Research progress in micro/nanobubbles for ultrasound diagnosis or treatment / 药学学报

In the past few decades, microbubbles were widely used as ultrasound contrast agents in the field of tumor imaging. With the development of research, ultrasound targeted microbubble destruction technology combined with drug-loaded microbubbles can achieve precise drug release and play a therapeutic role. As a micron-scale carrier, microbubbles are difficult to penetrate the endothelial cell space of tumors, and nano-scale drug delivery system—nanobubbles came into being. The structure of the two is similar, but the difference in size highlights the unique advantages of nanobubbles in drug delivery. Based on the classification principle of shell materials, this review summarized micro/nanobubbles used for ultrasound diagnosis or treatment and discussed the possible development directions, providing references for the subsequent development.

Application of PDX model in the evaluation of nano-delivery systems / 药学学报

Malignant tumor is a major disease affecting human health. The nano-delivery system itself has a unique size effect and it can achieve tumor-targeted distribution of drug molecules, improve the therapeutic effect, and reduce the toxic and side effects on normal tissues and cells after functional modification. Patient-derived xenografts (PDX) models can be established by transplanting patient-derived cancer cells or small tumor tissue into immunodeficient mice directly. Compared with the tumor cell line model, this model can preserve the key features of the primary tumor such as histomorphology, heterogeneity, and genetic abnormalities, and keep them stable between generations. PDX models are widely used in drug evaluation, target discovery and biomarker development, especially providing a reliable research platform for the diagnosis and treatment evaluation of nano-delivery systems. This review summarizes the application of several common cancer PDX models in the evaluation of nano-delivery systems, in order to provide references for researchers to perform related research.

Research progress of intelligent reversible drug delivery system / 药学学报

In the research on cancer theranostics, most environment-sensitive drug delivery systems can only achieve unidirectional and irreversible responsive changes under pathological conditions, thereby improving the targeting effect and drug release performance of the delivery system. However, such irreversible changes pose potential safety hazards when the dynamically distributed delivery system returns to the blood circulation or transports to the normal physiological environment. Intelligent reversible drug delivery systems can respond to normal physiological and pathological microenvironments to achieve bidirectional and reversible structural changes. This feature will help to precisely control the drug release of the delivery system, prolong the blood circulation time, improve the targeting efficiency, and avoid the potential safety hazards of the irreversible drug delivery system. In this review, we describe the research progress of intelligent reversible drug delivery system from two main aspects: controlled drug release and prolonged blood circulation time/enhanced cellular internalization of drug.

Design, synthesis and antidiabetic activity studies of purine derivatives / 药学学报

Based our previous work, twelve purine derivatives were designed and synthesized as dual modulators of GPR119 and DPP-4by conjugating the GPR119 activating and DPP-4 inhibiting fragments with the position 6 and 9 of purine core via an approach of merged pharmacophores. Compound 11, bearing 2-fluoro-4-methylsulphonyl anilide and cyanopyrrolidine moieties, exhibited the most potent GPR119 agonistic activities (EC50 = 0.33 μmol·L-1, IA = 71.1%) and DPP-4 inhibitory (58.4% inhibition at 10 μmol·L-1, 21.2% inhibition at 1 μmol·L-1) activities in the in vitro antidiabetic study. Subsequently, we performed studies on structure activity relationships and molecular docking to guide the further drug design.

Effect of Huazhuo Jiedu Shugan Prescription on Learning， Memory， and PI3K/Akt/GSK-3β Signaling Pathway in Epileptic Rats / 中国实验方剂学杂志

