBJECTIVETo investigate the effects of transforming growth factor-beta1 (TGF-beta1) gene therapy on bone defect and bone rarefaction around endosseous implant.

METHODSThe primary cultured bone marrow derived stroma cells (BMSCs) was transfected by plasmid pCDNA3.1(+) -TGF-beta1, and was adhered with polylactic-co-glycolic acid (PLGA) for constructing TGF-beta1 gene-modified artificial bone. The model of rats with placed titanium implants in the proximal metaphyses of the tibiae after ovariectomy was made. The TGF-beta1 gene-modified artificial bone (experimental group), BMSCs-PLGA compound artificial bone (control group) and nothing (blank control group) were placed in the bone defect around implant. The tibiae were examined by decalcified sections with immunohistochemical method and histological analysis methods at intervals of 4 and 8 weeks after implant surgery in order to detect the expression of TGF-beta1 in new bone adjacent to the implant and the healing of the bone defect around the implant.

RESULTSThe expression level of TGF-beta1 of experimental group was higher than that of control group and blank control group at the 4th week. The histological analysis indicated that the gene-modified artificial bone had stronger osetogenic potential than others.

CONCLUSIONTGF-beta1 gene-modified artificial bone promotes the repair of the bone defect around titanium implants in osteoporotic rats.

Animals ; Bone and Bones ; Cells, Cultured ; Dental Implantation, Endosseous ; Dental Implants ; Female ; Genetic Therapy ; Prostheses and Implants ; Rats ; Stromal Cells ; Titanium ; Transfection ; Transforming Growth Factor beta1

OBJECTIVETo investigate the impact of cement distribution index on the occurrence of refracture in the adjacent segments after percutaneous vertebroplasty.

METHODSThis retrospective analysis was conducted among 143 patients who received percutaneous vertebroplasty for osteoporotic vertebral compression fracture between April, 2011 and April, 2014. Of the 134 patients with complete follow-up data, 18 had adjacent segment fracture within 1 year following the surgeries (re-fracture group), and 116 patients without new fracture served as the control group. All the patients underwent X-ray examinations after the surgery and according to the position and shape, the cement in the vertebrae were classified into 5 types (I to V), and the volume-cubage index was computed based on the cement volume and vertebral cubage. Age, gender, bone mineral density (BMD), cement distribution index, volume-cubage index, and cement leakage were evaluated in the 2 groups, and the variables with significant differences between the 2 groups were analyzed in Logistic regression analysis.

RESULTSBMD was significantly lower and the rate of cement leakage was significantly higher in the re-fracture group than in the control group (P<0.05). Significant difference was found in cement distribution index between the 2 groups (P<0.05) but not in age, gender, cement volume or volume-cubage index (P>0.05). Logistic regression analysis indicated that BMD, cement leakage and cement distribution index all significantly affected the occurrence of adjacent vertebral fractures following percutaneous vertebroplasty.

CONCLUSIONA low BMD, cement leakage and a low cement distribution index are all risks factor of adjacent vertebral fracture after percutaneous vertebroplasty.

OBJECTIVE：To sepa rate and identify the phenols compounds in Aurantii Fructus Immaturus ，and to provide reference for basic research of its effective substances. METHODS ：The phenols compounds were isolated and separated by silica gel，HW-40F gel column ，ODS reversed column and preparation HPLC. Their physicochemical properties and spectral data were used to identify the structures. RESULTS & CONCLUSIONS ：Totally 12 compounds were isolated from Aurantii Fructus Immaturus，identified as 6′-O-trans- cinnamoyl- 3，5-dihydroxyphenyl β-D-glucopyranoside（compound 1），methyl 3-（2′，4′- dihydroxy phenyl ）propionate（compound 2），phloroglucinol（compound 3），aurantiside A （compound 4），5，7，4′-trihydroxy-8， 3′-dimethoxyflavone-3-O-6″-（3-hydroxyl-3-methylglutaroyl） β-D-glucopyranoside （compound 5）， aromadendrin-7-O-β-D- glucopyranside（compound 6），hesperidin-7-O-β-D-glucoside（compound 7），naringin（compound 8），hesperidin（compound 9）， narirutin （compound 10），neoeriocitrin （compound 11），eriocitrin （compound 12）. Among them ，compound 1 is a new compound，and compounds 2 and 3 are isolated from Critus for the first time.

