Objective To study whether solasodine hydrochloride (SBHL) could enhance the effect of arsenic trioxide in inducing apoptosis and affecting telomerase activity in cervical cancer HeLa cells. Methods Using cell culture methods, cervical cancer HeLa cells were cultured in vitro. The optimal concentration of SBHL was determined by MTT method from 0, 10, 20, 40, 80, 160, to 320 μmol/L. HeLa cells were grown in improved RPMI1640 supplemented respectively with arsenic trioxide(5 μmol/L As2O3), As2O3(5 μmol/L)+ SBHL( 40 μmol/L) and none (control group). The growth morphology of HeLa cells was observed under phase contrast microscopy after culture for 24, 48, and 72 h. Apoptosis of HeLa cells was determined under transmission electronic microscopy. The method of MTT was used to study the cell survival percentage. The technique of flow cytometry was used to measure cell cycle and cell apoptosis percentage. The method of tartrate-resistant acid phosphatase-enzyme linked immunosorbent assay (TRAP-ELISA) was used to determine telomerase activity of HeLa cells. Results Under phase contrast microscopy, in control group HeLa cells were round, densely packed; in As2O3 group the numbers of the cells were less, cell spacing increased; in As2O3 + SBHL group the cells shrinked significantly, nuclear fragmented as a petal-like, gap became larger. Under transmission electronic microscopy, there were rich microvillus on the cell surface in control group, cell intervals clear, immature connections, and the intervals did not close. The structure of the mitochondria in the cytoplasm was integrated. Most of the chromatin in the nucleus were, euchromatin and characteristics of apoptosis with heterochromatin increased and the chromatin condensed into masses, on the boundary of nuclear membrane. The microvillud on the cell surface were ruptured and decreased in As2O3 + SBHL group. The chromatin condensed into masses. The formation of apoptotic bodies was observed. The difference was statistically significant between groups in cell survival percentage at 24, 48, 72h(x2 = 10.39 , 13.88 , 17.21,respectively, all P < 0.05). Cell survival percentage in SBHL + As2O3 group (52.80%) was significantly less than that of As2O3 group(77.51%, x2 = 9.29, P < 0.05) at 72 h. In cell cycles, the difference was statistically significant between groups in C1 phase and S phase(F = 7.46,22.14, all P < 0.05), respectively. Compared with , control group[ (41.57 ± 1.56)%, (50.45 ± 2.37)%], cell percentages in S phase in As2O3 + SBHL group[(20.06 ± 4.98)%] and As2O3 group[(27.10 ± 5.32)%] were decreased(P< 0.05 or < 0.01), while cell percentage in C1 phase was increased[(58.70 ± 5.18)%, (69.67 ± 4.17)%, P< 0.05 or < 0.01]. The difference was statistically significant between groups in apoptotic percentage of HeLa cells (F = 4.01, P < 0.05). Compared with control group[ (1.18 ± 1.40)%], apoptosis percentage was significantly increased in As2O3 + SBHL group and As2O3 group [(21.08± 1.22)%, (6.04±2.53)%, P< 0.05 or < 0.01], respectively, and As2O3 + SBHL group was higher than As2O3 group(P < 0.01). The difference was statistically significant between groups in telomerase activity (F = 21.28, P< 0.05). Telomerase activity was inhibited in As2O3 group(1.214 ± 0.621) and As2O3A + SBHL group(0.865 ± 0.284) compared to control group (2.107 ± 0.057, all P < 0.05), and telomerase activity in As2O3 + SBHL group was lower than that of As2O3 group (P < 0.05). Conclusions SBHL enhances the effect of As2O3 in inducing apoptosis in HeLa cells, which is related to its inhibiting telomerase activity in HeLa cells.

Objective To study the diagnostic and prognostic values by magnetic resonance imaging(MRI) and computed tomography(CT) for investigation of infantile spasms(IS).Methods Fourty-two patients with IS were retrospectively reviewed by CT scan and MRI T1W,T2W and inversion recovery (IR) and MRA techniques.Results Fourteen cases were found abnormal in CT,including encephalatrophy,hemorrhage,gross malformation and lesions with underlying calcification;MRI studies of 24/28 cases showed that MRI was the most appropriate imaging technique in diagnosis of the underlying substrate of patients with IS and other epilepsies,particularly in periventricular leuko malacia(PL),delayedmyelination(DM),hypxic-ischemic encephalopathy(HIE),kernicterus,tuberous sclerosis(TS),hippocampal sclerosis(HS),brainstematrophy,heterotopia,corpus callosum and vascular malformation,et al.MRI was also valuable for determining the prognoses of IS,but it should be combined with the clinical symptom and ages. Conclusions MRI and CT are highly important for the investigation and treatment of patients with IS; MRI is much more sensitive to exploration of neuropathology of infatile spasms,such as PL,DM,HIE,kernicteus,HS,heterotopias and focally cortical dysplasia.