Objective:To observe the effects of Huazhuo Jiedu Shugan Prescription （HZJDSG） on learning， memory， and the expression of phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt）/glycogen synthase kinase-3<italic>β</italic> （GSK-3<italic>β</italic>） pathway-related proteins in epileptic rats， and to explore its possible mechanism. Method:Forty-eight SPF male SD rats were randomly divided into a normal group， a model group， a sodium valproate （0.19 g·kg^-1） group， and low- （2.7 g·kg^-1）， medium- （5.4 g·kg^-1）， and high-dose （10.8 g·kg^-1） HZJDSG groups， with eight rats in each group. The normal group received 0.9% sodium chloride solution （0.035 g·kg^-1） by intraperitoneal injection， and the other five groups received pentetrazol （PTZ） at the same dose to induce a chronic epilepsy model for a total of 14 times. The drug groups received corresponding drugs and the normal group and the model group received 0.9% sodium chloride solution at the same volume once a day for 28 days. During the drug intervention period， epilepsy was maintained in each modeling group by intraperitoneal injection of PTZ on day 7， 14， 21， and 28. The behavioral changes of rats were observed by Morris water maze and the pathomorphological changes of rat hippocampal neurons by hematoxylin-eosin （HE） staining. The protein expression of phosphorylation Akt（p-Akt）and p-GSK-3<italic>β</italic> was detected by immunohistochemistry and the protein expression of PI3K， Akt， p-Akt， GSK-3<italic>β</italic>， and p-GSK-3<italic>β</italic> by Western blot. Result:Compared with the normal group， the model group showed prolonged platform finding time （<italic>P</italic><0.01）， reduced number of platform crossings （<italic>P</italic><0.01）， structural damage of neurons in the CA1 region of the hippocampus， down-regulated protein expression of p-Akt and p-GSK-3<italic>β </italic>in the CA1 region of the hippocampus （<italic>P</italic><0.05）， and reduced relative expression of PI3K， p-Akt， and p-GSK-3<italic>β</italic> in the hippocampus （<italic>P</italic><0.01）. Compared with the model group， the sodium valproate group and the HZJDSG groups showed shortened platform finding time （<italic>P</italic><0.01） and improved neuronal structure in the CA1 region of the hippocampus， while the sodium valproate group and the high- and medium-dose HZJDSG groups exhibited increased number of platform crossings （<italic>P</italic><0.01）， up-regulated protein expression of p-Akt and p-GSK-3<italic>β</italic> in the CA1 region of the hippocampus （<italic>P</italic><0.05）， and elevated relative expression of PI3K， p-Akt， and p-GSK-3<italic>β</italic> （<italic>P</italic><0.01）. Conclusion:HZJDSG can improve the learning and memory of epileptic rats， and its antiepileptic effect may be achieved by the activation of PI3K/Akt/GSK-3<italic>β</italic> pathway-related proteins.

Biomechanical study on femoroplasty-augmentation for prevention of osteoporotic hip fractures / 中国骨伤

OBJECTIVE@#To explore whether femoral plasty can improve the fracture resistance of osteoporotic femoral specimens and prevent hip fracture, and to compare the difference of mechanical strength changes between two different femoral plasty methods in osteoporotic femoral specimens, so as to determine the best strengthening area of the plasty.@*METHODS@#Eighteen pairs of fresh osteoporotic femur specimens were collected and divided into two groups, A and B, 9 pairs in each group. Nine fresh osteoporotic femur specimens in each group were randomly selected for enhancement, and the corresponding contralateral specimens were used as control group. In group A1, the enhancement areas were femoral head, femoral neck, femoral trochanter and subtrochantericregion. And in group B1, the enhancement areas were femoral head, femoral neck and femoral trochanter region. The amount of cement injected into the femoral neck was recorded and the surface temperature of the femoral neck was measured. All specimens were biomechanically tested under simulated falls. Load-displacement curves, final loads were recorded. The final energy and stiffness of specimens were calculated. The biomechanical differences between the specimens of the enhancement group and those of the corresponding control group were compared, and the mechanical changes of the specimens by two different enhancement methods were compared.@*RESULTS@#Compared with the control group, the ultimate load and energy of the specimens in the enhanced group increased significantly, but the stiffness did not change significantly. There was no significant difference in final load and energy between specimens strengthened by two different methods.@*CONCLUSION@#Femoral plasty has the advantages of minimally invasive, simple operationand remarkable effect. It can be used as a new method to prevent osteoporotic hip fracture.

Characterization of Vibrio cholerae isolates from 1976 to 2013 in Shandong Province, China

