Background The filtering surgery is the main method of treating glaucoma.Fibrosis of filtering bleb is a key cause of failure of operation.The study about application of anti-scaring drug in filtering surgery is a hotspot.Objective Present study was to investigate the anti-scaring effect of paclitaxel aher trabeculectomy.Methods Thirty-two adult clean domestic rabbits underwent standardized trabeculectomy and randomly divided into 4 groups.Normal saline solution was used beneath the scleral flap during trabeculectomy for 3 minutes in 16 eyes of 8 rabbits as controls.0.3 g/L mitomycin C,0.2 g/L paclitaxel or 0.3 g/L paclitaxel was administered at the same way respectively in other 3 groups.Intraocular pressure (IOP) was measured,and eye numbers with function blebs were compared among 4 groups at the 4th,7th,14th and 28th day after surgery.The opening of filtration tunnel and the number of inflammatory cells were observed by hematoxylin and eosin staining,and proliferation of new collagen fibers was evaluated by Masson trichrome method.This study complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission(version 1988). Results No significant differences were found in the change of lOP among 4 groups before operation(F=0.54,P=0.83)and the 4th day aher operation(F=0.57,P=0.87).The IOP value was statistically lower in 0.3 g/L mitomycin C group,0.2 g/L paclitaxel group and 0.3 s/L paelitaxel group than the normal saline solution group(P＜0.05)with the lowest value in 0.3 s/L paclitaxel group in 7,14,28 days after operation(P＜0.05).Functional filtering bleb was seen in all the rabbit eyes in the 4th day after operation.In 7,14,28 days after operation,the number of eyes with functional bleb wag evidently more in 0.3 g/L mitomycin C group,0.2 g/L paclitaxel group and 0.3 g/L paclitaxel group compared with normal saline group(P＜0.05).The histological examination showed that the infiltration of inflammatory cells in filtering tunnel was much more obvious in normal saline solution group than the other groups with the most mild response in 0.3 g/L paclitaxel group.Masson trichrome revealed that proliferation of new collagen fibers in 0.3 g/L paclitaxel group was significantly decreased in comparison with those in other three groups at the 4th,7th,14th and 28th day after surgery(all P＜0.05). Conclusion Paclitaxel can inhibit the inflammatory response and collagen fibrosis and therefore open the filtering tunnel after it be used topically during the glaucoma trabeculectomy.

OBJECTIVETo establish a recombinant plasmid of CFP32 of Mycobacterium tuberculosis in E. coli, and to analyze its antigenicity.

METHODSRv0577 gene was amplified by polymerase chain reaction from genome of Mycobacterium tuberculosis, and then cloned into vector pMD18-T followed by the subclone into the expression vector pET21a. Recombinant CFP32 was expressed and purified. The antigenicity of the recombinant protein was analyzed by using Western-blot. The purified recombinant CFP32 protein was used as an antigen to screen the sera of 7 pulmonary TB patients (n = 97), as well as the other pulmonary disease patients (n = 25), and the clinically healthy controls (n = 38) by ELISA.

RESULTSRecombinant plasmid of CFP32 was established, and be expressed efficiently in E. coli BL21 (DE3). The relative molecular mass of the protein was about 300,000 by SDS-PAGE analysis. The protein purified by Ni-NTA was in a purity over 90%, which was confirmed by Western-blot analysis. ELISA analysis showed its sensitivity and specificity were 63.9% (62/97) and 96.8% (2/63) respectively.

CONCLUSIONThe recombinant expression plasmid pET21a CFP32 has been constructed and CFP32 proteins has been successfully expressed and be purified in E. coli and, ELISA analysis has identified the recombinant CFP32 as a candidate antigen for TB serodiagnosis.

Antigens, Bacterial ; blood ; Bacterial Proteins ; genetics ; immunology ; Cloning, Molecular ; Escherichia coli ; Gene Expression ; Humans ; Mycobacterium tuberculosis ; genetics ; isolation & purification ; Plasmids ; Recombinant Proteins ; Serologic Tests ; Tuberculosis, Pulmonary ; diagnosis ; microbiology

OBJECTIVETo investigate the effect of plant growth substances on induction and culture of callus from Rhodiola quadrifida and also to analyze salidroside contents in the callus.

