Mechanism of colon cancer cell apoptosis induced by telocinobufagin: role of oxidative stress and apoptosis pathway / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of telocinobufagin on viability and apoptosis of colorectal cancer (CRC) cells and explore the mechanism of telocinobufagin-induced apoptosis.</p><p><b>METHODS</b>MTT assay was performed to detect the viability of CRC cells exposed to telocinobufagin. Nuclear staining with Hoechst 33342 and flow cytometry were used to analyze the cell death of CRC cells. Expressions of proteins related with cell apoptosis and oxidative stress were determined with Western blotting.</p><p><b>RESULTS</b>Telocinobufagin decreased the viability of CRC cells in a time- and dose-dependent manner. The presence of karyopycnosis and apoptotic bodies together with the results of flow cytometry suggested that telocinobufagin induced cell apoptosis to cause cell death. Western blotting showed that telocinobufagin exposure of the cells resulted in upregulated p53 and Bax protein expressions and promoted cleavage of caspase 9 and PARP. Telocinobufagin induced phosphorylation of Bad and PARP cleavage, and suppressed phosphorylation of IKBα and TAK1 and expression of survivin in the cells.</p><p><b>CONCLUSION</b>Telocinobufagin can decrease the viability of CRC cells by inducing cell apoptosis, which involves p53-mediated Bax activation and inhibition of the IAP pathway.</p>

Pi (Spleen)-deficiency syndrome in tumor microenvironment is the pivotal pathogenesis of colorectal cancer immune escape / 中国结合医学杂志

Cancer immunoediting consists of three sequential phases: elimination, equilibrium, and escape. For colorectal adenoma-carcinoma sequence, the adenoma dysplastic progression may represent an equilibrium phase and the cancer stage as escape phase. Immune system eliminates transformed enterocytes by destroying them at first, sculpts them at the same time and selects the variants subsequently that are no longer recognized and insensitive to immune effectors, and finally induces immunosuppressive state within the tumor microenvironment that facilitates immune escape and tumor outgrowth. Immunosuppression and inflammation are the two crucial features of Pi (Spleen)-deficiency. Classic quotations, immune evidence and clinical observations suggest that Spleen (but not other organs) deficiency is the key pathogenesis of colorectal cancer (CRC) microenvironment. Weakness of old age, immunosuppressive cytokines from chronic inflammation, tumor-derived immunosuppressive factors and surrendered immune cells-regulatory T cells, myeloid-derived suppressor cells and tumor associated macrophages (TAMs) constitutes CRC microenvironment of Pi-deficiency. Furthermore, excess in superficiality, such as phlegm stagnation, blood stasis and toxin accumulation are induced by chronic inflammation on the basis of asthenia in origin, an immunosuppressive state. Great masters of Chinese medicine emphasize that strengthen Pi is the chief therapeutic principle for CRC which receives good therapeutic effects. So, Pi-deficiency based syndrome is the pivotal pathogenesis of tumor microenvironment. The immunosuppressive microenvironment facilitates immune escape which play an important role in the transition from adenoma to adenocarcinoma. There are some signs that strengthen Pi based treatment has potential capacity to ameliorate tumor environment. It might be a novel starting point to explore the mechanism of strengthen Pi based therapy in the prevention and treatment of CRC through regulation of tumor environment and immunoediting.

Surgical treatment for chronic achilles tendon rupture and severe scarring / 中国骨伤