Abstract Cholera continues to be a serious public health issue in developing countries. We analyzed the epidemiological data of cholera from 1976 to 2013 in Shandong Province, an eastern coastal area of China. A total of 250 Vibrio cholerae isolates were selected for PCR analysis of virulence genes and pulsed-field gel electrophoresis (PFGE). The analysis of the virulence genes showed that the positive rates for tcpA and tcpI were the highest among strains from the southwest region, which had the highest incidence rate of cholera. Low positive rates for tcpA, tcpI and ctxAB among isolates from after 2000 may be an influencing factor contributing to the contemporary decline in cholera incidence rates. Spatiotemporal serotype shifts (Ogawa, Inaba, Ogawa, Inaba and O139) generally correlated with the variations in the PFGE patterns (PIV, PIIIc, PIa, PIIIb, PIIIa, PIb, and PII). O1 strains from different years or regions also had similar PFGE patterns, while O139 strains exclusively formed one cluster and differed from all other O1 strains. These data indicate that V. cholerae isolates in Shandong Province have continually undergone spatiotemporal changes. The serotype switching between Ogawa and Inaba originated from indigenous strains, while the emergence of serogroup O139 appeared to be unrelated to endemic V. cholerae O1 strains.

Chemical studieson hydrophilic constituents of toad skin / 中国中药杂志

Fifteen compounds were isolated from the toad skin by a combination of various chromatographic methods including macroporous resin, silica gel, ODS and semi-preparative HPLC. Their structures were identified as 4,5-dimethyl-1,3,4,5-tetrahydropyrrolo[4,3,2-de]quinolin-6-ol(1), serotonin(2), N-methyl serotonin(3), O-methyl bufotenine(4), 1,2,3,4-tetrahydro-6-hydroxy-β-carboline(5), O-methylserotonin(6), glycinebetaine(7), caffeine(8), bufotenine(9), shepherdine(10), tryptophan(11), (5-hydroxy-1H-indol-3-yl)acetic acid(12), 5-hydroxy tryptophol(13), 2-methyl-6-hydroxy-1,2,3,4-tetrahydro-β-carboline(14), bufothionine(15). Among them, compound 1 was a new compound,compound 5 was a new natural product. Compounds 4-8 and 10-14 were separated from toad skin for the first time.

Immunophenotypic Analysis of Acute Promyelocytic Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the immunophenotype of leukemia promyelocytes (LP) in bone marrow of patients with acute promyelocytic leukemia (APL) and to explore their characteristics and significance.</p><p><b>METHODS</b>The immunophenotypes of leukemia cells in 43 patients with APL were analyzed by means of 4 color immunophenotypes; the cell population in which CD45 strength localized at 10(2) and the SSC strength locatized at 10(2) was defined as R3, the cell population in which CD45 strength localized at 10(3) and the SSC strength localized at 10(2) was defined as R5, moreover the ratio of positive cells >80% was defined as strong positive expression, the ratio of positive cells between 20%-80% was difined as weak positive expression, the ratio of positive cells <20% was difined as negative by gating method of CD45/SSC.</p><p><b>RESULTS</b>There was a abnormal cell population (R3) in 79.07% cases; the immunophenotypes of R3 was cheracteried by high SSC, weaker expression of CD45, the rate of CD38, CD9 and CD13 all was 100%, moreover their bright expression (>80%) was 86.05%, 90.70% and 86.05%, respectively; the positive expression rate of CD33, CD117 and CD64 was 97.67%, 95.35% and 83.80% respectively, moreover thier bright expression was 84.04%, 69.77% and 30.23% respectively; the CD15 was weakly expressed in 39.53% cases, the CD34 and HLA-DR were weakly expression in 16.28% and 6.98% cases respectively. All the cases did not express CD116. There were 2 cell populations (R3 and R5) in 20.93% cases, the immunophenotypic features of R3 were cosistant with above mentioning, while the immunophenotypes of R5 were lower than those of R3 SSC; the fluorescence intensity of CD45 was higher, but lower than that in normal lymphycytes, the positive rate of CD9, CD13, MPO was 100%, moreover thier fluorescence intensity was high; they did not expressed CD123, CD25, CD22, CD4, CD64 and CD14. Thereby it can be concluded that the typical immunophenotypes is characterized by CD13(+) CD9(+) CD38(+) CD33(+) CD117(+) CD64(+) CD11b(-) CD34(-) HLA-DR(-) in APL. There was a special immunophenotype in the APL with basophilic granules. Conclusoin: APL has a characteristic immunophenotypic profile, whose typical immunophenotype is characterized by CD13(+) CD9(+) CD38(+) CD33(+) CD117(+) CD64(+) CD11b(-) CD34(-) HLA-DR(-). The special immunophenotype exists in the APL with basophilic granules. Flow cytometric immunophenotyping may be a useful for rapid recognition of APL and has significant for prognosis.</p>