Purpose To investigate the value of application of D2-40/CD34-CK cocktail antibodies by double immunohistochemical staining for assessment of lymphovascular invasion (LVI) and to determine its prognostic significance in colorectal cancer with insufficient lymph node harvest. Methods Specimens from 133 cases of colorectal cancer with less than 12 lymph nodes were selected. HE staining and double immunohistochemical staining of the cocktail antibodies were performed to compare the difference of the two methods in screening for LVI. The The relationship between LVI confirmed by cocktail antibody immunohistochemical staining and clinicopathological characteristics and overall survival (OS) of patients was analyzed. Results (1) The detection rates of cocktail antibody double immunohistochemical staining and HE staining for LVI were 42.9％ (57/133) and 21.8％ (29/133) with statistically significant difference (P < 0.001). (2) The presence of LVI confirmed by double staining was significantly associated with Dukes staging, depth of invasion, clinical stages, lymph node metastasis and tumor budding (P < 0.05). (3) The presence of LVI, the location and extent of LVI, and the number of tumor cells in thrombus ≥5.5 for cases with LVI ≤2 clusters, were significantly associated with OS (P < 0.05). Conclusion D2-40/CD34-CK cocktail antibodies double staining is superior to routine HE staining in assessing LVI. LVI is intimately associated with tumor stage, lymph nodes metastasis and tumor budding, and it is an independent prognostic factor for CRC patients. It should be a supplementary examination for these patients with insufficient lymph node harvest.

OBJECTIVEEstablishing a rabbit model of vertebral artery type of cervical spondylosis(CSA) and to observe the characteristics of timeliness in improving the blood flow of vertebral artery by massage, and discusse the material basis of this timeliness based on NPY and ET-1.

METHODSFifty New Zealand healthy and white rabbits, 6-month-old, the body mass of (2.0±0.5) kg, with half males and half females, were randomly divided into blank group, model group, three massage groups(including massage for 10 min, 20 min, and 30 min group by random number table), 10 rabbits in each group. In addition to the blank group, CSA rabbit model was made by injection of sclerosing agent in other groups. The rabbits of massage for 10 min, 20 min, and 30 min groups received the massage therapy of corresponding duration, one times a day, continuous 10 days. The blood flow of vertebral artery in each group was detected by PeriFlux5000 laser doppler, and the contents of NPY and ET-1 in serum were detected by ELISA before and after treatment.

RESULTSChanges in blood flow of vertebral artery before and after treatment:there was no significant difference between model group and massage for 10 min group;there was significant difference between model group and massage for 20, 30 min groups(<0.01); there was significant difference between massage for 10 min group and massage for 20, 30 min groups(<0.01); there was no significant difference between massage for 20 min group and massage for 30 min group. Changes of NPY content before and after treatment: there was significant difference between model group and massage for 10 min group(<0.05); there was significant difference between model group and massage for 20, 30 min groups(<0.01); there was significant difference between massage for 10 min group and massage for 20, 30 min groups(<0.01); there was no significant difference between massage for 20 min group and massage for 30 min group. Changes of ET-1 content before and after treatment:there was no significant difference between model group and massage for 10 min group;there was significant difference between model group and massage for 20, 30 min groups(<0.01); there was significant difference between massage for 10 min group and massage for 20 min, 30 groups (<0.01); there was no significant difference between massage for 20 min group and massage for 30 min group.

CONCLUSIONSMassage needed 20 min for rabbits with CSA can only significantly improve the blood flow of vertebral artery. However, prolonging the time of massage has no obvious effect. The material basis of this timeliness characteristic of massage is closely related to the change of NPY and ET-1 levels in serum.

