Chromatography ; methods ; DNA Fingerprinting ; Drugs, Chinese Herbal ; analysis ; Magnetic Resonance Spectroscopy ; Mass Spectrometry

Objective To observe the expressions of DNA methyltransferases (DNMTs) 1,3a and 3b in retinoblastoma (RB).Methods Sixty-two RB samples and six normal retinas were studied,including 17 poorly differentiated and 45 well differentiated samples; 16 invasive and 46 non-invasive samples.The expressions of DNMT1,3a,and 3b,and Ki-67 were detected using immunohistochemical analysis.Brown staining of nuclei was considered to represent the positive stain for DNMT1,3a and 3b,and ki-67,blue staining as negative.The level of high expression of nuclear staining was,positive cells in DNMT1≥65 ％,in DNMT3a≥60％ and in DNMT3b≥40％.The correlations of DNMT1,3a and 3b expression in RB samples,and MIB-1 labeling index were analyzed.Results Viewed under the light microscope,negative expressions of DNMT1,3a and 3b were demonstrated in normal retinas,however,positive expression was observed in RB samples,with 100％ in DNMT1,98％ in DNMT3a and 92％ in DNMT3b.Comparing well differentiated RB samples with poorly differentiated samples,significant differences were found in high expression of DNMT1 (x2 =12.57,P＜0.05) and DNMT3a (x2 =10.54,P＜0.05) ; also in the positive cells of DNMT1 (U=179,P＜0.05) and DNMT3a (U=198,P＜0.05).No significant difference was found comparing high expression (x2=1.5,P＞0.05) and positive cells (U=307,P＞0.05) of DNMT3b.When comparing invasive tumor tissues with non-invasive tumors,significant differences were shown between high expression (x2 =4.72,P＜0.05) and positive cells comparing DNMT1 (U=236,P＜0.05).No significant difference was shown in high expression (x2=3.53,0.84; P＞0.05) in DNMT3a and DNMT3b,or in comparison with positive cells (U=338,257; P＞0.05).The expression of DNMTs was positively correlated with the MIB-1 labeling index in RB tissues (R2=0.554,0.376,0.219; P＜0.05).Conclusion There are high expressions of DNMT1,3a,and 3b in RB.

Objective To observe the expression and relationship of high-mobility group A(HMGA) 1,HMGA2,MIB-1 labeling index (LI) and let-7 in retinoblastoma (RB).Methods Forty-four RB samples were studied,including 11 poorly differentiated samples,33 well-differentiated samples; eight invasive and 36 non-invasive samples.The expression of HMGA1,HMGA2 and MIB-1 LI in RB were analyzed by immunohistochemitry.The HMGA1,HMGA2 were scored on a scale of 0 to high expression.0: no expression ; low: 1％ - 10 ％ ; medium: 11 ％ - 50 ％ ; high: ＞50 ％.The MIB LI were scored on a scale of 0to high expression.O: no expression;low: 1％ - 40％;high: ＞ 40％.Semiquantitative reverse transcription-polymerase chain reaction was used to assay the let-7 expression level: ≥ 80％ showed no significantly decreased expression; 60％ - 79％ showed medium decrease in expression; ＜ 60％ highly decreased in expression.Results In 44 RB samples,there were 14 cases with no HMGA1 expression (32％),11 cases with low expression (25％),10 cases with medium expression (23％),and nine cases with high expression (20％).Expression level of HMGA1 was significantly higher in poorly differentiated RB than in well-differentiated RB (x2 =11.3,P＜0.01) ; however,no statistically significant difference was found between invasive tumors and noninvasive tumors (x2 -5.9,P＞0.05).There were 11 cases with no HMGA2 expression (25％),11 cases with low expression-(25％),nine cases with medium expression (20％),and 13 cases with high expression (30％).Expression level of HMGA2 was significantly higher in poorly differentiated and invasive RB than in well differentiated and noninvasive RB respectively (x2=20.9,8.7; P＜0.05).There were 4 cases with no MIB-1 LI expression (9％),18 cases with low expression (41％),and 22 cases with high expression (50％).Expression level of MIB1 LI was significantly higher in poorly differentiated RB than in well-differentiated RB (t=5.2,P＜0.05).Higher expression of MIB-1 LI was found in invasive tumors than in noninvasive tumors,with no significant difference (t=-1.1,P＞0.05).Twenty seven cases had no significantly decreased expression of let-7 (61％).There were eight cases with medium decreased expression (18％) and nine cases with highly decreased expression (21％).Correlation analyses revealed that MIB1 LI expression significantly correlated with HMGA1and HMGA2 proteins (r=0.327,0.602; P＜0.05).A significantly inverse correlation existed between let-7 expression and HMGA1,HMGA2 proteins and MIB-1LI respectively (r=-0.247,-0.310,-0.392; P＜0.05).Conclusions Overexpression of HMGA1,HMGA2 and MIB-1 LI and down regulation of let-7 were demonstrated in RB.Supplying let-7 to RB cells can possibly inhibit HMGA1 and HMGA2 expression.