Activatable cell-penetrating peptides: a potential activatable modality for diseases diagnosis and therapy / 药学学报

Cell-penetrating peptides are composed of positively-charged amino acids that can mediate molecules or nano-carriers across cell membranes. However, most of the known cell-penetrating peptides have no cell-or tissue-specificity, with affinity to almost all types of cells in internalization. The non-specificity of cell-penetrating peptides is a significant obstacle in the application to targeted delivery of imaging probes and therapeutic agents. Accordingly, many studies focused on selective switching of systemically-delivered inert cell-penetrating peptides into active forms in diseased tissues. Tsien groups introduced the concept of activatable cell-penetrating peptides in 2004. Subsequently, a growing number of similar delivery systems (molecular or nano-sized) have been documented, and the sensitive factors have included enzyme, lower pH, light and exogenous component. In this paper, we make an overview of the development of activatable delivery system in recent years.

Clinical characteristics and surgical treatment of ganglioneuroma in spine / 中国骨伤

<p><b>OBJECTIVE</b>To summarize the clinical manifestation and diagnosis of ganglioneuroma in spine and investigate the clinical effect of surgical treatment.</p><p><b>METHODS</b>The clinical data of 6 patients underwent a surgery for ganglioneuroma in spine from January 2008 to January 2015 were retrospectively analyzed. There were 4 males and 2 females, aged from 2 to 63 years old with an average of 34.6 years. The courses of disease were from 3 days to 17 years. Five patients complicated with superficial hypesthesia in correlative level of tumor, and the muscle strength under tumor plane had decreased at different levels, with the strength of grade II-IV. Two cases complicated with hypermyotonia and positive bilateral Hoffmann's and Babinski sign. Five cases were sporadic lesion in correlative spinal canal and one case complicated with the giant occupying lesion in thoracic cavity.</p><p><b>RESULTS</b>Six operations had been performed including 5 en bloc and 1 subtotal resection. Postoperative pathological results showed tumor cells scattered or fasciculated inserted into Schwann cells in the stroma. In 2 patients complicated with radiculalgia before operation, 1 case was relieved and 1 was invariant after operation. All 4 patients with preoperative dyscinesia in the limbs obtained improvement after operation. All the patients were followed up from 0.3 to 6.8 years with an average of 2.5 years. At the final follow-up, according to ASIA grade, 5 cases were good and 1 case was invariant. During the follow-up, only 1 patient experienced chemoradiation because of merging ganglioneuroblastoma and receiving subtotal resection. No recurrence in other 5 cases.</p><p><b>CONCLUSION</b>Ganglioneuroma is a benign and rare tumors in spine. Clinically, radicular pain and sensory-motor disorders are the main manifestations. Its diagnosis depends on pathological examination. Prognosis of surgical treatment is good.</p>

Phosphorylation of eukaryotic initiation factor 2-alpha inhibits cisplatin-mediated apoptosis of hepatocellular carcinoma cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate whether the phosphorylation (functionally inhibitive) of eukaryotic initiation factor 2-alpha (eIF2-a) affects the molecular mechanism of cisplatin-induced cellular apoptosis in human hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The human HCC cultured cell lines SMMC-7221 and HepG2 were treated with cisplatin alone (controls; 24 h) or in combination with pre-transfection of a dominant-negative eIF2-a mutant (eIF2aS51A) or pre-exposure to an eIF2-a-specific phosphatase inhibitor (salubrinal) to decrease or increase the phosphorylation level, respectively. Changes in expression of apoptosis markers were quantitatively and qualitatively assessed by flow cytometry and western blot analysis. The significance of differences among groups was assessed by analysis of variance testing and of differences between groups was assessed by t-test.</p><p><b>RESULTS</b>Cisplatin treatment induced the appropriate functional-inhibitive phosphorylation of eIF2-a on serine 51. Cisplatin treatment (10 mg/ml) induced significant apoptosis in the eIF2aS51A pre-transfected SMMC-7721 (control: 21.7 +/- 1.5% vs. 50.7 +/- 2.1%, t = 19.454, P less than 0.05) and HepG2 (21.0 +/- 1.0% vs. 57.3 +/- 2.1%, t = 27.250, P less than 0.05). Salubrinal pre-treatment significantly inhibited the cisplatin (15 mg/ml)-induced apoptosis in SMMC-7721 (control: 50.3 +/- 2.5% vs. 16.3 +/- 2.1%, t = 18.031, P less than 0.05) and HepG2 (42.0 +/- 2.6% vs. 12.0 +/- 2.0%, t = 15.667, P less than 0.05).</p><p><b>CONCLUSION</b>Phosphorylation of eIF2-a may act to inhibit cisplatin-induced apoptosis of HCC.</p>