<p><b>OBJECTIVE</b>To evaluate the clinical efficacy of autologous semitendinosus and gracilis tendon grafting with anchor repair for the treatment of chronic achilles tendon rupture and severe scarring.</p><p><b>METHODS</b>From April 2010 to October 2012,26 patients with chronic achilles tendon rupture(with Myerson type III ) and severe scarring were treated with autologous semitendinosus and gracilis tendon grafting with anchor repair. There were 19 males and 7 females,with an average age of 32 years old (ranged, 22 to 47 years). The time from injury to surgery was from 3 to 12 months (7 months on average). The plantar flexion strength of all injuried feet attenuated and single heel rise test were positive in 26 cases before operation. Plaster immobilization and routine rehabilitation therapy were performed after operation. Clinical effects were evaluated by Arner-lindholm criterion and complications were observed after operation.</p><p><b>RESULTS</b>All the patients were followed up from 12 to 24 months with a mean of 16 months. No complications such as achilles tendon re-rupture, wound infection, etc were found during follow-up period. According to the Arner-Lindholm standard, 15 cases got excellent results and 11 good.</p><p><b>CONCLUSION</b>Using autologous semitendinosus and gracilis tendon grafts with anchor repair to treat chronic achilles tendon rupture and severe scarring is a perfect surgical procedure.</p>

Effect of curcumin on radiosensitization of CNE-2 cells and its mechanism / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of curcumin (Cur) on radiosensitivity of nasopharyngeal carcinoma cell CNE-2 and its mechanism.</p><p><b>METHOD</b>The effect of curcumin on radiosensitivity was determined by the clone formation assay. The cell survival curve was fitted by Graph prism 6. 0. The changes in cell cycle were analyzed by flow cytometry (FCM). The differential expression of long non-coding RNA was detected by gene chip technology. Part of differentially expressed genes was verified by Real-time PCR.</p><p><b>RESULT</b>After 10 micro mol L-1 Cur had worked for 24 h, its sensitization enhancement ratio was 1. 03, indicating that low concentration of curcumin could increase the radiosensitivity of nasopharyngeal carcinoma cells; FCM displayed a significant increase of G2 phase cells and significant decrease of S phase cells in the Cur combined radiation group. In the Cur group, the GUCY2GP, H2BFXP, LINC00623 IncRNA were significantly up-regulated and ZRANB2-AS2 LOC100506835, FLJ36000 IncRNA were significantly down-regulated.</p><p><b>CONCLUSION</b>Cur has radiosensitizing effect on human nasopharyngeal carcinoma CNE-2 cells. Its mechanism may be related to the changes in the cell cycle distribution and the expression of long non-coding IncRNA.</p>

Chemical constituents of leaves of Psidium guajava / 中国中药杂志

To study the chemical constituents of the 95% ethanol extract of Psidium guajava. Compounds were separated by using a combination of various chromatographic methods including silica gel, D101 macroporous resin, ODS, Sephadex LH-20 and preparative HPLC. Their structures were elucidated by physicochemical properties and spectral data Eighteen compounds were isolated and identified as (+) -globulol (1), clovane-2beta, 9alpha-diol (2), 2beta-acetoxyclovan-9alpha-ol (3), (+) -caryolane-1 ,9beta-diol (4), ent-T-muurolol (5), clov-2-ene-9alpha-ol (6), isophytol (7), tamarixetin (8), gossypetin (9), quercetin (10), kaempferol (11), guajaverin (12), avicularin (13), chrysin 6-C-glucoside (14), 3'-O-methyl-3, 4-methylenedioxyellagic acid 4'-O-beta-D-glucopyranoside (15), p-hydroxy-benzoic acid (16), guavinoside A (17) and guavinoside B (18). Compounds 2-9 and 14-16 were isolated from this plant for the first time. The ethanol extract showed 61.3% inhibition against the proliferation of colon cancer cell line SW480.

Chinese integrative medicine: translation toward person-centered and balanced medicine / 中国结合医学杂志

Chinese integrative medicine (CIM) focuses on the integration of conventional medicine (biomedicine) with Chinese medicine (CM). Although the CIM field has witnessed several advancements, the definition and classification of CIM is not quite clear, given that an independent theory system has not yet been established in this field. Therefore, future research and studies should focus on the following objectives: (1) emphasizing CM features, (2) improving CIM positioning, and (3) establishing CIM standards. These concerted efforts will help CIM be at par with international standards and criteria. With the development of CIM, the world will embrace a new medical system providing person-centered treatment with a balanced medicine approach.