Objective To analysis the clinical characteristics, treatment and prognosis for patients with cervical cancer with lymph lode of paraaortic and /or supraclavicular metastasis after combined treatment in two years. Methods Data of 20 patients with lymph lode of paraaortic and /or supraclavicular metastasis of cervical cancer were analyzed retrospectively. Results There was no evidence of lymph node of paraaortic and supraclavicular metastasis before the first combined treatments according to the MRI/CT/SPECT scans in these 20 patients. However, lymph lode of paraaortic and/or supraclavicular metastasis were seen in two years after treatments, the shortest is two months. One patient was progressing during treatment whose pathologic diagnosis is mucinous adenocarcinoma, died in 6 months from treatment start. There was only paraaortic metastasis in 15 patients, among these, 4 patients were found lymph lode of left supraclavical metastasis after retreatments in 3-6 months. Both paraaortic and left-supraclavical lymph node metastasis in 4 cases. Among these 20 cases, recurrence and to give up treatment, whose median survival time is 7.50 months, recurrence and to gain treatment, whose median survival time is 18.98 months, there was significant difference (x2=7.23, P<0.01). Conclusion Patients with cervical cancer with lymph lode of paraaortic and/or supraclavicular metastasis after combined treatment will obviously extend survival time after retretments, which has positive therapy significance.

[Objective]To study the best time for infusing antibiotics as prophylaxis against infection in hip arthroplasty.[Method]Ninty cases of hip arthroplasty were divided into 6 groups randomly with 15 cases each.Ampicillin(UNASYN,1.5g) and cefuroxime axetil(Zinacef, 2.0g) was administered by intravenous infusion respectively one day or one hour before operation or 30 minutes after the operation began.Blood samples from peripheral vein,cancellous bone of acetabulum when reaming,medullary canal of femur when rasping and catheter right after the operation were harvested.Serum Bactericidal Activities(SBAs) were determined by minim dilution technique and were compared between the groups.[Result]SBAs of UNASYN and Zinacef are similar to the bacteria most commonly causing infection after prosthesis implantation such as staphylococcus.Percentage at SBA≥1:8 of UNASYN and Zinacef is significantly more at the beginning of operation and when reaming acetabulum than the other groups(P

AIM: To qualitatively detect the concentration of recombinant human glucagon-like peptide-1(7-36)[rhGLP-1(7-36)] in human blood.METHODS: RhGLP-1(7-36) concentrations of standard and blood samples were detected by ELISA kits.RESULTS: The parameters of specificity,sensitivity,detection limits,reproducibiltiy and recovery can meet the requirements for pharmacokinetic studies.CONCLUSION: The concentration measuring method of rhGLP-1(7-36) in blood samples using ELISA kit produced by LINCO company can meet the detection methods of biological production,which can be used as a measuring method of rhGLP-1(7-36) concentration and for its pharmacokinetic studies.

AIM: To study the anti-cancer effects of octreotide on the proliferation of liver cancer cells. METHODS: The liver cancer cells SMMC-7721 were cultured in RPMI-1640 media with 10% fetal bovine serum, 100 U?ml -1 penicillin and streptomycin. 10 -5, 10 -4, 10 -3 and 10 -2 g?L -1 of octreotide were added and MTT colorimetric assay were used to detect the growth inhibition rate. DNA staining and cell cycle analysis were done at 0, 6, 12, 24 and 36 hours after medication when the concentration of octreotide was 10 -3 g?L -1. RESULTS: MTT colorimetric tests showed that octreotide suppressed the growth of liver cancer cells. 48 hours after medication, the cell growth inhibition rate was 9.33%, 12.70%, 19.70% and 20.93% when the octreotide concentration was 10 -5, 10 -4, 10 -3 and 10 -2 g?L -1 separately. Cell cycle analysis showed that the percentage of G 0-G 1 phase cells increased and the percentage of G 2-M phase cells decreased. CONCLUSION: Octreotide inhibits the cultured liver cancer cells proliferation in vitro, its mechanisms may be related to preventing the G 0-G 1 phase cells from going into G 2-M phase.