Treatment strategy of double disruption of the superior shoulder suspensor complex (SSSC) / 中国骨伤

<p><b>OBJECTIVE</b>To discuss efficacy of the surgical treatment strategy of double disruption of the superior shoulder suspensor complex (SSSC).</p><p><b>METHODS</b>The data of 15 patients with double disruption of the SSSC were retrospectively analyzed from January 2008 to December 2009. There were 11 males and 4 females, with an average age of 45.1 years (ranged, 19 to 60). Of them, 8 patients were treated with surgery and 7 patients with conservative treatment. The short-term effectiveness was evaluated according to Constant-Murley scoring system.</p><p><b>RESULTS</b>All patients were followed up for 7 to 24 months with an average of 14.4 months. All fractures healed with a mean time of 12.3 weeks (ranged, 9 to 12). At final follow-up, the Constant-Murley scores with patients of surgical treatment was (92.37 +/- 1.99), and patients of conservative treatment was (55.52 +/- 1.29).</p><p><b>CONCLUSION</b>Surgical treatment can restore the integrality of SSSC, in favour of stability between upper limb and trunk.</p>

Treatment of tibiofibular diastasis without ankle fracture with endobutton device / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the clinical effects of Endobutton device in the treatment of tibiofibular diastasis without ankle fracture.</p><p><b>METHODS</b>From January 2009 to January 2011, a total of 8 patients with tibiofibular diastasis without ankle fracture were treated with Endobutton device. There were 6 males and 2 females with an average age of 34 years (ranged, 25 to 44 years). All the patients with ankle injured history and ankle pain, swelling, ecchy-mosis were diagnosed by radiology and then operated with Endobutton device. The clinical effects were evaluated according to Baird-Jackson scoring system and radiological evaluated parameters comprised of tibiofibular overlap,total clear space and medial clear space.</p><p><b>RESULTS</b>All the patients were followed up, and the duration ranged from 6 to 24 months,with an average of 12 months. Radiographic results were detailed as follows: tibiofibular overlap averaged (3.83 +/- 0.37) mm in preoperative and (7.46 +/- 0.14) mm in postoperative; mean total clear space (7.90 +/- 0.22) mm in preoperative and (3.39 +/- 0.07) mm in postoperative; medial clear space averaged (5.08 +/- 0.34) mm in preoperative and (3.16 +/- 0.07) mm in postoperative. There was significant difference above data between preoperative and postoperative one. The lastest Baird-Jackson score results: 6 cases obtained an excellent result, 1 good and 1 fair. The main score was (94.63 +/- 3.66).</p><p><b>CONCLUSION</b>Endobutton device for the treatment of tibiofibular diastasis without ankle fracture has advantages such as simple and minimally invasive, no need of second operation for implant removal, recover the ankle function better and less complications, which should be popularized and applied to clinical widely.</p>

Current situation and progression of induced pluripotent stem cells in treating spinal cord injury / 中国骨伤

Spinal cord injury is a difficult medical problem and current therapeutic methods could not obtain satisfactory results. Recent 20 years, stem cell technology developed rapidly, embryonic stem cells and adult stem cells were used for treating neurological disease and nerve injury of animal models and the clinical results were confirmed. It provided a new prospect for the treatment of nerve injury at the cellular level. However,due to technical and ethical problems, it is difficult to obtain the appropriate cells that can be applied to the human being. Recently, induced pluripotent stem cells were developed as a new method for the treatment of spinal cord injuries by the autologous transplantation. Starting from this work, the purpose of this review is to assess the differentiate ability of induced pluripotent stem cells into neurocyte and review the latest developments in this area.

