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We synthesized eight tanshinone anhydrides and the alcoholytic derivatives through a mild oxygen-insertion under Pd/C catalytic hydrogenation conditions. The suppressive effects of the anhydrides on the oxidized low-density lipoprotein (oxLDL) uptake and the oxLDL-induced macrophage-derived foam cell formation were studied. Our results revealed that both anhydrides 1a and 2a could significantly suppress the oxLDL uptake in macrophages and the foam cell formation at micromolar level, which might be partially attributed to their inhibition of oxLDL-induced LOX-1 expression in macrophages.

In the title complex, [Cu(C22H17Br2N2O2)2], the Cu(II) ion is four-coordinated in a trans-CuN2O2 square-planar geometry by two phenolate O and two imino N atoms from two deprotonated N,O-bidentate ligands. In the crystal, the packing of the mol-ecules is controlled by C-H⋯π and π-π inter-actions [centroid-centroid distances = 3.568 (3), 3.678 (2), 3.717 (3) and 3.799 (2) Å] and weak Br⋯Br halogen bonds [3.508 (4) Å], linking the mol-ecules into an infinite three-dimensional supra-molecular network.

Objective: To analyze the factors affecting pathologic complete response (pCR) to neoadjuvant chemotherapy (NAC) in breast cancer patients., Methods: A retrospective cohort study was carried out to analyze the clinical data of 141 breast cancer patients treated with neoadjuvant chemotherapy. The factors affecting pCR and the changes of tumor receptor status before and after treatment were analyzed., Results: Among all the 141 patients, 21 patients (14.9%) achieved pCR. The rate of pCR achieved by regimens of anthracycline combined with taxane was higher (16.8%, 19/113) than that by anthracycline-containing regimens (7.1%, 1/14). The dose intensity of anthracycline had a significant correlation with pCR rate (P < 0.05). The pCR rate in the relative dose intensity of taxane ≥ 0.85 arm was higher than that of < 0.85 arm (P = 0.02). Eighty patients (56.7%) had completed more than 4 cycles of chemotherapy and the median time to achieve pCR was 6 (3 to 10) cycles. The pCR rate had a significant difference between patients < 6 and ≥ 6 cycles (7.1% vs. 22.5%,P = 0.01). Multivariate analysis showed that tumor size measured by palpation ≤ 5 cm and ≥ 6 chemotherapy cycles were significantly related with pCR rate (P < 0.05). In all the 21 pCR patients, the pre-treatment ER(-), PR(-), HER-2(-) statuses were in 14, 14 and 17 patients, respectively. The status of ER, PR, HER-2 of most patients (74.2%, 69.7% and 87.7%, respectively) was not changed after treatment. Among the patients with changes in receptor status, ER changed from negative to positive was in the majority (37.1%, 13/35 vs. 12.9%, 4/31, P < 0.05), and the percentage of changes in PR and HER-2 status had no significant differences., Conclusions: The regimens of anthracycline combined with taxane can achieve a higher pCR rate. The lymph node and receptor status before therapy have no significant correlation with pCR. Patients who have primary tumor size ≤ 5 cm, ≥ 6 chemotherapy cycles and enough dose intensity are easier to achieve pCR. The receptor status before and after therapy should be determined, and according to any positive results, physicians can chose HER-2 targeted therapy and/or endocrine therapy after surgery to benefit the patients.

To construct the adenoviral vector with co-expressing keratinocyte growth factor (KGF) and enhanced green fluorescent protein (EGFP) for transfection into the mesenchymal stem cells (MSC), the target gene KGF was cloned into the shuttle plasmid with the report gene EGFP, then the recombinant shuttle plasmid was transformed into DH5a bacteria to recombine with backbone vector pAdxsi. Next, the plasmid pAd-EGFP-mKGF was amplified in H293 cells and the viral titer was determined. The MSC were separated and enriched by using bone marrow adherent culture and identified in vitro to observe the efficiency of transfection. The results indicated that the recombinant shuttle plasmid pShuttle-EGFP-mKGF digested with restriction endonucleases was confirmed by two products which length was about 0.6 kb and 5.1 kb, respectively; the recombinant plasmid pAdxsi-EGFP-mKGF digested with restriction endonucleases was confirmed by 7 products; recombinant adenoviral vector Ad-EGFP-mKGF was amplified to titer of 1.6 × 10(10) pfu/ml. At 10 h after transfecting MSC began to express fluorescence at 6 to 8 days later, the fluorescence reached to the peak with infection rate of 92.3, at 28 days the expression of fluorescence was still observed. It is concluded that the recombinant adenoviral vector Ad-EGFP-mKGF is successfully constructed and can transfect MSC effectively and safely.

