MRF4 is one of muscle regulatory factors and plays critical roles during skeletal muscle development. The muscle development is important for the fish growth which is an important economic factor for the fish culture. To analyze the function of MRF4 in fish, the founder MRF4 antibody was prepared. The flounder MRF4 was cloned, ligated into prokaryotic expression vector pET-30b and expressed in strain E. coli BL21 (DE3). The recombinant flounder MRF4 fusion protein was soluble and purified with cobalt IMAC resins. To prepare MRF4 polyclonal antibodies, rabbits were immunized with the soluble protein and the increasing level of antibodies was determined by Western blot. Also, the endogenous flounder MRF4 was recognized by the anti-serum. The result further proved the existence of the anti-MRF4 antibody in the anti-serum, which will be useful for studies on the function of flounder MRF4.

Objective To explore the differences in behavior of anxiety individuals and the normal control under different feedback (positive feedback,negative feedback).Methods 400 university students were investigated using self-rating anxiety scale(SAS) and state anxiety inventory (SAI) and the data were analyzed with balloon analogue risk task.Results The score of anxiety group(24.28±6.12) was significantly lower (t=-2.25,P＜0.05) than that of control group(29.80±7.26) in the number of busting balloons.But in the number of busting balloons of positive and negative feedback condition,scores of anxiety group and control group were not significantly different(t=-1.82,P＞0.05;t=-1.78,P＞0.05).Conclusions Influence of anxiety on decision-making often manifests as risk aversion.

Objective To observe the effects of Jieyu-Chufan capsule(JCC) on the behavior and the monoamine neurotransmitter content of cerebral in depression mouse model and explore its mechanism. Methods The ICR male mouse were randomly divided into seven groups according to their weights(the model group, the low, middle and high doses group, Baiyoujie group, the amitriptyline group, the normal control group). The low, middle and high doses group were given JCC with 1.250, 2.500, 5.000 g/kg separately by intragastric administration. Baiyoujie group was given 0.014 g/kg Baiyoujie by gavage. The amitriptyline group was given 0.050 g/kg amitriptyline by gavage. The model group and the normal control group received the same volume of distilled water intragastrically. The drugs were administrated once a day. The brepharoptosis and movement conditions of mouse were observed after 22 days. The monoamine neurotransmitter content of each group was measured after 25 days. Results Compared to the model group, the number of mouse with brepharoptosis decreased in the high dose group after 1, 2, 6 hour with reserpine injection (P<0.05 or P<0.01);the number of mouse came out from the circle increased after 4 hours with reserpine injection (P<0.05 or P<0.01);the content of NA (880.45 ± 428.81 ng/g, 875.98 ± 449.33 ng/g vs. 299.92 ± 267.08 ng/g) and DA (2 305.99 ± 530.37 ng/g, 2 169.99 ± 278.19 ng/g vs. 1 439.34 ± 357.33 ng/g) in the middle and high dose group increased (P<0.01);the content of 5-HT (781.43 ± 135.10 ng/g vs. 492.01 ± 192.80 ng/g) in the middle group increased (P<0.01). Conclusions JCC showed an antidepression influence in depression mouse model induced by reserpine. The mechanism may be related to regulating the level of monoamine neurotransmitter in the nervous centralis .

