In this paper, in vitro anti-influenza virus activities of sulfated polysaccharide fractions from Gracilaria lemaneiformis were investigated. Cytotoxicities and antiviral activities of Gracilaria lemaneiformis polysaccharides (PGL), Gracilaria lemaneiformis polysaccharide fraction-1 (GL-1), Gracilaria lemaneiformis polysaccharide fraction-2 (GL-2) and Gracilaria lemaneiformis polysaccharide fraction-3 (GL-3) were studied by the Methyl thiazolyl tetrazolium (MTT) method, and the inhibitory effect against Human influenza virus H1-364 induced cytopathic effect (CPE) on MDCK cells were observed by the CPE method. In addition, the antiviral mechanism of PGL was explored by Plaque forming unit (PFU), MTT and CPE methods. The results showed: i) Cytotoxicities were not significantly revealed, and H1-364 induced CPE was also reduced treated with sulfated polysaccharide fractions from Gracilaria lemaneiformis; ii) Antiviral activities were associated with the mass percentage content of sulfate groups in polysaccharide fractions, which was about 13%, in polysaccharides (PGL and GL-2) both of which exhibited higher antiviral activity; iii) A potential antiviral mechanism to explain these observations is that viral adsorption and replication on host cells were inhibited by sulfated polysaccharides from Gracilaria lemaneiformis. In conclusion, Anti-influenza virus activities of sulfated polysaccharide fractions from Gracilaria lemaneiformis were revealed, and the antiviral activities were associated with content of sulfate groups in polysaccharide fractions.

Objective To detect the selective inhibitory effects of irradiation plus adenovirusmediated horseradish peroxidase ( HRP)/indole-3-acetic acid (IAA) suicide gene system using tumorspecific and radio-inducible chimeric promoter on human hepatocellular carcinoma subcutaneously xenografted in nude mouse.MethodsRecombinant replicated-deficient adenovirus vector containing HRP gene and chimeric human telomerase reverse transcriptase (hTERT) promoter carrying 6 radioinducible CArG elements was constructed.A human subcutaneous transplanting hepatocellular carcinoma (MHCC97 cell line) model was treated with -γ-ray irradiation plus intra-tumor injections of adenoviral vector and intra-peritoneal injections of prodrug IAA.The change of tumor volume and tumor growth inhibiting rate,the survival time of nude mice,as well as histopathology of xenograft tumor and normal tissues were evaluated.Results Thirty one days after the treatment,the relative tumor volumes in the negative,adenovirus therapy,irradiation,and combination groups were 49.23 ± 4.55,27.71 :± 7.74,28.53 + 10.48 and 11.58 ± 3.23,respectively.There was a significantly statistical difference among them (F = 16.288,P ＜0.01 ).The inhibition effect in the combination group was strongest as compared with that in other groups,and its inhibition ratio was 76.5%.The survival period extended to 43 d in the combination group,which showed a significantly difference with that in the control group(x2 = 18.307 ,P ＜0.01 ).The area of tumors necrosis in the combination group was larger than that in the other groups,and the normal tissues showed no treatment-related toxic effect in all groups.However,multiple hepatocellular carcinoma metastases were observed in the liver in the control group,there were a few metastases in the monotherapy groups and no metastasis in the combination group.Conclusions Adenovirus-mediated suicide gene therapy plus radiotherapy dramatically could inhibit tumor growth and prolong median survival time.It might provide a promising therapeutic modality for hepatocellular carcinoma therapy.

