ObjectiveTaking classical herbal pair compatibility research as the entry point， this study aimed to deeply investigate the material basis and compatibility rules underlying the antidepressant effects of the traditional Chinese medicine （TCM） formula Zhizi Houpotang， and to elucidate its antidepressant mechanism， with a particular focus on its regulation of neuroinflammatory responses mediated by the NOD-like receptor protein 3 （NLRP3）/gasdermin D （GSDMD） signaling pathway and the consequent improvement of neuronal synaptic plasticity. MethodsC57BL/6J mice were randomly divided into a blank control group， a chronic unpredictable mild stress （CUMS） depression model group， a Zhizi Houpotang full-formula group （6 g·kg^-1·d^-1）， a Magnoliae Officinalis Cortex （MOC）-Aurantii Fructus Immaturus （AFI） herbal pair group （4.2 g·kg^-1·d^-1）， a Gardeniae Fructus （GF）-MOC herbal pair group （4.2 g·kg^-1·d^-1）， a GF-AFI herbal pair group （3.6 g·kg^-1·d^-1）， and a positive drug group （fluoxetine， 12 mg·kg^-1·d^-1）. Depressive-like behaviors in mice were evaluated using behavioral tests. Immunofluorescence staining was used to label and quantify the expression of the microglial marker ionized calcium-binding adaptor molecule 1 （Ibal） and the purinergic receptor P2X ligand-gated ion channel 7 （P2RX7） in the prefrontal cortex （PFC）. Enzyme-linked immunosorbent assay （ELISA） was applied to detect the levels of inflammatory cytokines interleukin-1β （IL-1β） and interleukin-18 （IL-18） in serum and PFC tissues. Western blot was employed to determine the expression of pannexin 1 （Panx1）， P2RX7， NLRP3， apoptosis-associated speck-like protein containing a CARD （ASC）， Caspase-1， GSDMD， postsynaptic density protein 95 （PSD95）， and the presynaptic protein Synapsin 1 in PFC tissues. Golgi staining was used to assess dendritic spine density of neurons in the PFC. ResultsCompared with the blank control group， the depression model group exhibited significant depressive-like behaviors. In addition， the immunofluorescence areas of Ibal and P2RX7 in the PFC were significantly increased （P<0.01）， the levels of IL-1β and IL-18 in serum and the PFC were significantly elevated （P<0.01）， and the protein expression levels of Panx1， P2RX7， NLRP3， ASC， Caspase-1， and GSDMD in the PFC were significantly upregulated （P<0.01）. In contrast， the protein expression levels of PSD95 and Synapsin 1 were significantly downregulated （P<0.01）， and neuronal dendritic spine density was significantly reduced （P<0.01）. Compared with the model group， the Zhizi Houpotang full-formula group and the GF-MOC herbal pair group showed significant improvement in all the above indicators （P<0.01）. The GF-AFI herbal pair group improved all the above indicators except P2RX7， Caspase-1， GSDMD， and PSD95 （P<0.05， P<0.01）. In contrast， the MOC-AFI herbal pair group showed no statistically significant improvement in any of the above indicators compared with the model group. ConclusionZhizi Houpotang and its key herbal pair， GF-MOC， can effectively ameliorate CUMS-induced depressive-like behaviors in mice. Its core antidepressant mechanism may involve inhibition of P2RX7/Panx1 signaling， thereby blocking the NLRP3/GSDMD-mediated pyroptosis pathway and significantly reducing the release of inflammatory cytokines IL-1β and IL-18. Simultaneously， it upregulates the expression of synapse-related proteins PSD95 and Synapsin 1 and increases dendritic spine density， promoting the recovery of synaptic plasticity. These results suggest that GF plays a key role in the antidepressant effects of this formula， and that the compatibility of GF with MOC may represent the principal herbal pair combination responsible for its core therapeutic action.

【Objective】 To establish a dry fluorescent luminescence method for the detection of antibodies to hepatitis C virus (HCV) and evaluate its clinical application. 【Methods】 Anti-HCV antibody was detected by double-antigen sandwich dry fluorescent luminescence method established using multi-epitope chimeric antigen. The established method was used to detect national reference samples(positive 20, negative 20), and a total of 349 clinical samples, including 108 HCV patients, 36 patients with other diseases and 205 healthy individuals, which were tested in parallel with enzyme-linked immunoassay (ELISA) to evaluate the performance of the established method. 【Results】 The concordance rate of positive and negative(each 20) reference samples were both 100% (20/20), and the CV of precision reference sample was 9.16%, which met the requirements of national reference samples. In clinical performance evaluation, the AUC value was 0.984, and the sensitivity and specificity of the dry fluorescent luminescence method were 96.30% (104/108) and 96.27% (233/241). The overall concordance rate between dry fluorescent luminescence method and ELISA was 97.71% (341/349) (Kappa=0.952). 【Conclusion】 The dry fluorescence luminescence method of HCV antibody is simple and rapid, with high sensitivity and high specificity, and can be used in clinical application.

