We have developed a sensitive and rapid lateral-flow immunoassay (LFIA) for WSSV,using colloidal gold as an indicator.The fusion protein,VP (19+28),was expressed in E.coli,purified and used to prepare polyclonal antibodies.The purified anti-VP (19+28) IgG were conjugated with colloidal gold.Unconjugated anti-VP (19+28) IgG and goat anti-rabbit IgG were immobilized on nitrocellulose membranes.After assembly,three groups (5 individual animals in each group) of shrimp samples were tested which included healthy,moribund and dead shrimps.For each group,three different tissues (body juices,gills and hepatopancreas) were tested at the same time.In parallel,all the samples were also analyzed using PCR for comparison.Out of 45 samples tested,30 were detected as positive while 15 were classified as negative.The results of LFIA correlate with those obtained by the PCR analysis,indicating that these two detection methods have the same efficacy in the limited number of samples tested in this preliminary study.

Objective:To analyze risk factors of breast ductal carcinoma in situ (DCIS) with microinvasion (DCIS-MI) and explore suitable axillary lymph node surgery treatment for patients with DCIS-MI. Methods:The clinical characteristics, such as age, menopausal status at diagnosis, size of breast mass, and pathology reports of 45 patients with breast DCIS or DCIS-MI treated at Jinling Hospital, Medical School of Nanjing University from February 2013 to February 2016, were retrospectively collected and analyzed statistically to deter-mine the risk factors associated with microinvasion. Results:Premenopause (P=0.006), tumor size≥3.15 cm (P=0.006), and family his-tory of malignant tumor (P=0.002) were proven risk factors of DCIS-MI. Conclusion:Patients with clinical palpable axillary mass, pre-menopause, large breast mass, and family history of malignant tumor demonstrated high possibility of DCIS-MI. Hence, sentinel lymph node biopsy should be performed. Axillary lymph node dissection is highly recommended to patients whose main symptom is palpable axillary mass.

Objective To investigate the effects of dizocilpine(MK801) on depressive-like behaviors and damaged hippocampus in rats with diabetes-related depression.MethodsThe animal models of diabetes-related depression were established and they were randomly divided into two groups based on random number table: model group and MK801 group,while 6 rats were included in each group.And another six health rats were regarded as control group.The Open-field test was used to detect the activities.The damage of hippocampus was valued by HE staining,Nissl staining,and Tunel staining.The protein expressions of Bax,Bcl-2 in hippocampus were detected by Western blot.ResultsThe number of activities was significant decreased in Open-field test in model group when compared with control.Hippocampal neurons vacuoles,Nissl bodies were decreased and apoptotic cells were increased in hippocampus in model group as well.Furthermore,the expression of Bax was significant up-regulated(94.57±7.97,P<0.01),while the Bcl-2 was declined(24.65±5.26,P<0.01).Compared with the model group,the animals in MK801 group exhibited increased activities(12.50±4.42,P<0.01),which accompany with an increased Nissl body(133.55±16.74,P<0.01) and a decreased apoptosis(22.50±6.35,P<0.01).Moreover,the expression of Bax was decreased and the Bcl-2 was increased in MK801 group when they were contrasted to model(33.00±4.57,P<0.01).Conclusion MK801 is a significant element to regulate the expression of apoptosis protein including Bax,Bcl-2,and to protecte the hippocampal neuron in rats with diabetes-related depression effectively.

