Hepatocellular carcinoma (HCC) is a serious threat for human health, the incidence of HCC in China accounts for more than 50% worldwide. There is an urgent need to develop novel anticancer agents for the treatment of HCC patients. Here we characterized the inhibitory effect and the molecular mechanism of protopine on HCC cancer cells. The results of a CCK-8 assay indicated that protopine displays anticancer activities on HCC cells. Flow cytometry and JC-1 staining confirmed that treatment with protopine decreased the mitochondrial membrane potential and induced apoptosis in HCC cells. Western blot analysis showed that protopine was able to increase protein expression in the mitochondrial apoptotic pathway; the level of cytochrome C, apoptotic protease activating factor-1 (Apaf-1), Bax, cleaved-poly ADP-ribose polymerase (cleaved-PARP), cleaved-caspase-3, and cleaved-caspase-9 were increased while the expression of Bcl-2 was suppressed significantly. An in vivo study revealed that protopine significantly suppressed the growth of tumors in nude mice bearing HepG2 cells. Administration of protopine intraperitoneally at a concentration of 50 mg·kg^-1 inhibited tumor growth by 72.46%. Animal experiments were carried out according to the Regulation of the Animal Ethics Committee of Southwest Medical University. This study provides preliminary evidence that there is potential to develop protopine as a lead compound for the treatment of HCC.

The β-Glucuronidase gene (sbGUS) cDNA firstly from Scutellari abaicalensis leaf was cloned by RT-PCR, with GenBank accession number KR364726. The full length cDNA of sbGUS was 1 584 bp with an open reading frame (ORF), encoding an unstable protein with 527 amino acids. The bioinformatic analysis showed that the sbGUS encoding protein had isoelectric point (pI) of 5.55 and a calculated molecular weight about 58.724 8 kDa, with a transmembrane regions and signal peptide, had conserved domains of glycoside hydrolase super family and unintegrated trans-glycosidase catalytic structure. In the secondary structure, the percentage of alpha helix, extended strand, β-extended and random coil were 25.62%, 28.84%, 13.28% and 32.26%, respectively. The homologous analysis indicated the nucleotide sequence 98.93% similarity and the amino acid sequence 98.29% similarity with S. baicalensis (BAA97804.1), in the nine positions were different. The expression level of sGUS was the highest in root based on a real-time PCR analysis, followed by flower and stem, and the lowest was in stem. The results provide a foundation for exploring the molecular function of sbGUS involved in baicalcin biosynthesis based on synthetic biology approach in S. baicalensis plants.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Computational Biology ; Glucuronidase ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary ; Scutellaria baicalensis ; chemistry ; enzymology ; genetics ; Sequence Alignment

ObjectiveTo observe the effects of acupuncture combined with massage on shoulder subluxation and motor function recovery of the upper extremities in patients with hemiplegia after stroke.Methods60 hemiplegic patients were randomly divided into two groups for treatment with acupuncture cooperates with massage or usual rehabilition, respectively. The recovery of the patient's shoulder subluxation and movement function of upper extremities were evaluated 2 months after treatment. ResultsThe shoulder subluxation and movement function of the upper extremities were improved after treatment with both therapies (P<0.01), and acupuncture cooperates with massage showed better effect (P<0.01). ConclusionAcupuncture cooperates with massage can facilitate the recovery of shoulder subluxation and motor function of the upper extremities after hemiplegia.

Objective:To observe the clinical efficacy of Ba-pulling and Qian-traction manipulation with neck suspension and movement for acute cervical radiculopathy. Methods: A total of 85 patients who met the inclusion criteria were randomized into an observation group and a control group by random numbers, with 43 cases in the observation group and 42 cases in the control group. The observation group was treated with Ba-pulling and Qian-traction manipulation with neck suspension and movement;while the control group was treated with Bashen-pulling and stretching manipulation in a supine position. The treatment was performed once a day, 10 times as a treatment course. The therapeutic efficacy was evaluated after 1 treatment course, and the changes in the scores of visual analog scale (VAS) and neck disability index (NDI) were observed. Results: The total effective rate was 97.7% in the observation group, and 83.3% in the control group, and the difference between the two groups was statistically significant (P＜0.05). After treatment, the VAS and NDI scores of both groups were significantly decreased (both P＜0.01), and the differences in the VAS and NDI scores between the two groups were statistically significant (both P＜0.01). Conclusion: Both Ba-pulling and Qian-traction manipulation with neck suspension and movement and Bashen-pulling and stretching manipulation in a supine position can relieve pain and improve cervical function in patients with acute cervical radiculopathy, and Ba-pulling and Qian-traction manipulation with neck suspension and movement can produce more significant efficacy than Bashen-pulling and stretching manipulation in a supine position.

