Chitinases genes from Metarhizium anisopliae which is an important entomopathogenic fungus were considered one of the key factors to invade their hosts. One Metarhizium anisopliae HN1 strain was isolated and screened. A chitinase gene was amplified by RT-PCR from Metarhizium anisopliae HN1, The whole length of this gene was 1275bp,and the nucleotide sequence of the gene was 96% similarity to that of the M. anisopliae E6 accessed in GenBank ( AF02749). The gene has been registered in GenBank and its accession number is DQ011865. The gene was subcloned into prokaryon expression vector pET-22b( + ), transformed this recombinant expression plasmid into E. coli strain BL 21 and effective expressed. The SDS-PAGE analysis indicated that the recombinant protein was 42kDa which is same to the reported article. The expression level of recombinant protein was about 63. 3% of whole expressed proteins , And when recombinant E. coli were crushed by freeze and supersonic wave , the activity assay indicates that the chitinase expressed in bacteria possesses biological activity.

OBJECTIVETo observe the influence of Gandou Tablet I (GD I) on neurologic electrophysiology in patients of hepatolenticular degeneration (HLD).

METHODSVigilance-controlled electroencephalogram (VEEG), electromyogram (EMG) and brain-stem auditory evoked potential (BAEP) in 32 cases of HLD were examined before GD I treatment, and reexamined after 4 weeks of treatment in those with abnormal neurologic electrophysiology.

RESULTSAt the same time of clinical symptom improvement after treatment, the abnormal VEEG, EMG and BAEP also improved with the effective rate of 71.43%, 70% and 66.67% respectively, their constitutional ratio were different insignificantly, P > 0.05. The peak latency (PL) of III, V waves and interpeak latency (IPL) of I-III, III-V and I-V waves in patients before treatment were all greatly prolonged, compared with those in the healthy control group (P < 0.05 or 0.01), the IPL of III-V and I-V waves were markedly improved after treatment.

CONCLUSIONNeurologic electrophysiologic examination was valuable for complementary diagnosis and therapeutic effect evaluation of HLD. GD I could improve the abnormal neurologic electrophysiologic parameters.

Adolescent ; Adult ; Child ; Drugs, Chinese Herbal ; therapeutic use ; Electroencephalography ; drug effects ; Electromyography ; drug effects ; Electrophysiology ; Evoked Potentials, Auditory, Brain Stem ; drug effects ; Hepatolenticular Degeneration ; drug therapy ; physiopathology ; Humans ; Male ; Phytotherapy ; Tablets

Objective:To compare the effectiveness of fire needle versus Western medicine in the treatment of herpes zoster. Methods:Randomized controlled trials comparing fire needle with Western medicine in the treatment of herpes zoster were identified using 8 databases. A meta-analysis was performed using RevMan 5.3 software. Results:Eight trials involving 569 patients were included in this meta-analysis, and the results showed that fire needle was superior to Western medicine comparing the effective rate [risk ratio (RR)=1.13, 95％ confidence interval (CI): 1.06 to 1.20;P=0.0002], the visual analog scale (VAS) score [mean difference(MD)=–7.95, 95％ CI: –10.71 to –5.20;P<0.00001], time of pain disappearance (MD=–7.61, 95％CI: –9.38 to –5.84;P<0.00001), time of blister-stop (MD=–1.34, 95％CI: –1.51 to –1.18;P<0.00001), time of crusted scab (MD=–2.92, 95％CI: –3.62 to –2.23;P<0.00001), and time of scab off (MD=–4.64, 95％CI: –5.83 to –3.46;P<0.00001). In addition, a significantly lower incidence of postherpetic neuralgia was found in the fire needle group in 30 d (RR=0.23, 95％CI: 0.11 to 0.51;P=0.0002) and 60 d (RR=0.33, 95％CI: 0.12 to 0.91; P=0.03) after treatment. Conclusion:Fire needle has a favorable effect in increasing the effective rate, relieving pain, recovering skin lesions and decreasing incidence of postherpetic neuralgia in the treatment of herpes zoster. However, considering the limitations in this study, the findings should be interpreted cautiously.

