Polysaccharides with multiple biological activities are usually considered as one of the major bioactive compounds in Chinese medicines (CMs). At present, the development of drug and functional foods related to polysaccharides have attracted a great deal of attention due to their great potential effects and diverse action mechanisms. However, quality control of polysaccharides is the bottleneck and a challenge due to their complexity and chemical diversity. Actually, the bioactivities of polysaccharides are closely related to their molecular structures. In order to ensure their safety and efficacy, the development of novel approaches based on the molecular structures for the improvement of quality control of polysaccharides is significantly important. Therefore, in this article, the relationship between biological activities and chemical structures, as well as the action mechanisms of polysaccharides from CMs were summarized first. Furthermore, saccharide mapping, a novel strategy for quality control of bioactive polysaccharides from CMs, was introduced and the application and perspectives were also discussed.
Carbohydrate Sequence ; Drugs, Chinese Herbal ; chemistry ; Molecular Sequence Data ; Polysaccharides ; chemistry ; Quality Control

Objective The prognosis of traumatic brain injury is closely associated with the apoptosis of neural cells .This study investigated the anti-apoptosis effect of alpha lipoic acid (α-LA) and its possible mechanism in the mouse model of traumatic brain injury. Methods Seventy-two healthy male ICR mice were randomly divided into four groups of 18 each:sham operation +vehicle, sham operation +α-LA, trauma +vehicle, and trauma +α-LA.The model of traumatic brain injury was made by weight-dropping.The animals in the α-LA groups were treated with intragastric α-LA at 30 minutes after surgery, while those in the vehicle groups with oral dimethyl sulfoxide in corn oil .At 48 hours after treatment , brain samples were collected from the mice for determining brain edema , measuring the expressions of cytochrome c , Bcl-2-associated X protein ( Bax ) , and caspase-3 in the mitochondria and cytosol of the brain tissue by Western blot and immunohistochemistry respectively , and detecting the survival of the neurons and apoptosis of neural cells in the cortical area by Nissl staining and TUNEL , re-spectively. Results The brain water volume , caspase-3 expres-sion, and neural cell apoptosis were markedly higher while the neuron survival remarkably lower in the trauma +vehicle group than in the sham operation +vehicle and sham operation +α-LA groups ( P<0.01).Compared with the mice in the trauma +vehicle group, those in the trauma +α-LA group showed significantly reduced proportion of water in the brain tissue ([79.89 ±0.55] vs [81.71 ± 0.66]%, P<0.05), expression of caspase-3 ([58.40 ±7.31] vs [47.42 ±7.74]%, P<0.05), and apoptosis of neural cells ([59.63 ±8.61] vs [44.86 ±7.32]%, P <0.05), but increased survival rate of neurons ([44.45 ±10.56] vs [57.46 ± 11.01]%, P<0.05).The expression of cytochrome c in the mitochondria was remarkably decreased and that of Bax markedly in -creased in the trauma +vehicle than in the sham operation +vehicle and sham operation +α-LA groups (P<0.01). Conclusion Alpha lipoic acid has a protective effect against traumatic brain injury in mice , probably by inhibiting the apoptosis of neural cells through the mitochondrial pathway .

Objective To examine expression of PTEN gene in ovarian cancer cisplatin-sensitive cell line OV2008 cells and cisplatin-resistant cell line C13K cells,and evaluate the effect of wild-type PTEN gene on reversing cisplatin-resistance of C13K cells and underlying mechanisms.Methods The expression of PTEN mRNA and protein in OV2008 and C13K cells were detected by semi-quantitative RT-PCR and western blot.Recombinant eukaryotic expression plasmid containing human wild-type PTEN gene was transfected into C13K cells by lipofectamine 2000.The expression of PTEN mRNA was monitored by RT- PCR and the expression of PTEN,protein kinase B(AKT),phospho-AKT(p-AKT)protein were analyzed by western blot in PTEN transfected and untransfected C13K cells.Proliferation and chemosensitivity of cells to cisplatin were measured by methyl thiazolyl tetrazolium(MTT),and cell apoptosis was detected by flow cytometry after treatment with cisplatin.Results(1)The expression of PTEN mRNA and protein(1.02 ?0.05,1.02?0.07)in OV2008 cells were significantly higher than those in C13K cells,which were 0.45 ?0.03 and 0.55?0.03 respectively(P