METHODThe optimum combination of plant growth substances in MS solid medium for induction and culture of callus was established using orthogonal design. The contents of salidroside was analyzed by HPLC.

RESULTMS medium containing 2,4-D 1 mg x L(-1), NAA 2 mg x L(-1), 6-BA 0.5 mg x L(-1) and KT 0.1 mg x L(-1) could induce the callus from R. quadrifida most effectively;the induction rate was 83.3%. The optimized combination of plant growth substances for callus subculture was 2,4-D 1 mg x L(-1), 6-BA 0.1 mg x L(-1) and KT 0.5 mg x L(-1). The dry weight could reach 11.77 g x L(-1) when the callus was cultured in the optimum medium for 30 d and salidroside content was 0.28%.

CONCLUSIONThe quantities of plant growth substances required for induction and culture of callus are different in R. quadrifida. The callus could produce salidroside.

Culture Media ; Glucosides ; metabolism ; Phenols ; metabolism ; Plant Growth Regulators ; pharmacology ; Plant Stems ; growth & development ; metabolism ; Plants, Medicinal ; growth & development ; metabolism ; Rhodiola ; growth & development ; metabolism ; Tissue Culture Techniques ; methods

OBJECTIVETo detect the sequence variations frequently found within the N- and C-terminal regions of Epstein-Barr virus (EBV) LMP1 gene in nasopharyngeal carcinoma (NPC) and to study the underlying mechanisms.

METHODSFresh tumor tissues were sampled from 63 patients with untreated NPC encountered in Affiliated Tumor Hospital of Sun Yat-sen University, Guangzhou. The N-terminal region of EBV LMP1 gene was amplified with nested polymerase chain reaction (PCR), followed by XhoI enzyme digestion. Nested PCR was also employed to detect the 30 base pairs deletion within the C-terminal region. Four-colored fluorescence terminator sequencing method was applied for bi-directional solid-phase sequencing of the 8 representative PCR products in 4 cases of NPC. The DNA sequence within the N- and C-terminal regions of LMP1 gene was then analyzed.

RESULTSThere were 4 patterns of sequence variations, namely, wt-XhoI/wt-LMP1 (4 cases, 6.3%), wt-XhoI and XhoI-loss/del-LMP1 (4 cases, 6.3%), wt-XhoI/del-LMP1 (5 cases, 7.9%) and XhoI-loss/del-LMP1 (50 cases, 79.5%), detected in the 63 studied cases. Sequence analysis showed that the EBV LMP1 gene had underwent non-synonymous and synonymous substitutions, as compared with the prototype of B95-8 cells. The ratio of non-synonymous to synonymous substitutions was 2.25.

CONCLUSIONSXhoI-loss/del-LMP1 is the predominant sequence variation pattern of EBV LMP1 gene in NPC from Guangzhou. The XhoI-loss variation seems to develop on top of del-LMP1. When compared with the EBV LMP1 gene in peripheral blood B-lymphocytes of virus carriers and in preinvasive epithelial lesions (reported previously), it is likely that the sequence variation patterns of LMP1 gene may represent 4 different phases of intrahost evolution of EBV during nasopharyngeal carcinogenesis.

Adult ; Aged ; Base Sequence ; DNA, Viral ; genetics ; Deoxyribonucleases, Type II Site-Specific ; genetics ; Female ; Gene Deletion ; Genetic Variation ; Herpesvirus 4, Human ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Nasopharyngeal Neoplasms ; virology ; Point Mutation ; Sequence Analysis, DNA ; Viral Matrix Proteins ; genetics

AIMTo study the chemical constituents of the pine needles of Pinus massoniana lamb..

METHODSVarious chromatographic techniques were used to separate and purify. Their physico-chemical properties and spectral data (UV, IR, MS, 1H-1 H COSY, HMQC, DEPT, HMBC and ORD ect.) were measured for structure elucication.

RESULTSThree compounds were isolated from the n-BuOH fraction of water-extracts. Their structures were identified as massonianoside A (4), massonianoside A: (7S, 8R)-3, 4, 9'-trihydroxyl-3-methyoxyl-7, 8-dihydrobenzofunan-1'-propanolneoligan-9-O-alpha-L-rhamnopyranoside, massonianoside C (5), (7S, 8R)-9,9'-dihydroxyl-3,3'-dimethyoxyl-7,8-dihydrobenzofunan-1'- propanolneoligan-4-O-alpha-L-rhamnopyranoside and cedrusin-4-O-beta-glucoside (6), (7S, 8R)-3',9,9'-trihydroxyl-3-methoxyl-7,8-dihydrobenzofunan-1'- propanolneoligan-4-O-beta-D-glucopyranoside.