Objective To evaluate the value of transvaginal ultrasound in diagnosing intrauterine adhesions.Methods Transvaginal ultrasound was performed in 136 patients with suspicious intrauterine adhesions and compared with hysteroscopy correspondingly .The ultrasonographic features of intrauterine adhesions on transvaginal ultrasound were summarized .Results One hundred and twenty one cases (89.0%, 121/136 ) of intrauterine adhesions were verified by hysteroscopy .The hysteroscopic findings included:(1) Forty seven cases(38.9%,47/121) were minimal intrauterine adhesions , 46 cases(38.0%, 46/121) were moderate intrauterine adhesions , and 28 cases (23.1%,28/121) were severe intrauterine adhesions.(2) Sixty one cases(50.4%,61/121) were central intrauterine adhesions , 24 cases(19.8%, 24/121) were marginal intrauterine adhesions , and 36 cases (29.8%, 36/121) were mixed type of intrauterine adhesions.The transvaginal ultrasound findings included:(1)Nineteen cases(40.4%,19/47) were minimal intrauterine adhesions ,33 cases(71.7%,33/46)were moderate intrauterine adhesions ,and 23 cases(82.1%,23/28) were severe intrauterine adhesions .(2) Thirty nine cases (63.9%,39/61) were central intrauterine adhesions ,9 cases(37.5%,9/24) were marginal intrauterine adhesions ,and 27 cases (75.0%, 27/36 ) were mixed type of intrauterine adhesions .By transvaginal ultrasound, seventy-five (62.0%,75/121) cases of intrauterine adhesions were correctly diagnosed , whereas 46 cases (38.0%, 46/121) were missed.And 3 cases ( 3.8%, 3/78 ) were misdiagnosed as intrauterine adhesions on transvaginal ultrasound,including one endometrial polyp ,one thin endometrium and one septate uterus .The sensitivity, specificity and accuracy of transvaginal ultrasound in diagnosing intrauterine adhesions were 62.0%(75/121), 80.0%(12/15) and 64.0%(87/136) respectively.There were significant statistical differences in diagnosing different degrees of intrauterine adhesions ( χ2 =15.956,P=0.000) and different parts of intrauterine adhesions( χ2 =8.792,P=0.012) by transvaginal ultrasound.Conclusions Transvaginal ultrasound is an effective, easy to perform and noninvasive technique in screening and diagnosing intrauterine adhesions.Transvaginal ultrasound is an effective way in diagnosing intrauterine adhesions showing a noninvasive and simpler way than hysteroscopy .Transvaginal ultrasound is of great value in screening and diagnosing intrauterine adhesions .

Analogous formulae (AF) refer to a set of traditional Chinese medicine (TCM) formulae sharing similar herbs and/or indications. Dissecting functional chemome of analogous formulae could enhance the understanding of the intrinsic nature of TCM. In this study, taking 5 Xiaoqinglong decoction analogous formulae (XQL AF) including Xiaoqinglong decoction, Mahuang Xingren Shigao Gancao decoction, Mahuang Fuzi decoction, Houpu Mahuang decoction and Daqinglong decoction as example, we systematically investigated the relationship between compounds and indications using network formulaology approach. The functional chemome of XQL AF were revealed by network analysis and molecular docking. This successful application in XQL AF suggests network formulaology could be a useful tool for AF-related research and therefore provide a new way to discover the scientific foundation of Zhang Zhongjing's herbal formulae.
Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; analysis ; Medicine, Chinese Traditional ; Molecular Docking Simulation

Objective To develop a convenient reverse transcription PCR(RT-PCR)method for identifying genotypes of human metapneumovirus(hMPV)from clinical samples.Methods According to the gene sequences of hMPV G with different genotypes,the A and B genotype specific primers were designed.A diplex RT-PCR was applied to identify different genotypes according to the molecular weight of PCR products in agarose gel.37 clinical samples were detected through this method.Results It was convenient to distinguish different genotypes of hMPV(383 bp for A and 284 bp for B)by the diplex RTPCR,and there was no non-specific amplification for common respiratory viruses.so it meant that the specificity of primers was good.The results of genotyping 37 clinical samples showed that 20 samples were identified as genotype A by both sequence analysis of M gene and diplex RT-PCR,whereas 17 samples were identified as genotype B by sequence analysis of M gene.but in these 17 samples 14 samples were identified as genotype B by the diplex RT-PCR and remaining 3 samples could not be genotyped because there was no PCR product after amplification.The consistency rate for these two methods Was 91.9%[(20+14)/37].Conclusion The method of diplex RT-PCR Was developed successfully and can be used for identify genotypes of hMPV.