Objective To observe anti-sense phosphorothioate oligonucletide (ASPSODN) targeted directly to hTERT mRNA to its inhibiting effect on aimed gene and the influence on the telomerase activity, cellular proliferation, cell apoptosis of K562 cells. Methods Human leukemia cell line K562 was transfected with anti-sense oligonucleotide ASPSODN by liposome. The proliferation activity of K562 cell line was determined by using methyl thiazolyl tetrazolium assay, and telomerase activity was detected by TRAP-PCR-ELISA. Flow cytometry was adopted to examine apoptotic rate and cell cycle. RT-PCR was used to detect the expression of target gene hTERT mRNA. Results 0.6 μmol/L ASPSODN (0.42 ±0.16) was remarkably decreased the expression of hTERT mRNA, Telomerase relative activation was decreased by 52 %. According to 0.6 (μmol/L ASPSODN caused significant the inhibition of K562 cell growth. Apoptotic rate and cell cycle was examined by 0.6 μmol/L ASPSODN with flow cytometry. The cell apoptosis rate of 0.6μmol/L ASPSODN were 10.31 %. It showed the cells treated with 0.6 uU PSASODN arrested in G_1/C_0. The ratio of cells in G_2/M and S period was reduced. But there was no characteristic apoptosis peak. Conclusion ASPSODN targeted hTERT can inhibit the expression of target gene hTERT mRNA, and decrease the telomerase activity of K562 cells. ASPSODN can inhibit strongly the proliferation of K562 cell and induce cell apoptosis by decreasing telomerase activity.

Objective To ensure the quality of live attenuated measles vaccine on the market,viral titers were determined as the market surveillance test.Methods The measles vaccine was sampled from sites located in 24 provinces (cities,autonomous regions) of China,including provincial CDC,municipal CDC,county CDC,and primary users.The virus titer of samples was tested to evaluate the quality status of measles vaccine on the market.Results For all the 54 lots of measles vaccine sampled in this study,the virus titers were in compliance with the requirements of the China Pharmacopeia Vol.III (2005 edition).In comparison with the original test results provided by the manufacturers,the mean virus titer was decreased 0.2 lg?0.3.Conclusion Viral titers of measles vaccine batches sampled from the market were all in compliance with the requirements of the China Pharmacopeia Vol.III (2005 edition).It showed that the measles vaccine in China market has good stability,and the cold chain system for measles vaccine conveyance and storage could ensure the vaccine efficacy.

AIM: To determine the optimal operation method and reduce operative complications by retrospective analysis of outcomes of different operative manipulation for patients with congenital ectopia lentis ( CEL) .
●METHODS: A retrospective study ranged from Jan. 1, 2010 to Jan. 1, 2015 was conducted and 31 CEL patients (57 eyes) treated with different operations were taken into this study. The postoperative outcome, operation related complications was analyzed in terms of different operative manipulations.
● RESULTS: The most common types of CEL are idiopathic and Marfan - related CEL. All eyes ( 98%) benefited from operation except 1 Marchesani syndrome eye concurrent with optic atrophy induced by glaucoma. A total of 3 eyes ( 5%) were treated by phacoemulsification combined with intraocular lens implantation; 8 eyes ( 14%) were treated by phacoemulsification, capsular tension ring implantation combined with intraocular lens implantation, among these 8 eyes, 1 eye was treated with capsular tension ring implantation combined with suture fixation; 39 eyes ( 68%) were treated by lens excision and anterior vitrectomy through corneal incision combined with intraocular lens implantation with suture fixation, among these 39 eyes, 2 eyes were treated with trabeculectomy , but not combined with intraocular lens implantation; 5 eyes ( 9%) were treated by intracapsular cataract extraction and anterior vitrectomy combined with intraocular lens implantation with suture fixation; 2 eyes ( 4%) were treated by lens excision and vitrectomy through pars plana combined with silicon oil injection.
● CONCLUSION: Almost all patients can obtain satisfactory outcomes through various operative manipulations. Phacoemulsification combined with intraocular lens implantation was the first choice for patients with lens dislocation range less than 90°. Phacoemulsification, capsular tension ring implantation combined with intraocular lens implantation was recommended for 90° - 180°, and lens excision and anterior vitrectomy through corneal incision combined with intraocular lens implantation with suture fixation was more suitable for lens dislocation range more than 180°. The appropriate operative methods for different types of CEL patients is the key to success.