GJ-4 is crocin enrichments extracted from Gardenia jasminoides J. Ellis, and our previous studies have shown that GJ-4 significantly improved learning and memory impairment induced by Aβ in mice. Herein, a memory deficit model was developed by injecting okadaic acid (OA) into the lateral ventricle of mice, and the neuroprotection and underlying mechanism of GJ-4 on neuronal injury caused by Tau hyperphosphorylation were investigated. The Animal Care & Welfare Committee, Institute of Materia Medica, CAMS & PUMC has approved all procedures (No.00000318). GJ-4 at different doses was intragastric administration to mice for 16 days. Step-down test and Morris water maze test showed that GJ-4 could significantly improve OA-induced memory impairment in mice, and reduced the loss of Nissl bodies in the hippocampus of mice. GJ-4 could also decrease the phosphorylation level of Tau protein at Ser396, Thr231 and Ser404 via increasing protein phosphatase 2A (PP2A) activity and inhibiting glycogen synthase kinase-3β (GSK-3β) activity. Besides, further researches indicated that GJ-4 could inhibit the level of oxidative stress in the brain of OA mice, reduce neuronal apoptosis and inhibit the neuroinflammation mediated by activation of astrocytes in the hippocampus of mice, and eventually achieve its effects in improving learning and memory impairment in mice. According to these findings, we anticipated that GJ-4 might be a potential therapeutic drug for Alzheimer's disease.

BackgroundChoriodal neovascularization is an important ocular manifestation of angiogenesis in eyes,which derives from the choroid capillaries.Recent studies have found that complement activation is playing a key role in the laser-induced CNV.Because of the key position of CFB in the alternative pathway,bytargeting CFB and blocking the alternative pathway may provide an approach to observe the role of this alternative pathway in the generation of CNV.Objective This study was to investigate the inhibitory effect of reconstructed complement factor B (CFB)-small interfering ribonucleicacid(siRNA)on choroidal neovascularization (CNV)and its mechanism. Methods Experimental CNV was induced by laser photocoagulation in 96 eyes of 48 clean Brown Norway rats.The rats were randomly divided into 4 groups.25,50 and 75 μg B factor siRNA were injected via caudal vein on 1 day,3,5 days after photocoagulation in different dose groups,and normal saline solution was injected at the same way in experimental control group.Other 12 normal rats were used as blank control group.Fundus fluorescein angiography(FFA) was performed on 3,7,14,21,28 days after injection of CFB-siRNA and CNV was scored.The expressions of vascular endothelial growth factor(VEGF) and factor Ⅷ in choroid were detected by immunochemistry.The expressions of CFB-siRNA,VEGF,transforming growth factor β2( TGF-β2 )proteins in choroid were determined using immunochemistry in 7,14,21,28 days,and the expressions of mRNA of CFB-siRNA,VEGF,TGF-β2 were examined by reverse transcription polymerase chain reaction(RT-PCR). ResultsFFA revealed that the CNV rates in various doses of CFB-siRNA groups were significant lower than those of experimental control group in various time points(P＜0.05),and those in 75 μg B factor siRNA were decreased in comparison with 25 μg B factor siRNA (P＜0.05).Immunochemistry showed that the intensities of the VEGF and factor Ⅶ expression in various doses of CFB-siRNA groups were weaker than the blank control group ( P ＜ 0.05 ).Compared with the control group,the expression of CFB reduced in 7 days,and then approached to the level near the control group.Fourteen to twenty-one days after injection of CFB-siRNA,VEGF and TGF-β2 depressions in different doses of CFB-siRNA groups were lower than blank control group( P＜0.05 ).CFB expression in choroid showed the lower levels in CFB-siRNA injection group compared with blank control group in from 7 through 21 days (P＜0.05).RT-PCR displayed the gradual increase of CFB mRNA and curve-like changes of VEGF and TGF-β2 with time prolong. Conclusions Recombinated CFB-siRNA can effectively inhibit laser-induced CNV by down-regulating the expression of VEGF and factor Ⅷ.Alternative pathway of complement plays an important role in the production of CNV.