Objective To explore the characteristics of etiology,clinical,electroencephalogram(EEG) and prognosis of infantile spasms(IS) with focal seizures(FS).Methods The significance of age onset,seizure patterns and atteration,etiology,video-EEG(VEEG) and evolution of FS correlating to epileptic spasms(ES),which occurred in 12 cases with IS by means of clinical observation,cranial CT or MRI,VEEG monitory and follow up were investigated.Twelve cases were divided into group A,B,C according to the stages of FS occurring prece-ding,coinciding and following ES.Results Ten cases with IS were identified from focal cortical dysplasia,tuberous scleroses complex,temporal lobe cyst or scleroses and hypoxic-ischemic encephalopathy and others.Clinical manifestation presented FS at certain times during the course of the disease,and other characteristics of frequent attacks,asymmetric spasms or tonic spasms,some atypical seizures in the eyes and the head,motionless staring and focal motor seizures and other.The correlation of ES to FS occurred during one ictal episode as follows:FS→ES(6 cases),ES→FS→ES(1 case),ES→FS(3 cases).FS appeared more frequently,atypical,predominantly involving ocular,facial,oral movement or generalized convulsion,migrating or alternating seizures,associated with epileptic discharges of posterior parietal-occipital and parietal-temporal-occipital origins in group A and early period of group B;whereas in the late period of group B and group C,FS occurred less frequently and more stable,presenting complex partial seizures or secondarily generalized seizures originated mainly from frontal and surrounding areas in the lateralized or bilateral hemispheres.Regarding the natural evolution and development of IS,2 cases with early FS developed into IS;6 cases with IS evolved into Lennox-Gastaut syndrome or symtomatic generalized epilepsy,4 cases got into FS,1 case maintained in a peculiarly epileptic state with FS as well as ES until the operation at 4.5 years old,and the other case was not identified clearly.Conclusions IS coincided with FS is a special kind of aged-related FS associated with secondarily generalized seizures and epileptic encephalopathy.The multiple etiology,seizure patterns,ictal-interictal EEG,clinical evolution and prognosis of the disorder,indicate a complicated interaction of the immature cortico-subcortical abnormalities in the critical developmental period and thereafter,can be identified as a peculiar form of clinical epileptic syndrome.

Some studies on nocardiosis in cultured fish were critically reviewed,including pathogenic bacterium,taxonomy,pathogenicity,histopathologically,diagnosis,controlling method and immunology. Some problem and advice on this disease were suggested.

Stenotrophomonas maltophilia is among eight strain of broad resistance to antibacterial. It has been pay attention to global students, and is very difficulty in clinic treatment. Infection of Stenotrophomonas maltophilia is more familiar in ICU. Its test rate is 13.45%(62/461) in the study .The resistance character: IPM﹑CZO﹑CRO﹑AMP and MNO is highest(resistive rate 100%). ATM、GEN 、 KAN 、MEM 、CTX and PIP is much high(≧86.5%), LVX﹑GAT and FEP is high (=50%).Conclusion: Stenotrophomonas maltophilia should be monitored in the hospital. Used of antibacterial must be rational. Severe disinfection and isolation should been put in practice to the infected patients. CSL is very active antibacterial (active rate 90.6%) against Stenotrophomonas maltophilia in the study, and ought be selected headmost in treated its infection.

Objective To analyze the expression level of HSP72 and HSP73,the subtypes of HSP70, in peripheral blood lymphocytes from lung carcinoma patients on both basic and heat injury conditions,and to explore the significance of HSP70 in the development of lung cancer.Methods Lung cancer patients were selected'as experimental group,and the health people with similar age,gender,vocational history and inhabi- tation to the experimental group were chosen as control group.The blood lymphocytes from both groups were isolated,cultured and treated with heat injury at either 37℃or 42℃.The expressions of HSP72 and HSP73 in the isolated lymphocytes were determined by Flow Cytometry.Results There were much higher expressions of HSP72 and HSP73 in control group(21.97?2.40 vs 12.77?0.66)than which in experimental group(HSP72 19.0?2.12 vs HSP73 11.74?0.68,P

OBJECTIVETo explore relationship between volume of bone cement injection and concurrent of fracture after thoracolumbar osteoporotic vertebral fracture treated by percutaneous kyphoplasty (PKP).

METHODSFrom January 2006 to December 2008,68 patients with thoracolumbar osteoporotic vertebral fracture treated by PKP were retrospectively analyzed. Among them, 30 patients with less than 3 ml bone cement injection (mean 2.5 ml, low group), including 11 males and 19 females, with an average age of (85.0 +/- 8.5) years (ranging for 60 to 91); 38 cases with over 4 ml bone cement injection (mean 4.5 ml, large group), including 15 males and 23 females,with an average age of (86.0 +/- 9.2) years (ranging for 60 to 93). Factors of concurrent vertebral fractures were observed during follow-up.