Objective To analyze the resistance genes in a muhidrug resistant Klebsiella pneumoniae (MDRKP) strain.Methods A MDRKP strain was isolated from bronchoalveolar lavage fluid in Pediatric Intensive Care Unit of Children's Hospital Affiliated to Soochow University in February 2012.Acquired resistance genes to beta-lactams,aminoglycosides,quinolones,ompK35 and ompK36 gene for outer membrane porin protein,and carbapenems targeting PBP2 gene were analyzed by PCR and verified by DNA sequencing.Results Acquired resistance genes TEM-1,SHV-1 to beta-lactam antimicrobial agents and aac(6′)-I b to aminoglycoside antimicrobial agents were positive in the strain of MDRKP.While 16S rRNA methylase,ompK35 and ompK36 genes for outer membrane porin protein were negative.Compared with susceptible strains,there were 9 synonymous mutations in PBP2 gene sequence of this MDRKP strain,but the amino acid sequences were the same.No mutation in quinolone resistance determining region (QRDR) was observed.Conclusion The multidrug resistance of the isolated Klebsiella pneumoniae strain may be related to 2 kinds of beta-lactam acquired resistance genes,1 kind of aminoglycoside acquired resistance gene,ompK35 and ompK36 genes defects and synonymous mutation in PBP2 gene.

AIM: To design,prepare and screen out functional small interfering RNA(siRNA) for specifically silencing proliferating cell nuclear antigen(PCNA) gene expression and effectively inhibiting cell proliferation on human hepatoma cell line SMMC-7721,human gastric carcinoma cell line SGC-7901 and human colorectal carcinoma cell line Caco2.METHODS: PCNA siRNA was designed based on previous studies about design guidelines and synthesized in vitro by T7 Mega short script reaction kit according to the manufacturer's instructions.After purification,the integrities of siRNA were identified through 19% denaturing polyacrylamide gel electrophoresis,and the concentrations of the generated siRNA were measured by testing the absorbance at 260 nm using a spectrophotometer.Four synthesized double-strand siRNA were transfected into three types of carcinoma cell lines by LipofectamineTM2000 reagent,respectively.WST-8 assay was employed to examine the proliferative inhibitions of these three cell lines.The subsequent alterations on PCNA mRNA and protein levels were determined by semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) and immunocytochemistry,respectively.RESULTS: 3 sequences,No.2,No.3 and No.4 PCNA siRNA showed effective inhibitions on tumor cells among the 4 candidate siRNA,and a single dose of 50 nmol/L of No.2,No.4 PCNA siRNA showed the most effective inhibitory rates as more than 62% at 48 h after transfection.50 nmol/L of No.2 and No.4 PCNA siRNA transfection caused 72% decrease of PCNA mRNA level and almost completely loss of the protein in human Caco2 cells.CONCLUSION: No.2 and No.4 PCNA siRNA have been screened out in this study,which show the capability to effectively down-regulated PCNA expression and significantly inhibit the proliferation of carcinoma cells.The optimal concentration is 50 nmol/L and satisfactory effects are achieved 48 h after transfection in vitro.

Objective To explore the changes in cardiac index and oxygenation index in sepsis piglets after nitric oxide (NO) gas inhalation. Methods A piglet model of sepsis was induced by intravenous infusion of Gram-negative bacterial endotoxin (LPS), then the piglets were randomly divided into two groups. NO group (n=8) was administered with inhaled nitric oxide of 80ppm via volume control (VC) mechanical ventilation for one hour, while the control group (n = 4) received mechanical ventilation with VC and was observed for one hour to assess the stability of the model. The parameters of oxygenation and hemodynamics were measured by PICCO and arterial blood gas analysis every fifteen minutes for one hour. Results Injection of endotoxin induced a stable pig model of sepsis. PH, HCO3-, arterial oxygen pressure (PaO2), mean arterial pressure (MAP) and cardiac index in this model were significantly lower the baseline values (P < 0.01). Arterial oxygen pressure and cardiac index were significantly higher in N0 group than in the control group (P<0.01). Heart rate (HR), central venous pressure (CVP), global end-diastolic volume index (GEDI) and intrathoracic blood volume index (ITBI) did not significantly differ between NO group and the control group. Conclusions Inhalation of nitric oxide gas can significantly improve oxygenation and cardiac function in piglets with sepsis.

Objective:To develop and optimize a novel assay for determination of cytotoxicity based Calcein -AM release.Methods:The target cells stained by Calcein-AM dye,then effectors and targets were incubated at E/T ratios from 30∶1-1∶1 for 4 h at 37℃,and the supernatant of reactions were detected by Fluorescence-Measurement to analyze specific cytotoxity.Results:The optimal excitation and emission wave lengths of Calcein were 485 nm and 515 nm.Dilutions of target cells stained by Calcein-AM had a linear relationship with measured fluorescence values.The Calcein-AM dye used to stain the living cells was shown to have a low spontaneous leakage rate-less than 15% in 4 hours at 37℃.Cytotoxicity activity of CIK showed a significant and positive correlation with E/T ratio when incubated at 4 h.Conclusion:The developed cytotoxicity test by Calcein-AM release is accurate and can avoid the application of radioactive reagents.