Primary hepatocytes are commonly used during in vitro studies, but care must be taken with isolation and culture of the cells to ensure their viability. In this study, hepatocytes were isolated from the liver (caudate process) of a newborn calf by the collagenase perfusion and digestion method. The trypan blue exclusion method was used to determine total cell number and the survival rate of hepatocytes, while hepatocyte function was assessed by measuring lactate dehydrogenase, albumin and urea in culture medium supernatants at 24, 48, 72, 96, 120, 144 and 168 h. Results showed that the number of viable cells/g of liver (wet weight) averaged 1.12×10(7) cells/g, with an average hepatocyte viability of 85.7% (range 83-92%). After 48 h of culture, the hepatocytes solidly adhered to the well culture plate and were spread in an epithelioid shape, with clear cell boundaries between the cells and biliary ductule-like structures formed which persisted for up to 10 days. Hepatocyte function was optimal at 72 h after isolation and culture. This simple and economical procedure for the isolation and culture of viable cells may be useful for in vitro bovine hepatocyte studies., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Objective: To explore the feasibility of adipose-derived stem cells (ASC) combined with micronized acellular dermal matrix (MADM) for vocal cord injection., Methods: The adipose-deprived stem cells were harvested from rabbit adipose tissue in vitro. The 3rd generation of ASC was labeled with DiI (1,1-dioctadecyl-3,3,3,3-tetramethylindocarbocyanine perchlorate) and cultured with MADM to form a complex. The adhesion of ASC to MADM was observed by fluorescence microscope and electron microscope. The proliferation of ASC on MADM was evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy methoxyphenyl)-2-(4-sulfonyl)-2H-tetrazolium, inner salt (MTS). Three days after the culture, the complex was mixed with appropriate amount of collagen, and then injected into the unilateral vocal cord of the rabbit. The animals were sacrificed 2, 4, 8 weeks after injection, the survival time and distribution of ASC in vocal fold were tested, and the responses of vocal cord to ASC-MADM and the degradation of MADM were observed., Results: The ASC adhered to MADM and grew well (P < 0.05 or < 0.01), showing good compatibility with MADM in vocal cord tissue. The complex of ASC-MADM could be injected into the rabbit vocal cords, while no adverse reactions was observed in the vocal cord by endoscope, frozen section and HE staining. ASC could survive for 8 weeks in vocal cords, and no inflammatory cell infiltration was observed., Conclusions: MADM is an ideal scaffold material and shows perfect compatibility with ASC which can adhere and proliferate well on it. The complex of ASC-MADM can be injected into the vocal cord and can survive. There is no adverse reaction in vocal cords.

Objective: To observe the effects of Radix Glycyrrhizae polysaccharide on regulatory T cells (Treg) in spleen and lymphocyte transformation ratio in tumor-bearing mice so as to explore the mechanisms of its immunoregulatory function., Methods: Fifty BALB/c mice were randomly divided into normal group, untreated group, cyclophosphamide group, Radix Glycyrrhizae polysaccharide group and Radix Glycyrrhizae polysaccharide plus cyclophosphamide group. Except normal group, mice were subcutaneously implanted H22 tumor cells in the right axillary region. After 24 h, mice in normal and untreated group were subcutaneously injected with physiological saline, while mice in the cyclophosphamide group were intraperitoneally injected with cyclophosphamide and mice in Radix Glycyrrhizae polysaccharide group were subcutaneously injected with polysaccharide. Fourteen days later, Treg cells of spleen were detected by flow cytometry and lymphocyte transformation ratio was detected by methyl thiazolyl tetrazolium method., Results: The proportion of Treg cells was significantly higher in the untreated group than in the normal group, and was lower in the Radix Glycyrrhizae polysaccharide group than in the untreated group (P < p0.01). Lymphocyte transformation ratio in the Radix Glycyrrhizae polysaccharide group was higher than that in the cyclophosphamide group. There was no interaction between Radix Glycyrrhizae polysaccharide and cyclophosphamide., Conclusion: Radix Glycyrrhizae polysaccharide can regulate the cellular immunity disorders of tumor-bearing mice by decreasing proportion of Treg cells and increasing spleen lymphocyte transformation ratio.