The role of mir-221/222 in inhibiting endoplasmic reticulum stress-induced human hepatocarcinoma cell apoptosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the role of miR-221/222 in inhibiting endoplasmic reticulum stress-induced human hepatocarcinoma cells apoptosis.</p><p><b>METHOD</b>miR-221/222 mimics and inhibitors were used to mimic or block the function of endogenous miR-221/222 respectively. Western blot and flow cytometry were used to test the effects of miR-221/222 on cell cycle and apoptosis under endoplasmic reticulum stress in human hepatocellular carcinoma cells.</p><p><b>RESULTS</b>Endoplasmic reticulum stress resulted in miR-221/222 down-regulation in human hepatocellular carcinoma cells. miR-221/222 mimics and inhibitors inhibited and promoted respectively endoplasmic reticulum stress-mediated p27Kip1 induction. Moreover, p27Kip1 suppression not only resulted in reduction in the fraction of G1 phase cells, but also promoted the endoplasmic reticulum stress-mediated apoptosis in human hepatocellular carcinoma cells.</p><p><b>CONCLUSION</b>miR-221/222 were downregulated by endoplasmic reticulum stress in human hepatocellular carcinoma cells, which subsequently protected human hepatocellular carcinoma cells against endoplasmic reticulum stress-induced apoptosis through p27Kip1 regulation.</p>

Beta-arrestin2 stimulates interleukin-17 production and expression of CD4+ T lymphocytes in a murine asthma model

Allergic asthma is a complex and chronic inflammatory airway disease. Interleukin-17 is a pro-inflammatory cytokine which plays critical role in the pathogenesis of allergic asthma. It has been reported that beta-arrestin2 regulated the development of allergic asthma at a proximal step in the inflammatory cascade. In this study, the influence of beta-arrestin2 on Interleukin-17 production and expression of CD4+ T lymphocytes in a murine asthma model was investigated. Splenic CD4+ T lymphocytes from wild-type mice and those from a murine asthma model were purified. CD4+ T lymphocytes from a murine asthma model were transfected with siRNAs targeting the beta-arrestin2 or were pretreated with the ERK1/2 inhibitor, PD98059. After stimulation, the protein expression of beta-arrestin2 phosphorylated-ERK1/2 and IL-17 were detection by Western blot; the mRNA expression of IL-17 were detected by real-time PCR; the accumulation of IL-17 in supernatants were detected by ELISA. We found that beta-arrestin2 phosphorylated-ERK1/2 and IL-17 expression in CD4+ T lymphocytes from a murine asthma model was increased compared with those from wildtype mice[p<0.01]. Treatment of CD4+ T lymphocytes with siRNAs targeting the beta-arrestin2 down-regulated phosphorylated- ERK 1/2 and IL-17 expression [p < 0.01]. PD98059 decreased IL-17 production and expression in CD4+ T lymphocytes in a murine asthma model [p < 0.05]. We conclude that beta-arrestin2 stimulated IL-17 production and expression of CD4+ T lymphocytes in a murine asthma model. The effect was partly mediated by ERK 1/2 activation. Targeting beta-arrestin2 biological activity could be a valid therapeutic approach for the treatment of allergic asthma

Inhibitory effect of interference hTERT and TRF2 gene on the growth of breast cancer MCF-7 cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the effect of combined gene therapy with interference hTERT and TRF2 gene on the treatment of breast cancer.</p><p><b>METHODS</b>Recombinant adenovirus rAd-hTERT and rAd-TRF2 expressing siRNA-hTERT and siRNA-TRF2 was constructed, and the vectors were transfected into MCF-7 cells. Than the expressions of hTERT and TRF2 proteins were detected by Western blot, the inhibition of MCF-7 cell proliferation by MTT colorimetry, and the changes of MCF-7 cell cycle by flow cytometry and the colony forming ability of MCF-7 cells by clone form test.</p><p><b>RESULTS</b>At 48 h after transfection, the relative expression amounts of hTERT protein of the PBS control group, rAd-blank group, rAd-HK control group, rAd-hTERT group, rAd-TRF2 group and rAd-hTERT and rAd-TRF2 group were 1.00, 0.94 +/- 0.02, 0.95 +/- 0.04, 0.18 +/- 0.04, 0.95 +/- 0.01 and 0.18 +/- 0.04, respectively. The relative expression amounts of TRF2 protein were 1.00, 1.01 +/- 0.08, 0.96 +/- 0.02, 0.95 +/- 0.08, 0.22 +/- 0.01 and 0.26 +/- 0.02, respectively. After transfection of rAd-hTERT or rAd-TRF2 into MCF-7 cells separately, the inhibition rate of cell proliferation was only 54.6% and 48.4%, there was 8.9% +/- 1.2% or 9.2% +/- 2.3% of MCF-7 cells into M phase, 66.4% +/- 1.5% or 64.6% +/- 1.9% of MCF-7 cells was arrested at G(0)/G(1) phase, and the cell colony forming ability was decreased significantly (cell colony number from 100 in PBS control group down to 41.3 +/- 5.1 and 43.7 +/- 6.4). But after transfection by rAd-hTERT and rAd-TRF2 simultaneously, the inhibition rate of cell proliferation was about 82.1%, and M phase cells was significantly reduced to 4.4% +/- 1.2%. Large numbers of cells were arrested at G(0)/G(1) phase (81.4% +/- 1.3%), and the cell colony forming ability was more significantly decreased (cell colony number there were only 29.2 +/- 3.9).</p><p><b>CONCLUSION</b>More effective effect of tumor gene therapy can be achieved by combination of interference hTERT and TRF2 genes as compared with interference by either of the single gene alone.</p>