Objective To observe the primary clinical effect of concurrent radiochemotherapy for patients with nasal cavity natural killer (NK)/T cell lymphoma and to analyze the prognostic factors.Methods 31 primary untreated patients with stage Ⅱ nasal cavity NK/T cell lymphoma were enrolled for this study.All patients underwent concurrent radiochemotherapy with intensity-modulated radiotherapy technique + asparaginase based chemotherapeutic agents and adjuvant chemotherapy.Results The main toxicities were mouth mucocitis,myelosuppression and xerosmia at grade 1 or 2.31 patients achieved good clinical shortterm effect with high local complete remission rate at the 3rd month after radiotherapy [83.9 ％ (26/31)],and the 2-year overall survival rate was 77 ％.Univariate and multivariate analysis suggested IPI score and clinical short-term effect were the significant independent survival prognostic factors (P ＜ 0.05).Conclusions Concurrent radiochemotherapy for stage Ⅱ nasal cavity NK/T cell lymphoma can be well tolerated by patients with mild toxicities,and can improve both clinical short-term effect and overall survival by high local complete remission rate.IPI score and clinical short-term effect are the important survival prognostic factors.

Objective To study lymph node micrometastasis in N0 colorectal cancer patients and its clinical significance. Methods In this study, 548 lymph nodes obtained from 62 cases of No colorectal cancer undergoing curative operation were examined by fluorescent quantity polymerase chain reaction assay for the expression of cytokeratin 20 (CK20) mRNA. Results Micrometastasis was detected in 55 lymph nodes (10.0% ) of 24 cases (39%). According to lymph node anatomical locations, micrometastasis was identified in 15. 8%, 5.0% and 3.3% lymph nodes in group Ⅰ ,Ⅱ and Ⅲ (grouped by the distance from the tumor), respectively. Micrometastasis was correlated with invasion depth of primary tumor, but was notrelated to gender, age, tumor size, tumor site and differentiation. Conclusions The expression of CK20mRNA in lymph nodes in patients with No coloreetal cancer could be used to improve the accuracy of clinical staging, and provide information for rational adjuvant therapy.

Objective To study the changes of anorectal motility and rectal sensation in the elderly patients with ulcerative colitis (UC). Methods The anorectal motility and rectal sensation were investigated by Medtronic PC-Polygraf HR made by Sweden in 35 non-elderly patients versus 19 elderly patients with UC, and 20 non-elderly healthy subjects (HS) and 28 elderly HS were as control group. Results (1) The static pressure, pressure of anal sphincter and the maximal squeeze pressure of anal sphincter in non-elderly patients and elderly patients with UC showed no significant differences compared with those in non-elderly HS and elderly HS group (elderly patients with uc vs. ederly HA:t= 1.311,1.298,1.401;nonederly patients with uc vs. nonederly HS: t=1.294,1.299,1.322all P＞0.05). When abdominal pressure was increased, the net increased pressure of anal sphincter was (2.8±1.1) kPa in the elderly patients with UC, (2.9±1.3) kPa in the non-elderly patients with UC. The pressures were lower in two UC groups than in HS groups [elderly HS group:(3.8±1.2) kPa; non-elderly HS group:(3.9±1.2) kPa,elderly patients with uc vs. ederly HS:t=2.238,nonelderly patients with us vs. nonederly HS:t=2.243 all P＜0.05]. (2)The rectal lowest volume of sensory threshold, the maximal volume of tolerance and the maximal compliance were (85±30) ml, (180±69) ml, (26. 5±8.8) ml/kpa in elderly patients with UC and (65±15) ml, (170±58) ml, (22.6± 10. 3) ml/kPa in non-elderly patients with UC. They were lower than in each HS group [elderly HS group (95±31) ml, (205±78) ml, (32.9±12.9) ml/kPa; non-elderly HS group:(78±38) ml, (190±50) ml, (30.8± 15.2) ml/kpa, all P＜0. 01]. (3)The rectal lowest volume of sensory threshold, the maximal volume of tolerance and the maximal compliance in elderly patients with UC were higher than in non-elderly patients with UC (elderly patients with uc vs. elderly HS:t=3. 121,3. 135,3.146,nonederly patients with uc vs. non elderly HS: t= 3.162, 3.141, 3.188 elderly patients with uc vs. nonelderly patients with uc: t = 2. 246,2. 239,2. 240 all P＜ 0. 05). The rectal lowest volume of sensory threshold, the maximal volume of tolerance in elderly HS group were higher than in non-elderly HS group (ederly HS vs. t = 2. 328,2. 301 all P＜0. 05). Conclusions There are some anorectal motility disturbances in UC. Higher sensitivity, lower tolerance, lower compliance of rectum and weakened anal automatic control function in UC may be associated with diarrhea and frequent defecation. The rectal sensation threshold to volume stimulus is higher in elderly HS than in non-elderly HS group. The sensibility to volume ectasis of rectum is weakened and the survivability of rectum is increased in elderly patients with UC.