RESULTSAll patients were followed up from 3.4 to 5.1 years with an average of 3.8 years. Thirteen patients (43.3%) co-occurred fracture in low group,among which strengthened concurrent vertebral fracture occurred in 1 case,upper and lower section adjacent vertebral fracture in 8 cases,distal segment of vertebral fracture in 4 cases; while 18 patients (47.3%) co-occurred fracture in large group,among which strengthened concurrent vertebral fracture occurred in 2 cases, upper and lower section adjacent vertebral fracture in 10 cases,distal segment of vertebral fracture in 6 cases. No significant difference between two groups (P > 0.05).

CONCLUSIONBone cement injection is not main influence factors for treating concurrent of fracture after thoracolumbar osteoporotic vertebral fracture by PKP. Concurrent fracture mainly relates with progress of osteoporosis, the volume of injection volume may appropriately over the volume of balloon.

Aged ; Aged, 80 and over ; Bone Cements ; Female ; Follow-Up Studies ; Humans ; Injections ; Kyphoplasty ; adverse effects ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Osteoporotic Fractures ; etiology ; Postoperative Complications ; etiology ; Retrospective Studies ; Thoracic Vertebrae ; injuries ; surgery

Lanosterol synthase is encoded by the erg7 gene and catalyzes the cyclization of 2, 3-oxidosqualene, which is a rate-limiting step of the inherent mevalonate (MVA)/ergosterol metabolic pathway in Saccharomyces cerevisiae. The intermediate 2, 3-oxidosqualene is also the precursor of triterpenoids. Therefore, the cyclization of 2, 3-oxidosqualene is the key branch point of ergosterol and triterpenoids biosynthesis. Down-regulation of 2, 3-oxidosqualene metabolic flux to ergosterol in S. cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway reconstructed by the synthetic biology approach. To construct erg7 knockout cassette harboring the loxP-Marker-loxP element, long primers were designed, which were homologous to the sequences of both erg7 ORF and plasmid pUG66. The cassette was transformed into diploid wild strain INVSc1 by LiAc/SS Carrier DNA/PEG method and then erg7 gene haploid deficient mutant was obtained by homologous recombination. The results of semiquantitative PCR and real-time quantitative PCR revealed that erg7 expression level in erg7 gene haploid deficient mutant is one time lower than that in wild strain. The results of TLC and HPLC showed that the ergosterol content in deficient mutant decreased to 42% of that in wild strain.
Chromatography, High Pressure Liquid ; DNA Primers ; Down-Regulation ; Ergosterol ; metabolism ; Haploidy ; Intramolecular Transferases ; genetics ; Polymerase Chain Reaction ; Saccharomyces cerevisiae ; genetics ; Squalene ; analogs & derivatives ; metabolism

Traditional herbal medicines, Panax ginseng, Panax quinquefolium and Panax notoginseng, attract our attention for their extensive and powerful pharmacological activities. Ginsenosides are the main active constituents of these medicinal herbs. The related glycosyltransferases involved in ginsenoside biosynthesis are the key enzymes which catalyze the last important step. Modification of ginsenoside aglycones by glycosyltransferases produces the complexity and diversity of ginsenosides, which have more extensive pharmacological activity. At present, ginsenoside aglycones and compound K have been obtained by synthetic biology. As the last step of ginsenoside biosynthesis, glycosylation of ginsenoside aglycones has been studied intensively in recent years. This review summarizes the basic strategies and research advances in studies on glycosyltransferases involved in ginsenoside biosynthesis, which is expected to lay the theoretical foundation for the in-depth research of biosynthetic pathway of ginsenosides and their production by synthetic biology.
Biosynthetic Pathways ; Ginsenosides ; biosynthesis ; Glycosyltransferases ; metabolism ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Synthetic Biology

Pichia pastoris is one of the most important systems used in the field of molecular biology for the expression of recombinant proteins. The system has advantages of high expression, high stability, high secretion, easy high-density fermentation and low cost. Many factors affect the expression of recombinant protein, such as gene copy number, codon usage preference, type of promoter, molecular chaperones, glycosylation, signal peptide and fermentation process. In this review, research advances of the above aspects are summarized, which lay a foundation for improving the expression of recombinant proteins in P. pastoris.
Fermentation ; Gene Dosage ; Glycosylation ; Molecular Chaperones ; Pichia ; metabolism ; Promoter Regions, Genetic ; Protein Sorting Signals ; Recombinant Proteins ; biosynthesis