OBJECTIVETo describe the prevalence of anemia and yearly trends (1993-2003) among women who came to the hospitals or maternal and child health units for premarital examinations in 6 counties of Jiangsu and Zhejiang provinces.

METHODSData were obtained from the records of the premarital examinations in perinatal health care surveillance system that had been established since 1992 in these areas. We reviewed hemoglobin levels of those women who were enrolled in the perinatal health care surveillance system from 1993 to 2003. Anemia was defined according to the WHO (2001) criterion. We calculated the prevalence of anemia and analyzed the yearly trends based on the data of hemoglobin concentration.

RESULTSIn the period of 1993-2003, there were 82 995 anemia cases identified among 193,434 women with an overall anemia rate as 42.9%. The rates of anemia were high (65.5%) in 1993 but low (25.8%) in 2003. 99.7% of the anemic women whose hemoglobin concentration were between 80-119.9 g/L. Time trend analysis indicated a significant decline on anemia rate while monthly analysis showed that the prevalence rates were high (48.2%) in September and low in March (39.5%). The results also showed that the prevalence rates of anemia were relatively higher in farmers and workers in rural enterprises, and lower in Han ethnicity than minorities. The higher prevalence rates of anemia were presented among the women with less education, lower body mass index, or at older age.

CONCLUSIONFor those premarital women in 6 counties of Jiangsu and Zhejiang provinces, the overall anemic rate presented a significant downward trend between 1993 and 2003 while the prevalence of anemia remained high, especially for the women with less education, lower body mass index or older ages.

Adolescent ; Adult ; Anemia ; epidemiology ; China ; epidemiology ; Female ; Humans ; Middle Aged ; Premarital Examinations ; Young Adult

OBJECTIVETo analyze the early clinical effects of total hip arthroplasty(THA) for the treatment of old acetabular fractures.

METHODSFrom January 2007 to June 2010, thirteen patients with old acetabular fractures were reviewed, including 10 males and 3 females. Ten patients were treated with internal fixation and conservative treatment had been used in three patients. The average Harris Hip Score was used to evaluate therapeutic effects.

RESULTSAfter operation, all thirteen patients were followed up for one year. Hip X-ray films were taken and prosthesis loosening was not seen on any of the films at the 1st year after operation. The Harris Hip Score improved from preoperative (37.19 +/- 20.12) to postoperative (83.38 +/- 3.33), there was statistically significant difference.

CONCLUSIONFor reasons of malunion or failure of internal fixation, large and various bone defect, it's difficult to reach the anatomical reduction. THA is a good treatment method, but it needs rich skills and experience compared with ordinary operation.

Acetabulum ; injuries ; Arthroplasty, Replacement, Hip ; methods ; Female ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Retrospective Studies

Objective To observe the T cells immune response to Enolase (Eno),an immunodominant antigen of Candida albicans.Methods Determined the frequencies of positive spot-forming cells (SFCs) of Eno antigen-specific T cells secreting IFN-γ,IL-4 and IL-17A in the PBMCs of 25 healthy individuals by ELISPOT assay.Results After Eno stimulation,the SFCs of IFN-γ,IL-4 and IL 17A in 25 healthy persons were 14.00(8.50,39.00),0(0,0) and 2(1,4.50),respectively.Either the SFCs of IFN-γ or those of IL-17A were significantly higher than those of IL-4 (P＜0.05).The difference between SFCs of IFN-γ and those of IL-17A was also significant (P=0).The response rates of IFN-γ,IL-4 and IL-17A were 100％ (25/25),4.00％ (1/25) and 88.00％ (22/25),respectively.The difference between either IFN-γor IL 17A and IL-4 was significant (values all P＜0.05).Eno induced strong response (SCFs≥20) for IFN-γ in 10 healthy individuals (40.00％,10/25),but failed to induce strong response for IL-17A and IL-4 in all the volunteers.Major healthy individuals (84.00％,21/25)showed both Th1 and Th17 cells response against Eno,12.00％ (3/25) showing Th1 cells response in isolation,and none showed Th2 or Th17 cells response individually.Conclusion Eno of Candida albicans could induce immunodominant responses of Th1 and Th17 cells,which was considered to provide protection to IC.Eno might be a potential protective vaccine against IC.