Objective To construct a detailed, 3-dimensional, anatomically accurate finite element (FE) model of lumbar L4-L5 segment from CT data with a new kind of computer aided design (CAD) method. Methods A modified "no-seed region segmentation" was done to extract the interest region in the CT scan images and produce a binary image. "Best cross-section planes" accounting for the preferential direction dictated by lumbar spine were placed on the initial iso-surface model, forming a "non-regular piecewise subspace". This subspace and the embedded iso-surface mode were transformed by local affine transforms to a "regular subspace", in which a surface mesh of high quality was generated quickly. Finally a reverse transform procedure was employed to recover the shape feature of the lumbar surface mesh of lumbar L4-L5 in the original 3-dimensional space, which was then imported into ANSYS for the 3-dimensional FE mesh construction. Results All complicated anatomical features of the L4-L5 segment were explicitly represented in the unprecedented finite element model. The predicted results for compression, flexion and extension correlated well with experimental data under similar loading configurations. Conclusion The presented CAD method containing advanced algorithm implements fast and accurate simulation of such complicated geometry with fine mesh representation for lumbar FE analysis.

Objective To observe the effect of very early intervention with cerebellar fastigial nucleus electrical stimulation and neurodevelopmental treatment on infants of central coordination disturbance(CCD).Methods 86 infants(0～6 months)with central coordination disturbance were divided into intervention group and control group.The intervention group was treated with neurodevelopmental treatment,cerebellar fastigial nucleus(FN)electrical stimulation and family interference.The control group was treated with neurodevelopmental treatment and family interference.The effect of infant was assessment with Gross Motor Function Measure(GMFM)and Gesell Development Test after 3 months treatment.Results The total improved incidence of the intervention group was 95.6%,which was significantly higher than that of the control group(P<0.01).The difference of scores in social,adaptive and motor area improved in the intervention group compared with that of control group(P<0.05).Conclusion Very early intervention with cerebellar fastigial nucleus electrical stimulation can facilitate the development of gross motor,social,adaptive capability of infants with CCD treated with neurodevelopmental therapy.

OBJECTIVETo study the influences of endocannabinoid-anandamide (AEA) on the proliferation and apoptosis of the colorectal cancer cell line (CaCo-2) and to elucidate the effects of CB1 and lipid rafts, and to further elucidate the molecular mechanism and the effect of AEA on the generation and development of colorectal cancer.

METHODSHuman colorectal cancer cell line CaCo-2 was cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum in 5% CO(2) atmosphere at 37°C. CaCo-2 cells were divided into different groups and treated with different concentrations of AEA, AEA + SR141716A, AEA + AM630 and AEA + methyl-β-cyclodextrin (MCD). MTT assay was used to determine the effects of AEA, its putative CB1, CB2 receptor antagonists (SR141716A and AM630) and MCD on the proliferation of CaCo-2 cells. Annexin V-PE/7AAD binding assay was used to detect apoptosis in the CaCo-2 cells. Western-blot was applied to check the expressions of CB1, CB2, p-AKT and caspase-3 proteins in different groups of CaCo-2 cells.

RESULTSAEA inhibited the proliferation of CaCo-2 cells in a concentration-dependent manner and the effect could be antagonized by SR141716A and MCD. The inhibiting rates were (21.52 ± 0.45)%, (42.16 ± 0.21)%, (73.64 ± 0.73)% and (83.28 ± 0.71)%, respectively, at different concentrations of AEA (5, 10, 20 and 40 µmol/L). The three groups (20 µmol/L AEA, 20 µmol/L AEA + 10 µmol/L SR141716A and 20 µmol/L AEA + 1 mmol/L MCD) showed different inhibiting rates [(73.64 ± 0.73)%, (16.15 ± 0.75)% and (12.58 ± 0.63)%], respectively. Annexin V-PE/7AAD binding assay showed that AEA induced apoptosis in the CaCo-2 cells and MCD could antagonize this effect. The apoptosis rates of the three groups (control, 20 µmol/L AEA and 20 µmol/L AEA + 1 mmol/L MCD) were (2.95 ± 0.73)%, (39.61 ± 0.73)% and (14.10 ± 0.64)%, respectively. The expressions of CB1, CB2, p-AKT and Caspase-3 proteins were all observed in the CaCo-2 cells. AEA inhibited p-AKT protein expression and induced caspase-3 protein expression. The two actions were also antagonized by MCD.