Dingxin recipe ( ) prevents ischemia/reperfusion-induced arrhythmias via up-regulating prohibitin and suppressing inflammatory responses / 中国结合医学杂志

<p><b>OBJECTIVE</b>To identify the underlying mechanisms of the protective effects of Dingxin Recipe (: , DXR), a Chinese compound prescription that has been used clinically in China for more than 20 years, on ischemia/reperfusion (I/R)-induced arrhythmias in rat model.</p><p><b>METHODS</b>A total of 30 rats were randomly divided into three groups: sham group, I/R group, and DXR-pretreated I/R (DXR-I/R) group. Rats in the DXR-DXRI/R group were intragastrically administrated with DXR (12.5 g/kg per day) for consecutive 7 days, while rats I/in the sham and I/R groups were administrated with normal saline. Arrhythmias were introduced by I/R and electrocardiograms (ECG) were recorded. Two-dimensional (2-D) polyacrylamide gel electrophoresis and matrix-matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) were used to identify assisted differentially expressed proteins. Immunohistochemistry, real-time quantitative polymerase chain reaction (RQ-RQPCR), Western blot, and enzyme-linked immunosorbent assay (ELISA) were performed to analyze proteins PCR), obtained in the above experiments.</p><p><b>RESULTS</b>DXR significantly reduced the incidence and mean duration of ventricular tachycardia and ventricular fibrillation and dramatically decreased the mortality, as well as arrhythmia score, compared with those of the I/R group. Among successfully identified proteins, prohibitin (PHB) and heart fatty acid binding protein (hFABP) were up-regulated in DXR-pretreated I/R rats compared with those of the I/R rats. In addition, compared with the I/R group, the level of glutathione (GSH) was elevated accompanied by reduced expressions of interleukin-6 (IL-6) and neutrophil infiltration in I/R rats with DXR pretreatment.</p><p><b>CONCLUSIONS</b>DXR could alleviate I/R-induced arrhythmias, which might be related to increased expression of PHB. The enhanced expression of PHB prevented against the depletion of GSH and consequently inhibited apoptosis of cardiomyocytes. Furthermore, up-regulation of PHB might ameliorate I/R-induced cell death and leakage of hFABP by suppressing neutrophil infiltration and IL-6 expressions.</p>

A study of 22 displaced intraarticular calcaneal fractures using locking plates with and without bone graft / 中国骨伤

<p><b>OBJECTIVE</b>To analyze locking plates with bone graft fusion in treating displaced intraarticular calcaneal fractures and determine whether it is beneficial in maintaining restoration of calcaneal height and anatomic reduction of the articular surface.</p><p><b>METHODS</b>From January 2007 to January 2008, 22 patients with displaced intraarticular calcaneal fractures were treated with locking plates with and without bone graft (divided into the bone graft group and non-bone graft group). There were 17 males and 5 females, ranging in age from 18 to 59 years with the mean of 35 years. Sanders III was in 14 cases and Sanders IV in 8 cases. Autologous iliac bone filled defects with locking plates fixation for the bone graft group; just locking plates fixation were performed for non-bone graft group. The Böhler angle and Gissane angle were measured before and after operation. The foot function of two groups were compared according to Maryland standard at the 6th month, 1, 2 years after operation.</p><p><b>RESULTS</b>All patients were followed up with an average of 25 months. There was no significant difference in the recovery of Böhler angle and Gissane angle between two groups (P > 0.05). After the 6 months,1, 2 years, there was no significant difference in the foot function between two groups (P > 0.05), in bone graft group, excellent result was in 6, 7 ,7 cases respectively; and in non-bone graft group, excellent results in 5, 6, 7 cases respectively.</p><p><b>CONCLUSION</b>Bone graft in the surgical treatment of calcaneal fractures is not an advantage.</p>