We reported a case of cryptogenic organizing pneumonia ( COP) who was admitted to the hospital in July 2008 and reviewed the Chinese literature of COP from 2003 to 2008. The most common symptoms of COP are fever, cough and exertional dyspnea. The imaging characteristics of COP are similar to those of pneumonia, therefore is often misdiagnosed as pneumonia with a high misdiagnosis rate. Lung biopsy is the main method for pathological diagnosis; polypoid growth of granulation tissue was noted within respiratory bronchioles, small airways and alveolar spaces.

Objective Gambogic acid ( GA) can suppress the growth of multiple tumor cells , including gastric carcinoma , hepatoma , hematologic neoplasms and breast carcinoma , but there have been few reports about its effect on urologic neoplasms .This study was to investigate the possible mechanisms of GA inducing bladder cancer cell apoptosis and cell cycle arrest . Methods We cultured human bladder cancer BIU8-7 cell lines in vitor and treated the cells in the logarithmic growth phase with isotonic saline solu-tion (negative control)or GA at the concentrations of 1.0, 2.0, and 3.0μmol/L, respectively.We determined the expression of the Caspase-3 protein in the tumor tissue using the immunohistochemical S-P method and detected GA-induced apoptosis of the bladder cancer cells and cell cycle changes by flow cytometry . Results The expressions of the Caspase-3 protein were 4.28 ±1.86, 5.03 ± 0.78, and 6.47 ±1.31 in the 1.0, 2.0, and 3.0μmol/L GA groups, respectively, significantly higher than 2.13 ±1.27 in the nega-tive control (P<0.05).Flow cytometry showed a gradual decrease of the cells in the G 0/G1 phase and a gradual increase in the G2/M phase , but no obvious change in the S phase . Conclusion Gambogic acid can promote the apoptosis , arrest the cell cycle , and in-hibit the proliferation of bladder cancer cells by increasing the expression of the Caspase -3 protein.

Objective To evaluate the roles of Toll-like receptor 2 (TLR2),TLR4 and myeloid differentiation factor 88 (MyD88) in immune recognition and immune mediation in sporotrichosis by detecting their expressions in skin lesions of sporotrichosis.Methods Biopsy specimens were obtained from the skin lesions of 19 patients with sporotrichosis and normal skin of 12 healthy human controls.Immunohistochemistry was performed to observe the expressions of TLR4 and MyD88,and real-time fluorescence-based quantitative PCR to quantify the expressions of TLR2 and MyD88 mRNAs.Data were expressed as mean ± standard error.Statistical analysis was done with the SPSS17.0 software.Independent samples t-test was conducted to compare the parameters between the lesional and control specimens.A p-value of less than 0.05 was considered to be statistically significant.Results TLR4 and MyD88 were mainly observed in the whole epidermis except the stratum corneum as well as plasma cells and lymphocytes in the dermis and subcutaneous tissue in lesional skin.However,TLR4 was nearly absent and MyD88 was weakly expressed in the dermis and subcutaneous tissue in normal skin.The expression levels of TLR4 and MyD88 were significantly higher in the lesional skin than in the control skin (TLR4,63.767 ± 3.829 vs.5.167 ± 3.246,t =4.82,P ＜ 0.05; MyD88,57.236 ± 4.744 vs.10.588 ± 1.640,t =3.30,P ＜ 0.05).Similarly,the lesional skin showed significantly stronger expressions of TLR2 and MyD88 mRNAs compared with the normal skin (TLR2,1.974 ± 1.452 vs.1.430 ± 1.073,P ＜ 0.05; MyD88,2.028 ± 2.061 vs.0.688 ± 0.422,P ＜ 0.05).Conclusion Sporothrix may induce the development of sporotrichosis by interacting with host immune system via TLR signaling pathways.