Most chemical medicines have polymorphs. The difference of medicine polymorphs in physicochemical properties directly affects the stability, efficacy, and safety of solid medicine products. Polymorphs is incomparably important to pharmaceutical chemistry, manufacturing, and control. Meantime polymorphs is a key factor for the quality of high-end drug and formulations. Polymorph prediction technology can effectively guide screening of trial experiments, and reduce the risk of missing stable crystal form in the traditional experiment. Polymorph prediction technology was firstly based on theoretical calculations such as quantum mechanics and computational chemistry, and then was developed by the key technology of machine learning using the artificial intelligence. Nowadays, the popular trend is to combine the advantages of theoretical calculation and machine learning to jointly predict crystal structure. Recently, predicting medicine polymorphs has still been a challenging problem. It is expected to learn from and integrate existing technologies to predict medicine polymorphs more accurately and efficiently.

OBJECTIVETo establish methods for the determination of phillyrin and forsythoside A in Lianqiao Bingdu Qing capsule by RP-HPLC.

METHODThe determination of phillyrin was carried out on YWG-C18 column (4.6 mm x 250 mm, 10 microm), using acetonitrile-water (25:75) as mobile phase at a flow rate of 0.8 mL x min(-1) and detected at the wavelength 277 nm. The determination of forsythoside A was carried out with YWG-C18 column(4.6 mm x 250 mm,10 microm), using acetonitrile-water-Acetic acid (17:83:0.4) as mobile phase at a flow rate of 1.0 mL x min(-1) and detected at the wavelength 280 nm.

RESULTThe average recovery of phillyrin was 99.6%, RSD = 1.9% (n = 5). The average recovery of forsythoside A was 101.3%, RSD = 2.5% ( n = 5).

CONCLUSIONThe methods were simple and accurate and could be used to control the quality of the Lianqiao Bingdu Qing capsule.

Capsules ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Forsythia ; chemistry ; Fruit ; chemistry ; Glucosides ; analysis ; Glycosides ; analysis ; Plants, Medicinal ; chemistry

Objective To investigate the mutations ACTN4 and SYNPO genes promoter in sporadic primary focal segmental glomerulosclerosis (FSGS) and to analyze the role of mutations in FSGS. Methods The study consisted of 82 Chinese primary FSGS, including 39 females and 43 males, ranged from 12 to 76 years old. Seventy volunteers were selected as healthy control group. Genomie DNA was extracted from peripheral blood cells of FSGS patients and hair of patients' parents by polymerase chain reaction (PCR) and direct sequencing to analyze ACTN4 and SYNPO gene promoter mutations. Mutations were matched with GenBank and TRANSFAC software database (www.ncbi.nlm.nih.gov; www.genometix.de; www.gene-regulation, corn). Dual luciferase assay system was used to analyze the promoter region mutations, based on PGL3-Basie vector, pRL-SV40 and PCI2 cell line. Hair DNA of novel mutation patients' parents was sequenced. Expression of alpha-actinin-4 and synaptopodin in patients' kidney tissue was examined by immunofluorescence. Results Three patients with 1-34C>T, 1-590delA and (1-1044delT)+ (I-797T >C) +(1-769A >G) heterozygous mutations were found in ACTN4 gene promoter respectively, and two patients with 1-24G>A and 1-851C>T heterozygous mutations in SYNPO gene promoter respectively. The same mutations were not found in the control group of 70 healthy people. Except one patient accepting her parents' 1-1044delT and 1-797T>C mutated chromosome respectively, no same mutations were found in patients' parents. Protein expression of alpha-actinin-4 and synaptopodin was reduced in mutated patients' kidneys. Except 1-1044delT group, luciferase activity in mutated groups decreased. (1-1044delT)+(1-797T>C)+(1-769A>G) mutation was associated with poor outcome and patient with these mutations progressed to end-stage renal failure. Conclusion Mutations of ACTN4 and SYNPO gene promoters affect gene transcription and protein translation, which may contribute to the onset of sporadic primary FSGS.

AIMTo investigate the effects of praeruptorin C (Pra-C) on smooth muscle cell (SMC) hypertrophy, intracellular calcium ([Ca2+]i), nitric oxide (NO) content and influence on cellular signal transduction in isolated cultured rat smooth muscle cell (SMC).