Objective: To compare the efficiency of response evaluation by clinical examination, ultrasonograghy and mammography in neoadjuvant chemotherapy (NAC) for breast cancer., Methods: A retrospective cohort study was conducted to analyze the data of 141 patients treated with neoadjuvant chemotherapy. Response evaluation was performed by clinical palpation, ultrasound and mammography., Results: Only 12 (8.5%) among the 141 patients presented with a stage I tumor. The tumor size determined by palpation was often larger than that by ultrasound before therapy (P < 0.01). Among patients with suspicions axillary nodes checked by ultrasound, 88.3% (53/60) of them had positive nodes by pathology before NAC, and 34.5% (10/29) of patients with negative nodes determined by ultrasound had positive nodes by pathology. In all the 141 patients, 21(14.9%) showed pathological complete remission in both the primary tumor and lymph node. For response evaluation, the false complete remission rate judged by clinical examination was 46.8% (22/47), and the false tumor residual rate by ultrasound was 84.0% (21/25). In 53.5% (23/43) of patients the response could not be assessed by mammography due to that the tumors were undistinguishable in size. The range of microcalcification was not reduced in 5 patients with a partial response of the tumor. 25 patients experienced needle puncture during therapy. Among them, in the 9 pathologically negative patients, only 3 achieved pCR, and the other 16 positive patients didn't achieve pCR., Conclusion: Using the puncture or sentinel lymph node biopsy, clinicians should pay enough emphasis on the pathological determination of the node status before chemotherapy. Clinicians will make a quite of false judgment of the tumor by clinical examination, ultrasound or mammography. They may use needle puncture during therapy to evaluate the response of neoadjuvant chemotherapy, and the result should be analyzed synthetically.

Objective: To evaluate the roles of enteric nervous system neurotransmitters, nitric oxide (NO), substance P (SP) and vasoactive intestinal polypeptide (VIP), and interstitial cells of Cajal (ICC) in the colon in slow transit constipation in rats., Methods: Thirty-two healthy Wistar rats were randomly assigned to control and constipated groups. In the constipated group, the rats were daily administered with diphenoxylate (8 mg/kg) to develop slow transit constipation, while the control rats were fed with water. The number and the weight of fecal granule and the body weight of rats were recorded every 5 days for 90 days. Transit functions of intestinal movement were examined by an activated charcoal suspension pushing test one week after stopping the administration of diphenoxylate. The levels of NO and SP in the colonic mucosa were measured by nitrate reductase methods and ELISA respectively. The distribution of VIP and ICC positive cells confirmed with symbolic c-kit+ cells in the colonic wall were observed by immunohistochemical methods., Results: The daily number of fecal granule in the constipated group was significantly less than that in the control group (P<0.01). The mean weight of each fecal granule in the constipated group was significantly higher than that in the control group (P<0.01). The discharge time of the first granule of black faeces in the constipated group (430.2+/- 132.1 min) was significantly longer than that in the control group (337.2+/- 74.7 min; P<0.05). There were no significant differences in NO and SP levels and the density of VIP positive cells in the distal colonic segment between the two groups. The number of c-kit+ cells in the distal colonic wall in the constipated group was significantly reduced compared with that in the control group (P<0.05)., Conclusions: The reduction of ICC number in the distal colon may be contributed to the pathogenesis of slow transit constipation in rats.

Objective: To study the association of vertical facial skeletal types and sagittal facial skeletal types with anterior alveolar bone thickness., Methods: Among 168 cases with malocclusion in early permanent dentition stage, 93 patients were male and 75 patients were female. All patients (aged 10-14 years) were divided into 9 groups by different facial skeletal types, mandibular anterior alveolar bone thickness in patients' lateral cephalometric films were measured. ANOVA were performed to measurement results with the SPSS 13.0 statistical software., Results: Though sagittal facial skeletal types were the same, there were significant differences between different vertical facial types groups. The order was low-angle group, average-angle group and high-angle group according to the size. A high-angle individual often had a thin anterior alveolar bone while a low-angle individual often had the opposite morphology character. There was no statistical significance between skeletal type I, II and III. But group of skeletal type III also had a thin alveolar bone thickness which had no significant difference with high-angle group. Low-angle group III had no significant difference with average-angle group I and II in alveolar bone thickness., Conclusion: Sagittal facial skeletal types have little influence on anterior alveolar morphology, but the vertical facial skeletal types have strong connection with anterior alveolar bone thickness.