Objective To observe the effects of Baishile Capsules on the hippocampal PI3K signaling pathway in chronic mild unpredictable stress (CUMS) rats; To discuss its mechanism of action for anti-depression. Methods SD rats were randomly divided into 6 groups, control group, model group, fluoxetine group, Baishile Capsules high-, medium- and low-dose groups, with 10 rats in each group. The cums depression model was established, and gavage for medication was conducted at the same time for 21 days in a row. The behavioral changes of rats in each group were detected by the novel feeding experiment and Open-field experiment; monoamine neurotransmitter in serum were detected by ELISA; the expressions of PI3K, AKT and SGK1 in rat hippocampus were detected by immunofluorescence and Western blot test. Results Compared with the control group, feeding latency of rats in model group was significantly prolonged (P<0.01), horizontal and vertical activities were significantly reduced (P<0.01); the content of serum monoamine neurotransmitter decreased (P<0.01); the expressions of PI3K and AKT in hippocampus decreased, while the expression of SGK1 increased (P<0.01, P<0.05). Compared with the model group, Baishile Capsule high- and medium-dose groups can significantly shorten the incubation period of feeding rats (P<0.05), and increase the number of horizontal and vertical activities (P<0.01); the 5-HT and NE levels in serum were significantly elevated (P<0.05, P<0.01); the expressions of PI3K, AKT in hippocampal increased; the expression of SGK1 decreased (P<0.05, P<0.01). Conclusion Baishile Capsules can alleviate the depression like behavior in rat models, and regulate key factors of hippocampal PI3K signaling pathway, so as to exert antidepressant effects.

Objective To investigate the effects of Zuogui Jiangtang Jieyu Prescription on neurotrophic effects of astrocytes in diabetes mellitus rats with depression. Methods The diabetes mellitus with depression rat models were established by composite method, and then were randomly divided into 5 groups: model group, positive group, Zuogui Jiangtang Jieyu Prescription high-, medium-, low-dose groups, with 12 rats in each group. 12 normal rats were set as control group. Each administration group was given relevenat medicine for gavage for continuous 4 weeks. Open-field test was used to evaluate the depression-like behavior. The expression of GFAP in astrocyte and MAP2 in neuron were tested by immunohistochemistry. The expression of protein and mRNA of BDNF, GDNF, and NGF were tested by Western blot and RT-PCR. Results Compared with the control group, the activities of rats in model group were significantly reduced. The expression of GFAP increased, while the expressions of MAP2, BDNF, GDNF and NGF decreased. Compared with the model group, the depression-like behavior of rats in model group were significantly improved. The expression of GFAP decreased, while the expressions of MAP2, BDNF, GDNF and NGF increased, and GFAP decrseaed significantly. Conclusion The secretion function of astrocyte can be improved by Zuogui Jiangtang Jieyu Prescription. Its anti-depression and neuron-protection function might be correlated with the enhancement of neurotrophic effects of astrocytes.

The technique of liquisolid compress is a new technique developed in 1990s, which was considered to be the most promising technique to improve the dissolution of water-insoluble drugs. In this article, tanshinone II(A) and the extracts of the ester-solubility fractions were chosen as the model drugs to evaluate the effects of the liquisolid technique for enhancement of dissolution properties of tanshinone II(A). Several liquisolid tablets (LS) formulations containing different dosage of drugs and various liquid vehicle were pre-pared and for all the formulations, microcrystalline cellulose and silica were chosen as the carrier and coating materials to evaluate their flow properties, such as angle of repose, Carr's compressibility index and Hausner's ratio. The interaction between drug and excipients in prepared LS compacts were studied by differential scanning calorimetry(DSC) and X-ray powder diffraction (XRPD). The dissolution curves of tanshinone II(A) from liquisolid compacts were investigated to determine the technique's effect in improving the dissolution of tanshinone II(A) and its impacting factors. According to the results, the dissolution increased with the rise in the dissolution of the liquid-phase solvent. The R-value and drug dosage can significantly affect the drug release, but with less impact on active fractions. This indicated that liquisolid technique is a promising alternative for improvement of dissolution property of water-soluble drugs, and can make a synergistic effect with other ester-soluble constituents and bettern improve the release of tanshinone II(A). Therefore, the technique of liquisolid compress will have a better development prospect in traditional Chinese medicines.
Calorimetry, Differential Scanning ; Diterpenes, Abietane ; chemistry ; Solubility ; X-Ray Diffraction