Objective To observe the effects of high fat dietinduced elevation of blood glucose on the microvascular function of testis and male reproduction in C57BL/6 mice. Methods A total of 40 male C57BL/6 mice were randomly divided into control group and high fat diet (HFD) group (n =20). The mice in HFD group were fed with high fat diet for 20 weeks. Blood glucose and body weight were measured weekly. The permeability of bloodtestis barrier was evaluated by intraperitoneal injection of Evans blue. The blood flow of testicular microcircu-lation and the frequency and amplitude of microvascular vasomotion were detected by laser Doppler blood flow ima-ging system. The morphology of testicular tissue was observed by HE staining. The expressions of platelet- endothelial cell adhesion molecule-1 (PECAM-1/CD31) in testicular microvascular endothelial cells and prolifera-ting cell nuclear antigen (PCNA) in spermatogenic cells were detected by immunohistochemistry. The apoptosis of spermatogenic cells was observed by TUNEL staining. Results The body weight and blood glucose of HFD group were significantly higher than those in control group (P＜0.01). Evans blue staining showed that the integrity of blood-testis barrier of HFD group was damaged, and increased permeability was observed in seminiferous tubules. In HFD group, the mean blood flow of testis and the frequency and amplitude of microvascular vasomotion were sig-nificantly lower than those in control group (P＜0.01). The number of spermatogenic epithelial cells and the thickness of seminiferous epithelium decreased. The expressions of CD31 in microvascular endothelial cells and PCNA in spermatogenic cells were significantly lower in HFD group than those in control group (P＜0.01). The apoptosis level of spermatogenic cells was higher than that in the control group (P <0.01). Conclusions Increased blood glucose level induced by high fat diet in mice can impair the testicular microvasculature and damage the integrity of blood-testis barrier and injure the structure of seminiferous epithelium in mice.

In order to make Dendrobium officinale return to the nature, the temperature and humidity in whole days of the built rock model with different slopes and aspects in the natural distribution of wild D. officinale in Tianmu Mountain were recorded by MH-WS01 automatic recorder. The results showed that the slope has a significant impact on the extreme temperature on the surface of the rocks. In summer, the extreme temperature on the surface of horizontal or soft rock can reach to 69.4 ℃, while the temperatures were lower than 50 ℃ on the vertical rock. In winter, the temperatures on the surface of vertical rock were higher and the low temperature duration was shorter than those on the horizontal or soft rock. Also, the humidity of the rocks was significantly influenced by the slope. The monthly average humidity on the surface of vertical rock was above 80%RH. Furthermore, the aspect had a significant impact on the temperature and humidity on the surface of the rocks, but had no significant effect on the daily mean temperature and extreme temperature on the surface of vertical rock. Therefore, the slope affects the survival of D. officinale by affecting the extreme temperature of rocks and affects the growth of D. officinale by affecting the humidity. The choice of slope is the key to the success of cliff epiphytic cultivation for D. officinale.