By skin prick test, three kinds of mite allergens (Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicalis) were tested in a group of asthma children in Jinhua area from Oct 2005 to Sep 2006. The positive rate to allergen from D. pteronyssinus and D. farinae was 80.6% and 77.8% respectively, higher than that of Blomia tropicalis (61.1%) (chi2 = 21.39, P < 0.05). Cases positive to Blomia tropicalis allergen showed 100% and 95.9% positive reaction to D. pteronyssinus and D. farinae respectively. The results demonstrated that the important allergens for children's asthma are from D. pteronyssinus and D. farinae, while the with the former two mite allergens. Blomia tropicalis allergen is supposed to have cross-reactivity

By the methods of remote sensing (RS) and geographic information system (GIS), and based on the estimations of degradation degree, risk degree and easy-restoration degree of degraded grasslands, an ecological management index (EMI) model of grassland was established to approach the practical ways of optimizing management of degraded grassland. A case study in the Xilin River Basin of Inner Mongolia showed that this model could quantitatively analyze the degradation degree, risk degree and easy-restoration degree of the grasslands under different optimizing management levels, which was of significance for applying rational measures with pertinence, and beneficial to the optimal allocation of resources during the management of degraded grassland. The EMI model could integrate most concerned information, which made it applicable widely.

Objective: Through comparison of HER2/neu oncogene detected by chromogenic in situ hybridization (CISH) and immunohistochemistry (IHC) in breast cancer, to explore the effect of CISH on detecting gene amplification of HER2., Methods: Selected formalin-fixed paraffin-embedded breast samples whose pathological types were infiltrating ductal carcinomas (255 retrospective samples, 271 prospective samples), and these samples were detected by IHC and CISH., Results: (1) In the retrospective study, CISH identified gene amplification in 91.6% of IHC score 3+ tumors (120/131) and in 56.5% of IHC score 2+ tumors (39/69), thus the concordant ratio between IHC and CISH was 81.2% (207/255). The two results showed significant correlation (P<0.01). (2) In the prospective study, the ratio of HER2 protein over expression detected by IHC was 31.7%, the ratio of HER2 gene amplification detected by CISH was 27.3%. CISH identified gene amplification in 91.4% of IHC score 3+ tumors (53/58) and in 46.4% of IHC score 2+ tumors (13/28), Concordant ratio between IHC and CISH was 89.7% (243/271). Two results showed significant correlation (P<0.01). (3) Paired CISH/FISH results were concordant in 14 of 15 cases. The remaining case was detected by FISH, but showed no HER2 gene amplification by CISH. (4) The gene amplification by CISH had a significantly reverse correlation with ER and PR expression (P<0.01)., Conclusions: The results of HER2 gene amplification detected by CISH have high concordance with the results detectd by IHC and FISH. CISH is a novel technique for detecting HER2 gene amplification.

Objective: To investigate the expression of oncoprotein c-erbB2 in primary breast cancer and to analyze its relation to its prognosis., Methods: Immunohistochemical staining for c-erbB2 was performed on paraffin-embedded specimens of primary breast cancer from 284 patients, and the relation to its prognosis was statistically analyzed., Results: Positive expression rate of c-erbB2 was 26.8% (76/284) in 284 primary breast cancer patients. Expression of c-erbB2 was positively correlated with the status of lymph node metastasis (P = 0.003). Univariate analysis indicated that c-erbB2 expression is a significant prognostic factor for the disease-free survival (DFS) (P = 0.024) and overall survival (OS) (P = 0.002), while multivariate analysis demonstrated that c-erbB2 is an independent prognostic factor for OS (P = 0.023). Moreover, tumors with c-erbB2 positive expression are more tend to metastasis to other viscera than those with c-erbB2 negative. c-erbB2 expression has different prognostic values for patients with different status of estrogen receptor (ER) and lymph node metastasis., Conclusion: c-erbB2 expression is an independent prognostic factor for total survival time in primary breast cancer patients, and its prognostic values are different according to the different ER status and lymph node metastasis.

The dominant gene Xa21 with broad-spectrum and high resistance to Xanthomonas oryzae pv. oryzae (Xoo) was transferred into C418, an important restorer line of japonica hybrid rice in China using double right-border (DRB) T-DNA binary vector through Agrobacterium-mediated transformation. 17 transgenic lines were Xa21-positive with high resistance to the race P6 of Xoo through PCR analysis and resistance identification, among the total 27 independent primary transformants (T0) obtained. The subsequent analysis of the T1 progenies of these 17 T0 lines through PCR-assisted selection and resistance investigation showed that four Xa21 transgenic T0 lines could produce selectable marker-free (SMF) progenies. The frequency of primary transformants producing SMF progenies was 15%. In addition, PCR analysis also revealed these SMF progenies did not contain vector backbone sequence, and they were named as SMF and vector backbone sequence-free (SMF-VBSF) Xa21 transgenic plants. The further molecular and phenotypic analysis of the T2 and T3 progenies testified the homozygous SMF-VBSF Xa21 transgenic plants were obtained with high resistance to Xoo.