CONCLUSIONCompound 4 and 5 are new compounds.

Benzofurans ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Molecular Structure ; Pinus ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry

To optimize the belt drying process conditions optimization of Gardeniae Fructus extract from Reduning injection by Box-Behnken design-response surface methodology, on the basis of single factor experiment, a three-factor and three-level Box-Behnken experimental design was employed to optimize the drying technology of Gardeniae Fructus extract from Reduning injection. With drying temperature, drying time, feeding speed as independent variables and the content of geniposide as dependent variable, the experimental data were fitted to a second order polynomial equation, establishing the mathematical relationship between the content of geniposide and respective variables. With the experimental data analyzed by Design-Expert 8. 0. 6, the optimal drying parameter was as follows: the drying temperature was 98.5 degrees C , the drying time was 89 min, the feeding speed was 99.8 r x min(-1). Three verification experiments were taked under this technology and the measured average content of geniposide was 564. 108 mg x g(-1), which was close to the model prediction: 563. 307 mg x g(-1). According to the verification test, the Gardeniae Fructus belt drying process is steady and feasible. So single factor experiments combined with response surface method (RSM) could be used to optimize the drying technology of Reduning injection Gardenia extract.
Chemistry, Pharmaceutical ; instrumentation ; methods ; Desiccation ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Gardenia ; chemistry ; Research Design ; Vacuum

Objective To explore the application of modified CT imaging scale in the assessment of prognosis of comatose patients with severe traumatic brain injury. Methods Retrospective analysis was performed on the clinical data of 90 comatose patients with severe traumatic brain injury; according to the prognosis,they were divided into conscious group (n=47) and coma group (n=43).The modified CT imaging scale was used to analyze the relationship between these CT imaging data and prognosis,and the relationship between these CT imaging data and severity of illness. Results The scores of CT imaging scale in comatose patients were significantly higher than those in conscious patients (P＜0.05). Patients with 3-5 scores in GCS enjoyed higher scores of CT imaging scale as compared with those with 6-8 scores (P＜0.05).The scores of modified CT imaging scale and the GCS scores in these 90 patients showed negative correlation (Spearman's correlation coefficient:r=-0.79,P=0.000). Conclusion The modified CT imaging scale is helpful in assessing the prognosis and guiding the clinical treatment of comatose patients with severe traumatic brain injury.

Objective To observe the clinical curative effect of Annaohuoxue capsules, a prescription of promoting blood circulation and removing blood stasis, on patients with hypertensive cerebral hemorrhage and explore its safety. Methods Ninety-three patients with hypertensive cerebral hemorrhage,admitted to our hospital from June 2010 to November 201 i,were chosen and randomly divided into Arnaohuoxue capsule treatment group (group A,n=31),Tiandantongluo capsule treatment group (group B,n=32) and placebo control group (group C,n=30); After 7 d of onset,treatments were given,respectively,for 21 d.Hemorrheology,scores of neurological impairment,Glasgow coma scale (GCS) scores were recorded and compared in these groups before and after treatment. Results The plasma viscosity,and aggregation index of R BCs in patients from group A were significantly lower than those in patients from group B (P＜0.05); statistically significant difference on the clinical efficacy was noted between each 2 groups (P＜0.05):the outcome of group A was the best,that of group B was better,and that of group C was the worst.The GCS scores in patients from group A were significantly higher than those in patients from group B and group C (P＜0.05) Conclusion Annaohuoxue capsules can improve the hemorrheology indices and neurological impairment,and decrease the GCS scores in varying degrees,and the treatment efficiency of Annaohuoxue capsules is much better than placebo.