OBJECTIVES: Ziyin Huatan Recipe (ZYHT), a traditional Chinese medicine comprised of Lilii Bulbus, Pinelliae Rhizoma, and Hedyotis Diffusa, has shown promise in treating gastric cancer (GC). However, its potential mechanism has not yet been clearly addressed. This study aimed to predict targets and molecular mechanisms of ZYHT in treating GC by network pharmacology analysis and to explore the role of ZYHT in GC both in vitro and in vivo. METHODS: Targets and molecular mechanisms of ZYHT were predicted via network pharmacology analysis. The effects of ZYHT on the expression of metastasis-associated targets were further validated by Western blot and quantitative real-time polymerase chain reaction. To explore the specific molecular mechanisms of the effects of ZYHT on migration and invasion, the runt-related transcription factor 3 (RUNX3) gene was knocked out by clustered regularly interspaced short palindromic repeats/Cas9, and lentiviral vectors were transfected into SGC-7901 cells. Then lung metastasis model of GC in nude mice was established to explore the anti-metastasis effect of ZYHT. Western blot and immunohistochemistry were used to explore the impact of ZYHT on the expression of metastasis-related proteins with or without RUNX3 gene. RESULTS: The network pharmacology analysis showed that ZYHT might inhibit focal adhesion, migration, invasion and metastasis of GC. ZYHT inhibited the proliferation, migration and invasion of GC cells in vitro via regulating the expression of metastasis-associated targets. Knocking out RUNX3 almost completely reversed the cell phenotypes (migration and invasion) and protein expression levels elicited by ZYHT. In vivo studies showed that ZYHT inhibited the metastasis of GC cells to the lung and prolonged the survival time of the nude mice. Knocking out RUNX3 partly reversed the metastasis of GC cells to the lung and the protein expression levels elicited by ZYHT. CONCLUSION ZYHT can effectively inhibit the invasion and migration of GC in vitro and in vivo, and its molecular mechanism may relate to the upregulation of RUNX3 expression.
Animals ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; China ; Gene Expression Regulation, Neoplastic ; Mice ; Mice, Nude ; Neoplasm Invasiveness ; Stomach Neoplasms/genetics*

To establish a quantitative analysis of multi-components by single marker(QAMS) method for five flavonoids in Rhododendron anthopogonoides and verify its feasibility and applicability in the medicinal materials of R. anthopogonoides. With hyperoside as the internal reference, relative correction factors(RCF) of rutin, quercetin, quercitrin and kaempferol were established by high-performance liquid chromatography(HPLC) analysis. RCFs were used to calculate the content of each component, system durability and relative retention time. Simultaneously, QAMS and external standard method(ESM) were used to determine the content of five flavonoids in 12 batches of R. anthopogonoides from different origins. The results were statistically analyzed to verify the accuracy and feasibility. The fingerprints and cluster analysis data of R. anthopogonoides analyzed and discussed differences among the batches. According to the results, the RCFs of rutin, quercetin, quercetin and kaempferol in R. anthopogonoides were 1.242 6, 0.990 5, 0.535 0, and 0.781 3, respectively. The RCFs represented a good reproducibility under different experimental conditions. Besides, there was no significant difference between QAMS and ESM. Besides, the fingerprint and cluster analysis data showed the consistency between the classification and with the origin distribution of the herbs. In conclusion, the QAMS method shows a good stability and accuracy in the quality control of R. anthopogonoides.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Flavonoids ; Medicine, Tibetan Traditional ; Reproducibility of Results ; Rhododendron

MCT is an important key enzyme in the terpenoid biosynthesis in MEP pathway. In this study, Gateway technology was used to construct RNAi vector of TwMCT, and a vector fragment with a size of 484 bp was obtained. The TwMCT RNAi vector was transferred into the suspension cells of Tripterygium wilfordii by gene gun. Accumulation of terpenoids was assayed by UPLC, and the result showed that the content of triptolide and celastrol in cells decreased by 23.4% and 42.8%, respectively, compared with the control group pK7GWIWG2D. Moreover, the gene expression of TwMCT and major genes in terpenoid biosynthesis pathway was detected by qRT-PCR, which demonstrated that the expression of TwMCT reduced by 29.2% relative to that of the control group pK7GWIWG2D, and the relative expression of TwDXR, TwGGPS, TwHMGR and TwHMGS diminished by 36.3%, 31.3%, 62.2%, and 29.1%, respectively, but the expression of TwDXS was up-regulated by 114.2%, and there was no significant change in TwFPS. Thus, it was verified in vivo that interference with TwMCT expression significantly inhibited the accumulation of triptolide and celastrol in Tripterygium wilfordii, laying a foundation for further exploring the regulation mechanism of MCT gene on the terpenoid biosynthesis in Tripterygium wilfordii.