Betacoronavirus ; Coronavirus Infections ; Humans ; Pandemics ; Pneumonia, Viral ; Single-Cell Analysis

Aim To identify whether the petroleum e-ther fraction of Gardenia jasminoides Ellis ( GJ-PE ) could effetive exhibit a rapid antidepressant effect and also to investigate the biological mechanism. Methods Tail suspension test ( TST ) , forced swimming test ( FST ) and novelty suppressed-feeding ( NSF ) were used to screen the rapid antidepressant potential of ef-fective fractions of GJ-PE in KM mice at 24 h post a single administration. Tail suspension test ( TST) was also used at 30 min and forced swimming test ( FST ) was used at 2 h to test the initial onset time of effective fractions of GJ-PE in KM mice. Western blot was per-formed to examine the expression of BDNF and p-eEF2 in hippocampus of KM mice at 2 h and 24 h. Results An acute administration of GJ-PE1 decreased the im-mobility time of KM mice in FST at 2 h and 24 h and decreased the latency time in NSF at 24 h. GJ-PE3 de-creased the latency time in NSF at 24 h. GJ-PE4 in-creased the unit food consumption in NSF at 24 h. At 2 h post a single GJ-PE1 treatment, the expression of BDNF was significantly up-regulated while the expres-sion of p-eEF2 was significantly down-regulated. At 24 h post a single GJ-PE1 treatment, the expression of BDNF was significantly down-regulated while p-eEF2 expression was significantly up-regulated. Conclusion GJ-PE1 has the most significant rapid antidepressant potential among the four fractions of GJ-PE. The effec-tive time of GJ-PE1 is 2 h after drug treatment. The mechanism of the rapid antidepressant effect of GJ-PE1 at 2 h is related to the up-regulation of BDNF and down-regulation of p-eEF2 . GJ-PE3 and GJ-PE4 also have some features of rapid antidepressants. GJ-PE2 doesn′t have the rapid antidepressant potential.

Objective To investigate the correlations between eosinophil counts in induced sputum and lung function (FENO) and evaluate these parameters in medication adjustment in patients with asthma.Methods Sixty-five outpatients with mild to moderate persistent asthma ( mild,32 ; moderate,33 ) from January to August 2008 were enrolled in the study.All were treated with combined medications comprising inhaled corticosteroids plus long-acting β2 agonists for 1 year.Lung function (FEV1％ and PEF％ ),eosinophil counts in induced sputum,FENO,and Asthma Control Test (ACT) scores were obtained at regular follow-up intervals.Twenty-one healthy volunteers served as controls,and lung function,eosinophil counts in induced sputum,and FENO were also obtained.Results Sixty-three subjects completed 1-year or longer follow-up.Lung function of 63 subjects recovered quickly in the early days and improved slowly during the following 6 months.FENO decreased from (61 ± 25 ) nmol/L at baseline to ( 32 ± 19 ) nmol/L by the third month (q =7.32,P＜0.05) and to (22 ± 12) nmol/L by the sixth month,which showed significant difference from normal controls [ ( 13 ± 8) nmol/L; q =6.63,P ＜ 0.05 ].Eosinophil counts in induced sputum of the asthma group at baseline were (0.093 ±0.023) × 109/L and decreased to (0.032 ±0.011)× 109/L by the third month,which was significantly different from baseline and normal controls [ (0.005 ±0.003) × 106/ml; q =5.49,P ＜0.05 and q =5.87,P ＜0.05,respectively].FENO showed a significantly positive correlation with eosinophil counts in induced sputum in the first 6 months (r1 =0.612,r2 =0.558,r3 =0.675; all P＜0.05) and a negative correlation with FEV1 (r1 =-0.537,r3 =-0.658,r6 ＝ -0.623,r9 =-0.537,r12 =-0.597 ; all P ＜0.05 ) at any time point of the study.The ACT score of 63 subjects at baseline was 14 ±3,and the scores after treatment for 1,3,6,9,and 12 months were 18 ±5,19 ±7,23 ±2,24 ± 1,and 24 ± 1,respectively; at the same time,significant difference was found ( F =5.72,P ＜ 0.05).Effectiveness was found according to the ACT score only 1 month after treatment.Conclusion The parameters of FENO and eosinophil counts in induced sputum were sensitive in the detection of airway inflammation and may be useful in evaluation of the efficacy of treatment and adjustment of medication regimens.