Objective: To discuss the skeletal and dentoalveolar characteristics of asymmetric mandibular excess malocclusions and to discuss the procedures of combined orthodontic-orthonganthic surgery treatments of asymmetric mandibular excess malocclusions., Methods: 25 cases treated by combined orthodontic-orthognathic surgery treatments were reviewed to find out the specialties of this kind of therapy., Results: The asymmetric of mandible presents anterior and posterior teeth tipped both sagitally and horizontally, as well as upper and lower jaws incompatibility. The pre-surgical orthodontic treatments included decomposition of anterior and posterior teeth, leveling and aligning the teeth etc. The post-surgical orthodontic treatments were to detail the occlusions. The patients all got functional and aesthetic good results after the combined orthodontic-orthognathic surgery treatments., Conclusion: The asymmetric mandibular excess affects the harmony of the face badly, and the correction of it must be carried out by the combined orthodontic-orthognathic surgery treatments. The pre- and post-surgical orthodontic treatments are the key stages to make the skeletal corrections stable.

Objective: To investigate the therapeutic effects of a nerve growth factor (NGF) isolated and purified from the venom of Naja naja atra on injured sciatic nerves in adult cat., Methods: Model of sciatic nerve crush lesion in 20 cats was made. After the operation, in the therapeutic group, NGF(2 micrograms/kg) was injected intramuscularly into the cat's leg once a day for 10 d (n = 5) and for 30 d (n = 5) respectively; in the control group, NGF was not injected and the cats were allowed to survive 10 d (n = 5) and 30 d (n = 5) respectively., Results: Ten days after the operation, the number of distal nerve fibers was significantly smaller in the control group than in the therapeutic group (P < 0.01); the reaction to planta stimulation appeared earlier and the leg action recovered faster in the therapeutic group. Thirty day after the operation, the number of distal nerve fibers in the therapeutic group was significantly greater than that of other groups (P < 0.01), but the histologic structure of the nerve fibers was in disorder and the axon and Ranvier node disappeared. About 16 days after injury and injection of NGF for consecutive days, the reaction to planta stimulation disappeared and leg paraplegia occurred in the operated side., Conclusion: The results showed that NGF could obviously reduce the degeneration of nerve fibers and enhance the peripheral nerve regeneration and functional recovery from ingury early, but daily injection of NGF in the injured region for a long period could significantly result in over-regeneration of nerve fibers, and the conductive function of the injured peripheral nerves would be lost.

The bio-safety of selective markers in transgenic plants has been a hot spot in the field of plant genetic engineering. To solve the problem of selective markers in the transgenic plants, two means of producing transgenic plants have been developed. One is the utilization of bio-safe positive selective markers which are genes mainly related to metabolism of auxins and carbohydrates. The other is the establishment of transformation systems allowing marker genes to be eliminated from the transgenic plants, which include co-ransformation, double T-DNA border vectors, site-specific recombination and transposition. All these approaches of plant genetic engineering will benefit breeding transgenic plants with bio-safety.

The transformation mediated by Agrobacterium has been successfully applied to rice in recent years. In the previous research we have transferred the Xa21 gene into five rice varieties of China, using Agrobacterium-mediated trasformation. In this study, T-DNA flanking sequences of Xa21 transgenic rice lines were obtained by using thermal asymmetric interlaced PCR (TAIL-PCR). The flanking sequences which are actual rice DNA were identified and located on molecular linkage map developed from a ZYQ8/JX17 double haploid (DH) population. A total of 22 T-DNA flanking rice sequences were isolated. Nineteen of them displayed RFLPs between the two parents, ZYQ8 and JX17, and were mapped on the rice chromosomes, 3, 4, 7, 9, 10, 11 and 12, respectively. The genetic mapping of T-DNA integration sites in Xa21 transgenic rice will benefit the study of position effect and stable inheritance of the transgene Xa21.

Flax (Linum usitatissimum L.) is an important fiber and oil-producing crop. Flax rust, caused by Melampsora lini Ehrenb. Lev., occurs worldwide and can cause severe losses in seed yield and fiber quality. In order to identify molecular markers linked to the flax rust resistant gene M4, RAPD analysis of NM4, a near-isogenic line containing the M4 gene, and the recurrent parent Bison was carried out with 540 decamer primers. The primer OPA18 could stably amplify a specific fragment, OPA18(432), in the NM4 line. The OPA18(432) marker was testified to be closely linked to the M4 gene with a genetic distance of 2.1 cM through the analysis of the F2 mapping population derived from a cross of Bison x NM4. Based on the sequence of OPA18(432), the specific PCR primers were designed, and a SCAR marker for the M4 gene was produced. Amplification of different resistant materials proved that the maker is specific for the M4 gene. This marker has been used successfully in marker-assisted selection in the flax breeding program.