OBJECTIVETo investigate the role and molecular mechanism of membrane-associated guanylate kinase inverted 3 (MAGI3) in glioma cell proliferation.

METHODSThe expression levels of MAGI3 and PTEN were assessed in glioma samples by Western blotting. MAGI3 was stably transfected into C6 glioma cells to obtain C6-MAGI3 cells. Then, the proliferation, the expression levels of MAGI3 and PTEN, and Akt phosphorylation were evaluated in C6 and C6-MAGI3 cells. Xenograft tumor models were established by subcutaneous injection of C6 and C6-MAGI3 cells into nude mice, and the growth rates of xenografts in the mice were compared. The potential role of MAGI3 expression in PI3K/Akt signaling activation was further investigated by examining the correlation between MAGI3 expression and the expression of PI3K/Akt signaling downstream target genes in a glioma dataset using gene set enrichment analysis (GSEA).

RESULTSExpression levels of MAGI3 and PTEN were significantly downregulated in gliomas. Overexpression of MAGI3 in the glioma C6 cell line upregulated PTEN protein expression, inhibited the phosphorylation of Akt, and suppressed cell proliferation. MAGI3 overexpression also inhibited the growth of C6 glioma tumor xenografts in nude mice. Analysis based on the GEO database confirmed the negative correlation between activation of PI3K/Akt pathway and MAGI3 mRNA levels in human glioma samples.

CONCLUSIONThe loss of MAGI3 expression in glioma may enhance the proliferation of glioma cells via downregulation of PTEN expression, leading to the activation of the PI3K/Akt pathway. MAGI3 is a potential glioma suppressor.

Animals ; Brain Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; genetics ; Down-Regulation ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Membrane Proteins ; genetics ; metabolism ; Mice, Nude ; PTEN Phosphohydrolase ; genetics ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Signal Transduction ; Transfection ; Up-Regulation ; Xenograft Model Antitumor Assays

Objective To investigate the infection status of intestinal nematodes and their risk factors in rural residents of Jiangxia District, Wuhan City, so as to provide the reference for the establishment of targeted prevention and control measures. Methods According to the requirements of cross-sectional study of the national human key parasitic diseases, the residents in Shanpo Sub-district, Jiangxia District were randomly selected as the objects for the investigation of the infection status of Ascaris lumbricoides, hookworm and Trichuris trichiura. A questionnaire survey was performed simultaneously. Results Totally 236 residents received the stool tests, and only the hookworm eggs were found, with an infection rate of 7.63% (18 cases). The survey of intestinal parasitic infection status and their risk factors in 600 residents found 397 cases of suspected roundworm infection, 355 cases of suspected hookworm larvae dermatitis, 295 cases of suspected hookworm infection, and 145 cases of suspected whipworm infection, respectively. The Logistic regression analysis found that the risk factors of suspected roundworm infection included not always washing hands after toilet, using manure and fertilizer, and not often wearing shoes when doing farm work; the protective factors included knowing how roundworm infection was gained, and not using fresh dung for fertilization. The risk factors of suspected hookworm infection included using well water and pond water as the source of drinking water, eating food off the ground, using manure and fertilizers, and not washing when there was only one cutting board, and its protective factors included using tap water and well water as drinking water source, knowing how the roundworm infection was gained, and sometimes or not suing fresh feces for fertilization. The risk factors for suspected whipworm infection included eating food off the ground. Conclusions The intestinal parasite prevalence in the residents of the Shanpo Sub-district, Jiangxia District in Wuhan City is relatively high, and some risk factors associated with their infection indeed exist. Therefore, the regional government and health department should strengthen the health education to improve the residents’ knowledge about parasitic diseases and selfprotection awareness. At the same time, the residents should take physical examination regularly and take necessary deworming treatment to effectively control the prevalence and spread of intestinal parasitic diseases.