METHODSHypertrophied smooth muscle cells (HSMCs) were induced by angiotensin II (Ang II), cell area was measured under inverted microscope. Nitric oxide (NO) concentration was measured using Griess method. [Ca2+]i was measured using Fura-2/AM. The responses to [Ca2+]i elevation stimulated by KCl (60 mmol.L-1 or norepinephrine (10 mumol.L-1) were observed by incubation with phorbol 12-myristate 13-acetate (PMA), staurosporine (ST), the agonist and inhibitor of protein kinase C (PKC), and pertussis toxin (PTX), the sensitive toxin of Gi.

RESULTSThe cell area of SMCs were decreased by 39.01% (P < 0.001) and NO content of SMCs were significantly increased in Pra-C + Ang II group. In presence of 60 mmol.L-1 KCl or 10 mumol.L-1 NE, [Ca2+]i of SMCs in Pra-C + Ang II group was significantly decreased than that of Ang II group (P < 0.001) and closed to the normal group. Incubation of SMCs with PMA, ST and PTX, [Ca2+]i of SMCs in Ang II group was increased by PMA and decreased by ST and PTX, but that of Pra-C + Ang II group was similar to the normal group.

CONCLUSIONThese findings suggest that Pra-C can reduce vascular hypertrophy in isolated rat HSMCs, and this is associated with improvement of SMCs [Ca2+]i level, NO content and cellular signal transdution of PKC and Gi.

Angiotensin II ; Animals ; Aorta ; pathology ; Calcium ; metabolism ; Calcium Channel Blockers ; pharmacology ; Cells, Cultured ; Coumarins ; pharmacology ; Female ; Hypertrophy ; chemically induced ; pathology ; Male ; Muscle, Smooth, Vascular ; drug effects ; Myocytes, Smooth Muscle ; drug effects ; pathology ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects

AIMTo synthesize eudistomin U and its 6-OCH3/Br derivatives and 5'-Br derivatives as antitumor agents.

METHODSUsing tryptamine and indole-3-aldehyde as starting materials, through condensation, Pictet-Spengler cyclization and dehydrogenation three steps, the alkaloids and its derivatives were prepared.

RESULTSThe structures of the compounds were determined by 1HNMR, MS and HRMS. Antitumor activity in vitro was tested.

CONCLUSIONEudistomin U and its derivatives were synthesized. The results showed that they all showed antitumor activities against mouse P388 strain.

Alkaloids ; chemical synthesis ; chemistry ; pharmacology ; Animals ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Carbolines ; chemical synthesis ; chemistry ; pharmacology ; Cell Division ; drug effects ; Cell Line, Tumor ; Indoles ; Leukemia P388 ; pathology ; Mice ; Molecular Structure ; Tryptamines

3 cm)and small lesions(diameter≤3 cm)were 80.6%(79/98)and 67.2% (45/67),respectively(P

Objective To establish serological detection methods of human herpesvirus 8 (HHV-8).Methnds Three potent antigenic fusion proteins.K8.1,ORF65 and ORF73 C of HHV- 8 were synthesized using E.coli system.The sera were detected using lhese antigenic proteins.The positive sera were from 12 patients with Kaposi's sarcoma and 32 patients with acquired immunodeficiency syndrome-related Kaposi's sarcoma.The negative sera were from 20 patients with cutaneous tumors and children under 15 years old.Western blot and enzyme-linked immunosorbent assay (EI.ISA)were employed to determine the immunogenicity of each recombinant protein and the sensitivity and specificity of ELISA using the complex antigens.Results Three types highly purified HHV 8 specific recombinant pro teins with potent antigenicity were successfully synthesized.The sensitivity of ELISA using the above complex antigens was significantly higher than traditional immuno-flurescent assay (IFA)detecting the positive and negative sera,whieh were 81.8%,34.4%,respectively.And the specificity of ELISA was 97.9%.Conclusion K8.1,ORF65 and ORE73 C are good candidate antigens for establishing HHV-8 serological detection methods,which have better sensitivity and specificity.