Nonaqueous capillary electrophoresis is used for the determination of the contents of diosgenin and ruscogenin in Radix Ophiopogonis. The operating buffer was composed of 20 mmol x L(-1) Na2B4O7-HCl (pH 7.61) in 70% methanol. The applied voltage was 25 kV and detection potential was at +0.70 V. With these conditions, the components were successfully separated. The content of diosgenin in Radix Ophiopogonis was 0.018 mg x g(-1) and ruscogenin was 0.008 mg x g(-1). The average recoveries of diosgenin and ruscogenin were 102% and 99.2%, respectively. A new method of the quality control of diosgenin and ruscogenin in Radix Ophiopogonis is provided.

AIM: To investigate relationship between activity of matrix metalloproteinases - 2 ( MMP - 2, 72 kD) and invasion, metastasis of breast cancer. METHODS: Useing zymography and computer software assisted analysis, the activitive levels of MMP- 2 (72 kD) in tissues from breast cancer were measeured. RESULTS: Mean activitive levels of MMP- 272 kD (13.93 + 3.60) in breast cancer were lower than those in benign disease (21.43 + 8.31), P < 0.05. There was no difference (P > 0.05) in MMP - 2 62 kD + 72 kD of benign and malignant dis ease, but MMP - 262 kD ( 13.83 + 4.53) and MMP - 262 kD/62 kD + 72 kD (0.48) respectively were significantly higher in malignant disease (P < 0.01). It was also found that MMP- 262 kD/62 kD + 72 kD were apparently higher in invasive carcinomas (0.48) and lymph node metastases (0.61), P < 0.01, respectively. CONCLUSION: These results demonstrated that a clear relationship between MMP - 2 activity and the invasion and metastasis of breast carcinoma.

AIM: To investigate relationship between activity of matrix metalloproteinases-2 ( MMP-2, 72 kD) and invasion, metastasis of breast cancer. METHODS: Useing zymography and computer software assisted analysis, the activitive levels of MMP-2 (72 kD) in tissues from breast cancer were measeured. RESULTS: Mean activitive levels of MMP-2 72 kD (13.93?3.60) in breast cancer were lower than those in benign disease (21.43?8.31), P0.05) in MMP-2 62 kD+72 kD of benign and malignant disease, but MMP-2 62 kD (13.83?4.53) and MMP-2 62 kD/62 kD+72 kD(0.48) respectively were significantly higher in malignant disease (P

It was an important strategy in cancer gene therapy that transferring foreign gene to fibroblasts to express synthetic protein to exert antitumor effect. But most of studies regarded the foreign gene-modified fibroblasts as a by-stander, only a delivery source of synthetic protein, and neglected their irnmunological characters. In this paper, human primary dermal fibroblasts were modified by human IL-2 gene by retrovirus vector and induced by human IFN-?. The results showed that the MHC- I , MHC- II and CD40 expression on the surface of IFN-? induced fibroblasts were up-regulated significantly in comparison with those of non-induced cells. The IL-2, IL-1, IL-6 secretion were detected as a increased level in supematants of the IFN-?-induced, IL-2 gene modified fibroblasts. Because these molecules and cytokines play important roles in antigen presentation and effector activation, it can be inferred that the IFN-y induced, IL-2 gene modified fibroblasts could be used as antigen presenting cells in addition to delivery cells to activated the effector cells.

Adenoviruses harboring E. coli cytosine deaminase gene (AdCD) were used to transfect murine FBL-3 ery-throleukemia cells in vitro. FBL3 cells infected with AdCD were more sensitive to 5-fluorocytosine (5-FC) than cells infected with a control adenovirus AdLacZ. Further study indicated that this combination therapy (AdCD and 5-FC) killed tumor cells by inducing apoptosis of FBL-3 cells. The supematants from FBL-3 cells treated with AdCD/5-Fc were transferred on the culture system of uninfected (wild - type) FBL-3 cells, the result indicated that only 6.25% of the supernatant could induce significant cytotoxicity on wild type FBL3 cells. The results demonoustrated that bystander effect plays an important role in AdCD-mediated cytotoxicities. Direct injection of AdCD into established subcutaneous FBL3 tumor in mice followed by daily intraperitoneal injection of 5-FC for 10 days was found to inhibit tumor growth significant-