Objective To summarize the preliminary experience and early -stage follow -up results of simple transesophageal echocardiography(TEE)to guide right jugular internal vein for closure of atrial septal defect (ASD), and to investigate its feasibility.Methods A total of 24 ASD patients(12 male,12 female)treated by transcatheter ASD closure under solely guidance of TEE were summarized in Children Heart Center,People′s Hospital of Zhengzhou University from February 2015 to February 2016.Mean age,body weight and average diameter of the 24 cases of ASD were 5 -18 (12.4 ±5.6)years old,16 -60 (34.1 ±9.6)kg,5 -20 (11.0 ±2.5)mm,respectively.All patients were treated by right jugular internal vein transcatheter closure under solely guidance of TEE.The efficiency of imme-diate post -operation under TEE was estimated,and follow -ups were done at 24 hours,1 month,3,6 and 12 months after the procedures by transthoracic echocardiography,X -ray examination and electrocardiogram.Results A total of 23 patients were treated successfully under solely guidance of TEE.The diameter of devices was (14.5 ±3.6)mm.The procedural time was(20.5 ±7.2)min.One patient who had 2 ASD was treated by traditional thoracic surgery,because the small ASD had 5 mm residual shunt after the procedures.There were 3 cases with trivial residual shunt after the pro-cedures immediately.All patients could move at once after operation.The average follow -up time was(5.6 ±1.2) months.No patients suffered from residual shunt,occluder malposition,peripheral vascular injury,pericardial effusion or cardiac perforation during the follow -up.Conclusions For some selected patients,simple TEE guiding right jugular in-ternal vein for closure of ASD is applicable.The method not only prevents injury from radiation,but also shortens the in -bed time and avoid the lower limbs braking,with good early -stage follow -up results are desirable.

Objective To explore genes expression of adiponectin receptors during differentiation of SW872 preadipocytes. Met-(hods) SW872 preadipocytes were cultured in vitro and induced to differentiate by 0.6 mmol/L oleic acid. During the progress of diffe-(rentiation), the morphological changes of SW872 cells were observed and the differentiation rate was assayed by oil-red O staining. Adiponectin receptors mRNA was measured by reverse transcription-polymerase chain reaction during differentiation of SW872 preadipocytes. Results 1.After stimulated by 0.6 mmol/L oleic acid for 72 hours, almost all SW872 cells were differentiated,and there were lots of fat droplets in the cells.2.There were adiponectin receptors genes expressions in SW872 preadipocytes.After 72 hours,and the levels of adiponectin receptor(AdipoR) 1 mRNA and AdipoR 2 mRNA were markedly increased up to 2.54 and 4.09 times,respectively. Conclusion There are AdipoR1 and AdipoR2 genes expressions in fat cells and the expressions are differentiation-dependent.

Objective To study the effect of oleic acid on the differentiation of SW872 preadipocytes.Methods SW872 prea-(dipocytes) were cultured and induced to differentiate by 0.6 mmol/L oleic acid in vitro.After 24 h,48 h and 72 h of differentiation,the morphological changes of SW872 preadipocytes were observed and the differentiation rate was assayed by oil-red O staining.In addition,triglyceride(TG) mass was detected by chemical colorimetry methods.During the differentiation of SW872 preadipocytes,transcription factors including peroxisome proliferator activated receptor-?_2(PPAR-?_2) and CAAT/enhancer binding protein-?(C/EBP-?)(mRNA) were also measured by reverse transcription-polymerase chain reaction(RT-PCR).Results 1.SW872 preadipocytes were fibroblastic and had no obvious fat droplet in cytoplasm.However,when stimulated for 72 hby 0.6 mmol/L oleic acid,SW872 preadipocytes became more bigger and rounder and differentiated into mature adipocytes with lots of fat droplets in the cells.2.Compared with that of predifferentiation,the concentration of TG mass increased by 14 folds after 72-hour differentiation(P