CONCLUSIONSAEA can strongly suppress the proliferation of colorectal cancer CaCo-2 cells via the CB1 receptor and membrane cholesterol-LRs and induce apoptosis via lipid rafts. Anandamide plays a very important role in the carcinogenesis and development of colorectal cancer. MCD is a critical member in this system.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Arachidonic Acids ; antagonists & inhibitors ; pharmacology ; Caco-2 Cells ; Cannabinoid Receptor Modulators ; antagonists & inhibitors ; pharmacology ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Endocannabinoids ; Humans ; Indoles ; pharmacology ; Membrane Microdomains ; metabolism ; Piperidines ; pharmacology ; Polyunsaturated Alkamides ; antagonists & inhibitors ; pharmacology ; Proto-Oncogene Proteins c-akt ; metabolism ; Pyrazoles ; pharmacology ; Receptor, Cannabinoid, CB1 ; antagonists & inhibitors ; metabolism ; Receptor, Cannabinoid, CB2 ; antagonists & inhibitors ; metabolism ; beta-Cyclodextrins ; metabolism

OBJECTIVE: To design and manufacture a new type instrument and its matching tools for the proximal femur. METHODS: A new type instrument of subtrochanterical fracture-static 3-dimensional screw-plate system(STDSP)was designed and manufactured. The different types of accessory instruments which adapted to STDSP,the aiming guide, and the accessory tools were also designed and manufactured, and they were tested on bones. RESULTS: STDSP included 3 special lag screws and 1 anatomic plate. The lag screws distributed in femoral neck like "or"shape. There was 135 degree between the lag screws and femoral shaft. The shape of the plate was similar to the anatomic structure of the proximal thighbone. The posterior screw was stably connected to the plate. The plate was affixed crosswise to the lateral and anterior side by bicortical screws. The aiming guides and all kinds of accessory tools could help to install STDSP exactly and conveniently,which was verified on cadavers and X-ray photographs. CONCLUSION STDSP has the function of dynamic compressing in 3-dimensional space, and can match with the structure of the proximal femur preferably. STDSP may be a suitable instrument to the subtrochanteric fractures. The accessory apparatus of STDSP has simple configuration,and can let the STDSP be installed accurately, conveniently and rapidly.
Bone Plates ; Bone Screws ; Cadaver ; Equipment Design ; Fracture Fixation, Intramedullary ; instrumentation ; Hip Fractures ; surgery ; Humans

Neonatal chylothorax is a common cause of neonatal congenital pleural effusion and is often caused by the accumulation of chylous fluid in the thoracic cavity due to the rupture of the thoracic duct and its branched lymphatic vessels for a variety of reasons. Neonatal chylothorax caused by malignant tumors is extremely rare, and this is the first case of neonatal mediastinal neuroblastoma with chylothorax in China. The boy was found to have pleural effusion in the left thoracic cavity in the uterus, and experienced apnea at birth, as well as dyspnea and cyanosis as the main manifestations after birth. He was diagnosed with left chylothorax based on conventional biochemical analysis of pleural effusion. After the treatment including persistent chest drainage and symptomatic and supportive treatment, the drainage of the left thoracic cavity reached a volume of 90-180 mL per day. Neonatal refractory chylothorax was considered. Chest radiograph on day 13 after birth showed lesions in the upper left lung field, and contrast-enhanced plain CT scan of the chest suggested the possibility of posterior mediastinal neuroblastoma. The autopsy confirmed giant posterior mediastinal neuroblastoma (poorly differentiated), which involved the C7-T6 spinal canal and the nearby erector spinae, with a small amount of tumor tissue in the liver and both adrenal glands. Mediastinal tumor is considered the underlying cause of chylothorax in this case.
China ; Chylothorax ; Dyspnea ; Female ; Humans ; Infant, Newborn ; Male ; Pleural Effusion ; Uterus