Spinal muscular atrophy (SMA) is a disorder characterized by degeneration of lower motor neurons and occasionally bulbar motor neurons leading to progressive limb and trunk paralysis as well as muscular atrophy. Three types of SMA are recognized depending on the age of onset, the maximum muscular activity achieved, and survivorship: SMA1, SMA2, and SMA3. The survival of motor neuron (SMN) gene has been identified as an SMA determining gene, whereas the neuronal apoptosis inhibitory protein (NAIP) gene is considered to be a modifying factor of the severity of SMA. The main objective of this study was to analyze the deletion of SMN1 and NAIP genes in southern Chinese children with SMA. Here, polymerase chain reaction (PCR) combined with restriction fragment length polymorphism (RFLP) was performed to detect the deletion of both exon 7 and exon 8 of SMN1 and exon 5 of NAIP in 62 southern Chinese children with strongly suspected clinical symptoms of SMA. All the 32 SMA1 patients and 76% (13/17) of SMA2 patients showed homozygous deletions for exon 7 and exon 8, and all the 13 SMA3 patients showed single deletion of SMN1 exon 7 along with 24% (4/17) of SMA2 patients. Eleven out of 32 (34%) SMA1 patients showed NAIP deletion, and none of SMA2 and SMA3 patients was found to have NAIP deletion. The findings of homozygous deletions of exon 7 and/or exon 8 of SMN1 gene confirmed the diagnosis of SMA, and suggested that the deletion of SMN1 exon 7 is a major cause of SMA in southern Chinese children, and that the NAIP gene may be a modifying factor for disease severity of SMA1. The molecular diagnosis system based on PCR-RFLP analysis can conveniently be applied in the clinical testing, genetic counseling, prenatal diagnosis and preimplantation genetic diagnosis of SMA.
Child ; Child, Preschool ; China ; epidemiology ; Female ; Gene Deletion ; Genetic Predisposition to Disease ; epidemiology ; genetics ; Humans ; Incidence ; Infant ; Male ; Neuronal Apoptosis-Inhibitory Protein ; genetics ; Polymorphism, Single Nucleotide ; genetics ; Spinal Muscular Atrophies of Childhood ; epidemiology ; genetics ; Survival of Motor Neuron 1 Protein ; genetics

OBJECTIVETo compare the Epstein-Barr virus (EBV) infection rates and the frequencies of wt-LMP1 and del-LMP1 EBV variants detected singly or dually among the four types of nasopharyngeal carcinoma (NPC) and to illustrate the possible role of del-LMP1 gene in nasopharyngeal carcinogenesis.

METHODSEBER in situ hybridization was performed in 117 NPCs, including 48 non-keratinizing carcinomas (NKCs), 25 keratinizing squamous cell carcinomas (KSCCs), 5 adenosquamous carcinomas (ASCs), 6 mucoepidermoid carcinomas (MECs) and 33 adenocarcinomas (ACs). Nested PCR for demonstration of EBV LMP1 gene was performed on the tissue samples collected from 99 EBER-positive carcinoma cases and the peripheral blood mononuclear cells (PBMCs) of 53 healthy adults (HAs).

RESULTSAs indicated by EBER in-situ hybridization, the EBV infection rates in both of 48 NKCs and 25 KSCCs were 100%; and the infection rates of 11 ASCs/MECs and 33 ACs were 9/11 and 51.5% (17/33), respectively. Worthy to note was that most of the NKC cells were EBER-positive while only a small number of EBER-positive neoplastic cells could be found in 17 ACs. The percentage of del-LMP1 EBV variant detected singly in NKCs (85.4%, 41/48) was not only significantly higher than that in PBMCs of 46 HAs (8.7%, 4/46) but also significantly higher than those detected in KSCCs (16.0%, 4/25). The dual infection rate of wt-LMP1 and del-LMP1 variants detected in KSCCs (56.0%, 14/25) was significantly higher than that of NKCs (12.5%, 6/48). The majority of the EBV detected in AC tissues (12/17) and HAs' PBMCs (34/46, 73.7%) were of dual wt-LMP1 and del-LMP1 variants.

CONCLUSIONSThe EBV infection rates are significantly different among 3 major histological categories, namely, NKC/KSCC, ASC/MEC and AC. Though NKCs and KSCCs are always consistently associated with EBV, the single del-LMP1 EBV variant detected in NKCs is predominant over that in KSCCs and most of the KSCCs contain dual wt-LMP1 and del-LMP1 EBV variants. The EBV of the del-LMP1 variant might play a crucial role in carcinogenesis of NKC.

Adult ; Epstein-Barr Virus Infections ; epidemiology ; Female ; Gene Deletion ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; etiology ; virology ; Viral Matrix Proteins ; genetics