During the dilute acid pretreatment of lignocellulosic materials such as corn stover, hemicellulose is hydrolyzed into monosaccharides, and meanwhile, toxic by-products are simultaneously generated, which may influence ethanol fermentation thereafter. Studies on the inhibitory effects of the by-products on ethanol fermentation are of practical use for further improvement of ethanol yield from lignocellulosic materials. Five by-products, including acetic acid, formic acid, vanillin, furfural and 5-hydroxymethylfurfural, were identified to be the main components in the hydrolysate of dilute acid pretreatment of local corn stover, which were added into the medium at different concentrations to study their impacts on the growth and ethanol fermentation of a recombinant xylose-utilizing yeast strain, S. cerevisiae 6508-127. The ethanol production was inhibited by formic acid and acetic acid to a lesser extent than that to the growth, and formic acid was shown to be much more toxic than acetic acid, showing severe inhibitory effects at the concentration of 1g/L, half of the concentration for acetic acid which showed remarkably negative effects on ethanol fermentation. Vanillin caused a much longer lag-phase in growth when the concentration was 2g/L, and the lag-phase was not obvious at lower concentrations. At the concentration of 6g/L, vanillin completely inhibited the fermentation as well as the cell growth. 5-Hydroxymethylfurfural was showed to remarkably inhibit ethanol production, but the biomass yield was higher by exogenous addition of 5-Hydroxymethylfurfural than control. Furfural at 0.5~1.5g/L inhibited the cell growth, but the ethanol yield was higher than that of the control experiment. It was also found that vanillin, furfural and 5-hydroxymethylfurfural could be assimilated and metabolized by S. cerevisiae 6508-127 under the experimental conditions.

In view of the predominance problems in present experimental teaching of medical microbiology, we reformed the experimental contents and experimental teaching methods,to establish an entire experimental curriculum teaching system to be suitable for seven-year system and long educational system students,which manifests the creative teaching idea.

Depression is a common emotional disorder that seriously affects people's life and health all over the world. The pathogenesis of depression is complex, and traditional Chinese medicine (TCM) for antidepressants has a good therapeutic effect because of its multi-component, multi-pathway, and multi-target action mode. At present, the anti-depressive mechanism of TCM has not been fully clarified, but it is clear that depression is closely related to metabolic health. Therefore, in order to further explore the anti-depressive mechanism of TCM, this paper proposes research strategies on the anti-depressive mechanism of TCM based on functional metabolomics from the perspective of metabolism, the potential biomarkers of depression are analyzed with the help of multi-omics combined analysis technology, and the functional molecules of TCM for antidepressant are studied. Molecular biology techniques are used to accurately capture the molecular interactions between biomarkers of depression and functional compounds, which identify effective drug targets and further elucidate the biochemical functions and related mechanisms involved in depression metabolic disorders. This paper systematically reviews the research strategies and applications of functional metabolomics in the anti-depressive mechanisms of TCM, expounds on the core value of functional metabolomics, and summarizes the current research status and hot issues of TCM for antidepressants in recent years, providing new methods and new ideas for the study of mechanisms of TCM with the help of functional metabolomics.

Objective To explore the residual undifferentiated mouse embryonic stem cells (ESCs) in embryoid bodies. Methods Mouse R1 and Oct-4-GFP transgenic ESCs were firstly cultured in suspension to form embryoid bodies (EBs). Twenty days later, EBs were digested into single cells and then re-plated in standard ESC culture condition. The morphology of residual undifferentiated cells in EBs was observed, and surface makers and in vitro redifferentiation potency of residual cells were examined by flow cytometry and immunofluoreseent staining. The residual cells were expanded and subcutaneously injected into nude mice, and the specimens were harvested from the injection site for histological analysis 6 weeks after injection. Results There were residual undifferentiated ESCs in EBs differentiated for 20 days, which displayed clonal morphology and expressed undifferentiated cell markers of ESCs, including SSEA1, CD31, CD9 and Oct-4. The cells could be differentiated to form EBs again, and could be re-expanded from secondary EBs. The residual cells were able to form teratoma at the injection site, and mature endoderm, mesoderm and ectoderm tissues could be found in teratoma tissues. Conclusion There are residual undifferentiated ESCs after differentiation of ESCs into EBs. The residual ESCs can differentiate again in vitro and in vivo, and can residue again in the in vitro differentiation.