Effect of dachengqi decoction on expressions of TLR4 and TNF-alpha in the lung and the large intestine of mice with endotoxemia / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of Dachengqi Decoction (DCQD) on the key signaling molecules in lung and large intestine of mice with endotoxemia for exploring the exterior-interior relation between Fei and Dachang.</p><p><b>METHODS</b>A total of 32 BALB/c mice were randomly and equally allocated into four groups: mice in Group C and D were made into endotoxemia model by intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS); while those in Group A and B were not modeled but given intraperitoneal injection of saline instead. Thirty min after then, saline to Group A and C, and DCQD to Group B and D were given by gastric infusion, and mice were sacrificed 6 h later. Their tissues of lung and large intestine were taken for observing pathological changes by inverted microscopy with HE staining; detecting expressions of tumor necrosis factor-alpha (TNF-alpha) by LiquiChip system; determining gene transcription and protein expression of toll-like receptor 4 (TLR4) using reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, and the correlation of TNF-alpha and TLR4 levels in the lung and the large intestine tissue was analyzed. RESULTED: Compared with Group C, hemorrhage, pathologic toxemic features, including pulmonary edema and inflammatory cell infiltration as well as intestinal wall congestion and neutrophil infiltration, were significantly alleviated in Group D. Levels of TNF-alpha expression, TLR4 protein expression and gene transcription raised significantly in the modeled mice (P < 0.01), but comparison between the two modeled groups showed that the three parameters were lower in Group D than in Group C (P < 0. 01 or P < 0.05) respectively. Correlation analysis showed that the levels of TNF-alpha expression, TLR4 protein expression and gene transcription in pulmonary tissues were positively correlated with those in large intestinal tissues respectively (r = 0.973, P < 0.01, r = 0.906, P < 0.01, and r = 0.880, P < 0.01).</p><p><b>CONCLUSIONS</b>The effects of DCQD in alleviating pulmonary and large intestinal inflammation induced by endotoxemia might be correlated to its reduction on levels of TNF-alpha expression, TLR4 protein expression and gene transcription. Levels of the three parameters in the lung are correlated with the corresponding levels in the large intestine, which suggested the existence of exterior-interior relation between Fei and Dachang.</p>

Effects of acute cerebral ischemia on cerebral perfusion: quantitative evaluation by contrast-enhanced ultrasound in dogs / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the changes of cerebral blood flow (CBF) with real-time contrast-enhanced ultrasound (CEU) in a canine model of acute cerebral ischemia.</p><p><b>METHODS</b>Cerebral perfusion was assessed in 6 dogs subjected to craniotomy with CEU at the time of 0, 30, 60, 90 and 120 min after occlusion of the left common carotid artery (LCCA). The microvascular volume (A) and blood flow velocity (beta) in the brain were measured from the time-versus-acoustic intensity plots, and the value of Axbeta were calculated. 99mTc-ECD brain single photon emission computed tomography (SPECT) was performed on the day before the experiment and at 120 min after LCCA occlusion. The radioactive counts on both sides of the cerebral cortex were calculated.</p><p><b>RESULTS</b>A significant correlation was found between Axbeta from CEU and volume of the blood flow of the CCA from Doppler flowmetry. A, beta and Axbeta values varied significantly between the different time points (P>0.001). The ipsilateral hemisphere showed a low-perfusion state while the contralateral hemisphere showed a high-perfusion state immediately after the occlusion.</p><p><b>CONCLUSIONS</b>The changes of beta is the main regulation mechanism during acute cerebral ischemia in dogs.</p>

Effects of sophoridine on the growth and expressions of p53 and vascular endothelial growth factor of transplanted solid tumor SW480 in nude mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effects of sophoridine on the transplanted solid tumor SW480 and the expression of p53 and vascular endothelial growth factor (VEGF) in the tumor in nude mice.</p><p><b>METHODS</b>The nude mouse model bearing transplanted solid tumor SW480 was established, and the changes in the volume and weight of the tumor were determined after treatment of the mice with sophoridine. Western blotting and immunohistochemistry were employed to examine the expressions of p53 and VEGF proteins, respectively, and fluorescence quantitative PCR used to detect their mRNA expressions in the tumor tissue.</p><p><b>RESULTS</b>The volume and weight of the tumor xenograft in sophoridine group decreased in comparison with those in the control group. Sophoridine treatment resulted in lowered expressions of p53 and VEGF at both the protein and mRNA levels in the tumor explants as compared with the control group, with a tumor inhibition rate of 34.07% in nude mice.</p><p><b>CONCLUSION</b>Sophoridine can inhibit the growth of transplanted solid tumor of human SW480 cell line, the mechanism of which involves the inhibition of p53 and VEGF expression.</p>