Cross-talk between PI3K/Akt and MEK/ERK pathways regulates human hepatocellular carcinoma cell cycle progression under endoplasmic reticulum stress / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the cross-talk between the PI3K/Akt and MEK/ERK pathways and its role in cell cycle regulation under endoplasmic reticulum stress in human hepatocellular carcinoma cells.</p><p><b>METHODS</b>PI3K inhibitor LY294002 and MEK inhibitor U0126 were used to block the PI3K/Akt and MEK/ERK pathways respectively, and constitutively activated Akt mutant construct was used to activate the PI3K/Akt pathway. Western blot was used to study the potential cross-talk between the PI3K/Akt and MEK/ERK pathways under endoplasmic reticulum stress in human hepatocellular carcinoma cells. the role of the cross-talk between the PI3K/Akt and MEK/ERK pathways in cell cycle regulation was investigated by using propidium iodide staining.</p><p><b>RESULTS</b>LY294002 not only blocked Akt activation efficiently but also increased ERK phosphorylation markedly under endoplasmic reticulum stress in SMMC-7721 and Hep3B cells. Furthermore, myr-Akt inhibited endoplasmic reticulum stress-mediated ERK phosphorylation. In contrast, MEK inhibitor U0126 had no effect on endoplasmic reticulum stress-induced Akt activation. It is notable that both myr-Akt overexpression and MEK inhibitor U0126 inhibited endoplasmic reticulum stress-induced G0/G1 phase arrest in SMMC-7721 cells.</p><p><b>CONCLUSION</b>Endoplasmic reticulum stress-induced Akt activation is mediated through PI3K and the PI3K/Akt pathway inactivation is involved in increased ERK activity in human hepatocellular carcinoma cells. The cross-talk between the PI3K/Akt and MEK/ERK cascades plays an important role in endoplasmic reticulum stress-induced human hepatocellular carcinoma cell cycle arrest.</p>

Application of elastic intramedullary nail in treating bilateral femoral shaft fractures in children / 中国骨伤

<p><b>OBJECTIVE</b>To explore the curative effects and complications of elastic intramedullary nail in treating children's bilateral femoral shaft fractures.</p><p><b>METHODS</b>Form February 2005 to March 2008, 7 patients with bilateral femoral shaft fractures were treated by closed reduction and internal fixation with elastic intramedullary nail. There were 5 males and 2 females. The age ranged from 3 to 13 years with the mean of 8.3 years. Six injuries caused by road accident and 1 injury caused by fall from high. Two cases associated with pulmonary contusion, 3 cases brain injuries, 1 case fracture of calcaneus and 1 case bladder injuries. All the cases were closed fractures without nerve and blood vessel injury. A cast external fixation had been used after operation for a month in two cases.</p><p><b>RESULTS</b>All the patients were followed up for 21-37 months with an average of 30.3 months. No infecton of incisional wound, displacement fracture, internal fixation fail, delayedunion and malunion were found. All fracture obtained healing for 7-12 weeks with an average of 8.7 weeks. Inequality of lower limb was found in 1 case (length differences was 5 mm). According to Flynn scoring,all fractures were excellent.</p><p><b>CONCLUSION</b>Treatment of femoral shaft fractures in children with elastic intramedullary nail according with biological principle. The method has little trauma, less complication, outstanding effect and it is a good way to treat bilateral femoral shaft fractures result from high-energy injuries.</p>