ObjectiveTo investigate the effects of Saposhnikovia divaricata extract combined with arsenic trioxide (ATO) on the proliferation and apoptosis in chronic myelogenous leukemia K562 cells. MethodsSaposhnikovia divaricata extract was prepared.Cultured K562 cells were treated with different concentration of Saposhnikovia divaricataextract or/and ATO for 48h. Cell proliferation was determined using the MTT assay. Apoptosis and cell cycle were detected using flow cytometry.ResultsThe MTT assay showed that Saposhnikovia divaricata extract of 750,1 000,1 250,1 500 μg/ml had a significantly proliferation inhibitory effect compared with control group, the inhibitory rates were 23.29% ± 3.31%, 48.30% ± 2.50%, 79.62% ± 3.41% and 88.94% ± 0.06%, respectively (allP<0.05); Saposhnikovia divaricata extract of 500 μg/ml combined with ATO of 1.0, 0.5 μg/ml significantly increased inhibitor rates compared with ATO of 1.0, 0.5 μg/ml (64.99% ± 5.18%vs. 44.48% ± 3.31%,38.59% ± 3.88%vs.26.30% ± 5.03%; allP<0.05). FCM showed that Saposhnikovia divaricata extract of 500 μg/ml combined with ATO of 2.0, 1.0, 0.5 μg/ml significantly increased apoptotic rate compared with ATO group of 2.0, 1.0, 0.5 μg/ml (33.97% ± 0.59%vs.20.97% ± 2.17%, 13.53% ± 0.47%vs.9.77%±0.64%、6.63%±&0.40%vs.4.00%±0.46%; allP<0.05 ). Cell cycle results showed that Saposhnikovia divaricata extract of 500μg/ml combined with ATO of 2.0,1.0, 0.5μg/ml significantly increased the rate of S phase compared with ATO group of 2.0, 1.0, 0.5 μg/ml (60.25 ± 2.59%vs.55.61 ± 1.28%, 60.89 ± 1.53%vs.37.96 ± 1.02%, 47.76 ± 0.87%vs.39.90 ± 0.92%; allP<0.05).ConclusionsSaposhnikovia divaricataextract could obviously inhibit the proliferation of K562 cells and enhance the apoptotic effect of ATO. ATO could induce a G2/M phase arrest, while Saposhnikovia divaricata extract combined with ATO could induce a S phase arrest in K562 cells.

One new dicyclopeptide cyclo-(L-N-methyl Glu-L-N-methyl Glu) (1), together with one new natural dicyclopeptide cyclo-(L-methyl Glu ester-L-methyl Glu ester) (2), and two known dicyclopeptides cyclo-(L-methyl Glu ester-L-Glu) (3), and cyclo-(L-Glu-L-Glu) (4), were isolated from the aerial parts of Dianthus chinensis L. Their structures were determined by spectroscopic analyses and chemical methods.

Objective To study the effects of ginkgo biloba extract(GBE)on c-reactive protein(CRP) and tumor necrosis factor-α(TNF-α)in serum and alveolar lavage fluid(BALF)from rats with chronic obstructive pulmonary disease(COPD). Methods 90 rats were randomly divided into groups A,B,C,D,E and F. There were 15 rats in each group. The rat model of COPD were established in groups B,C,D,E and F. Groups C and D were given intraperitoneal injections with GBE from day l to day l4 and day 29 to day 42. Groups E and F weregiven intraperitoneal injections with erythromycin from day l to day l4 and day 29 to day 42. After the end of experi-ment ,the contents of CRP and TNF-α in serum and BALF were detected in all groups. Results The contents of CRP and TNF-α in the serum and the BALF were markedly lower in groups C,D,E and F than in group B (P<0.05);and the contents of CRP in the serum and the BALF and TNF-αin the BALF were lower in groups C, E and F than in group D(P<0.05). Conclusions GBE can inhibit the airway and systemic inflammatory response in COPD rats. Early intervention is more effective.

Objective To study the effect of black granule capsules(BGCs) on growth of transplanted mouse hepatocarcinoma cells, cell proliferation cycle and apoptosis.Methods KM mouse were subcutaneously inoculated with Hepatocarcinoma cells (H22) and randomly divided into the model control group, the positive control group, the low, medium and high does of BGC group after 24h. The positive control group received intraperitoneal injection with 30 mg/kg cyclophosphamide. Mice of BGC groups were intragastricaly with different dosage of BGC (400, 800, 1 600 mg/kg). The model control group received intragastricaly with normal saline. The drugs were administrated once a day for seven days. The tumor inhibition rate was calculated at 24 h after the last administration. Flow cytometry was used to detect changes of cell cycle and apoptosis in harvested H22 tumor cells.Results The group of high does showed significant inhibitory effect on the growth of transplanted H22 tumor cells withthe inhibitory rate 38.78% (male) and 43.57% (female). Compared with model control group, groups of different dosages decreased time of G0-G1 phase (58.06% ± 9.65%, 55.10% ± 5.89%, 61.36% ± 15.95%vs. 74.47% ± 2.63%), increased tiem of Sphase (33.96% ± 11.90%, 32.67% ± 4.04%, 33.89% ± 9.82%vs. 14.37% ± 4.92%), and increased the apoptosis rate (31.12% ± 1.85%, 31.89% ± 2.16%, 40.64% ± 0.55%vs.21.75% ± 2.64%).Conclusion BGC has antitumor effect on mouse hepatocarcinoma H22 tumor cells, and its mechanism was to regulate cell proliferation cycle and induce the apoptosis.