Proteomic analysis of chronic restraint stress-induced Gan (肝)-stagnancy syndrome in rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To analyze the proteomic characteristics of Gan (肝)-stagnancy syndrome (GSS) by seeking the differential protein in blood and tissues of GSS model rats.</p><p><b>METHODS</b>GSS model rats were established by chronic restraint stress, keeping rats in restrain chamber for 6 h every day for 21 successive days. Their blood and liver samples were collected at the end of experiment for differential protein detection with methods of isoelectrofocusing and polyacrylamide SDS-PAGE, silver staining, and scanning. The gel images were analyzed with Imagemaster 2D Elite software, and the excavated differential protein spots were identified with matrix assistant laser resolving TOF mass spectrometry, Western blot, ELISA, and RT-PCR, respectively.</p><p><b>RESULTS</b>A method for isolating the protein in blood serum and tissues by two-dimensional gel electrophoresis was established and optimized. Six serum proteins and three liver proteins that differentially expressed were identified. The down-regulated differential proteins in serum of GSS model rats were serum albumin precursor, beta 1 globin, antibody against muscle acetylcholine receptor, Ig lambda-2 C region, and transthyretin (TTR), and those in liver tissue were aryl sulfotransferase, enoyl-CoA hydratase, and TTR. TTR down-regulation was found in both serum and liver. Preliminary biological information analysis showed that these differential proteins involved in immune, neuroendocrine, nutrition, and substance metabolism.</p><p><b>CONCLUSION</b>Proteomic analysis of differential proteins showed that TTR, aryl sulfotransferase, and enoyl-CoA hydratase expressions are downregulated in the GSS model rats, suggesting that the susceptibility of cancer could be enhanced by chronic stress.</p>

Changes in the hemodynamics of rats with immunological liver fibrosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the changes in the hemodynamics of rats with immunological liver fibrosis and explore the pathogenesis of "blood stasis" in liver fibrosis.</p><p><b>METHODS</b>Rat models of liver fibrosis were established by multiple intraperitoneal injections of pig serum. The hematocrit, blood viscosity at the shear rate of 150/s, 30/s, 5/s, and 1/s, serum markers for liver fibrosis, and serum transaminase levels were measured in the control and model rats.</p><p><b>RESULTS</b>The hematocrit, blood viscosity at different shear rates, hyaluronic acid (HA), laminin (LN), procollagen type III (PCIII), type IV collagen (CIV), glutamic-pyruvic transaminase (ALT) and glutamic-oxaloacetic transaminase (AST) increased significantly in the rats with experimental liver fibrosis appeared as compared with those in the control rats. Positive correlations were noted between blood viscosity at different shear rates and serum concentrations of the fibrosis markers (HA, LN, PCIII, and CIV) in the model rats.</p><p><b>CONCLUSION</b>The changes in the hemodynamics in rats with immunological liver fibrosis suggests the role of "blood stasis" in the pathogenesis of liver fibrosis and provide experimental evidence for therapies to "activate the blood circulation and dissipate blood stasis" for treatment of liver fibrosis.</p>

High-performance capillary electrophoresis for determining the chemical compositions of Gegen and Fenge extracts / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop a method for analyzing the chemical compositions of Gegen and Fenge extracts using high-performance capillary electrophoresis (HPCE).</p><p><b>METHODS</b>Using HPCE/DAD, the chemical composition of the extracts was analyzed with the buffer solution of 40 mmol/L borax containing 16.7% methanol, with injection pressure at 137.9 kPa for 5 s, separation voltage at 25 kV in 0-5 min time range and at 22 kV in 5-25 min time range, and the temperature of the capillary of 20 degrees celsius.</p><p><b>RESULTS AND CONCLUSION</b>The method for analysis of Gegen and Fenge extracts was established, which identified puerarin and daidzein as the two major components. This simple and rapid analysis method can be used for Gegen and Fenge extract fingerprinting.</p>

Effects of bagasse polysaccharide on the immune functions of immunosuppressed mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effects of bagasse polysaccharide on the immune functions of immunosuppressed mice.</p><p><b>METHODS</b>Immunosuppressed mouse models were established by intraperitoneal injections with cyclophosphamide followed by daily intragastric administration of bagasse polysaccharide. After the treatments, the mice were examined for immune organ weight index, phagocytotic function of the macrophages, delayed type hypersensitivity, serum IgM level following exposure to chicken red blood cells, formation of hemolytic plaques, T cell percentage and lymphocyte transformation.</p><p><b>RESULTS</b>Treatment of the immunosuppressed mice with bagasse polysaccharide at the daily dose of 200 and 400 mg/kg significantly increased the weight of the immune organs, phagocytotic function of the macrophages, delayed type hypersensitivity, serum IgM level against chicken red blood cells, formation of hemolytic plaques, T cell percentage and lymphocyte transformation.</p><p><b>CONCLUSION</b>Bagasse polysaccharide can enhance the immune functions of immunosuppressed mice.</p>

Construction and identification of RNA interference vector for human tissue factor gene / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a RNA interference vector for human tissue factor (TF) gene.</p><p><b>METHODS</b>Human TF short hairpin RNA (shRNA) sequence was designed using online design software (Invitrogen) and synthesized into double-strand oligonucleotide (ds oligo), which was cloned into the pENTRTM/U6 plasmid, followed by transformation of the product into competent Top10 E. coli cells. After expansion of the transformed bacteria, the plasmid was extracted and sequenced, which was subsequently transfected into human umbilical vein endothelial cells (HUVECs). The interference effect of the vector on the target gene expression was detected by RT-PCR and immunofluorescence assay.</p><p><b>RESULTS</b>The sequencing result indicated that the plasmid pENTRTM/U6-RelB-shRNA was constructed correctly, which resulted in effective inhibition of TF expression in HUVECs after transfection.</p><p><b>CONCLUSION</b>The RNA interference vector against human TF gene has been constructed successfully, which may provide a stable transfection vector for potential treatment of blood coagulation abnormalities.</p>

Reconstruction of an intracellular transduction system based on HIV-1 TAT protein transduction domain / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the reasons for the low intracellular transduction efficiency of a previously constructed His-TAT-Flag recombinant protein and establish a more efficient transduction system.</p><p><b>METHODS</b>The Flag tag of pET14b-His- Tat-Flag vector was deleted with PCR mutant kit, and enhanced green fluorescent protein (EGFP) coding sequence was inserted into the new pET14b-His-TAT recombinant vector. Enzyme digestion and DNA sequencing were performed for identification of pET14b-His-TAT-EGFP vector, which was then transformed into E. coli BL21(DE(3)). After IPTG induction, the recombinant protein of His-TAT-EGFP was isolated and analyzed with SDS-PAGE. Purified His-TAT-EGFP recombinant protein was added to ECV304 cells and the fluorescence was observed to evaluate the transduction efficiency.</p><p><b>RESULTS</b>pET14b-His-TAT vector and pET14b-His-TAT- EGFP vector were successfully constructed, which was identified with enzyme digestion and DNA sequencing. His-TAT-EGFP fusion protein was expressed and purified successfully and showed cellular transduction activity.</p><p><b>CONCLUSION</b>The prokaryotic expression vector has been successfully constructed by modifying pET14b-His-TAT-Flag, and the expressed and purified recombinant protein of His-TAT-EGFP possesses high efficiency of intracellular transduction activity.</p>