OBJECTIVETo study the chemical constituents from the fruits of Leonurus japonicus.

METHODThe compounds were extracted by 70% methanol, then isolated and purified by column chromatography on silica gel, RP-18, Sephedax LH-20 and HPLC. Their structures were determined on the basis of spectroscopic data and physicochemical property.

RESULTTwo 28-noroleanane-derived spirocyclic triterpenoids including a new acylated nortriterpenoid were isolated and identified as (17R)-19(18 --> 17)-abeo-2alpha,18beta,23-trihydroxy-3beta-E-feruloyoxy-28-norolean-12-ene (1) and phlomistetraol B (2).

CONCLUSIONCompound 1 is a new compound,named as leonujaponin A. Both compounds 1-2 are isolated for the first time from Leonurus plants.

Background/Aims: Esophagogastric junction outflow obstruction (EGJOO) is characterized by elevated integrated relaxation pressure (IRP) and preserved esophageal peristalsis. The clinical significance of EGJOO is uncertain. This study aim to describe the clinical characteristics of these patients and to find out potential parameters to predict patients’ symptom outcome. Methods: Consecutive patients who received high-resolution manometry examination in our hospital in 2013-2019 and met the diagnostic criteria of EGJOO were retrospectively included. Motility and reflux parameters as well as endoscopy and barium esophagogram results were studied and compared. Patients were also followed up to record their treatment methods and symptom outcomes. Results: A total of 138 EGJOO (accounting for 5.2% of total patients taking high-resolution manometry examination in our hospital) patients were included. Only 2.9% of these patients had persistent dysphagia. A total of 81.8% of EGJOO patients had symptom resolution during follow-up. Patients with persistent dysphagia had significantly higher upright IRP (16.6 [10.3, 19.8] vs 7.8 [3.2, 11.5]; P = 0.026) than those without. Upright IRP can effectively distinguished patients with persistent dysphagia (area under curve: 0.826; P = 0.026) using optimal cut-off value of 9.05 mmHg. Conclusion EGJOO patients with persistent dysphagia and higher upright IRP (median > 9.05 mmHg) needs further evaluation and aggressive management.

Background/Aims: Esophagogastric junction outflow obstruction (EGJOO) is characterized by elevated integrated relaxation pressure (IRP) and preserved esophageal peristalsis. The clinical significance of EGJOO is uncertain. This study aim to describe the clinical characteristics of these patients and to find out potential parameters to predict patients’ symptom outcome. Methods: Consecutive patients who received high-resolution manometry examination in our hospital in 2013-2019 and met the diagnostic criteria of EGJOO were retrospectively included. Motility and reflux parameters as well as endoscopy and barium esophagogram results were studied and compared. Patients were also followed up to record their treatment methods and symptom outcomes. Results: A total of 138 EGJOO (accounting for 5.2% of total patients taking high-resolution manometry examination in our hospital) patients were included. Only 2.9% of these patients had persistent dysphagia. A total of 81.8% of EGJOO patients had symptom resolution during follow-up. Patients with persistent dysphagia had significantly higher upright IRP (16.6 [10.3, 19.8] vs 7.8 [3.2, 11.5]; P = 0.026) than those without. Upright IRP can effectively distinguished patients with persistent dysphagia (area under curve: 0.826; P = 0.026) using optimal cut-off value of 9.05 mmHg. Conclusion EGJOO patients with persistent dysphagia and higher upright IRP (median > 9.05 mmHg) needs further evaluation and aggressive management.