Objective To explore the differential diagnostic significance of clone analysis for multicentric occurrence (MO) and intrahepatic metastasis (IM) in hepatocellular carcinomas (HCCs).Methods Loss of heterozygosity (LOH) and microsatellite instability (MSI) were analyzed by microsatellite polymorphism test and the integration sites of HBV were assessed by Southern blot in each nodule of the HCCs. The clonalities were then compared between each nodule of the same patient and the diagnosis of MO or IM was concluded. Finally, the results based on clonality analysis were compared with those according to clinicopathological and imaging data. Results According to the results of LOH and MSI in 79 nodules and nontumorous tissue from 35 cases of mutiple HCCs, 5 (14.3%)and 29 cases (82.9 %) were divided into MO and IM, respectively. Both MO and IM presented simultaneously in 1 patient (2.9%). The integration sites of HBV could be analyzed in 77 nodules from 34 multiple HCCs. Among them, 6 (17. 6%) and 27 cases (79.4%) were divided into MO and IM, respectively. Both MO and IM presented simultaneously in 1 patient (2.9%). The classification results of microsatellite polymorphism test and HBV integration sites analysis demonstrated a significant positive correlation (rs = 0.909, P＜0.001). Nevertheless, neither the classification of microsatellite polymorphism test nor that of HBV integrate sites analysis was correlated with the discrimination according to clinicopathologic and imaging data (rs=0. 133, P=0. 468, rs =0. 262, P=0. 155, respectively). Recurrence in patients in the MO group occurred significantly later than that in IM cases who were diagnosed by clonality analyses (P=0. 001). Conclusion The clonality analysis based on the results of LOH and MSI or assessments of HBV integrate sites is useful for the differential diagnosis of MO and IM and their treatment and prognosis.

OBJECTIVE:To optimize the formulation of Dimemorfan phosphate tablets. METHODS:Using 60 min dissolution rate of dimemorfan phosphate as index,L9(34) orthogonal test was used to optimize the amount of starch,microcrystalline cellu-lose,croscarmellose sodium and concentration of HPMC E5 solution. The friability,hardness,60 min dissolution rate and main component were detected. The similarity of dissolution curves of Dimemorfan phosphate tablets was compared with that of imported tablets in 0.1 mol/L hydrochloric acid,pH 6.8 phosphate buffer,water and pH 4.0 acetate buffer. RESULTS:The optimized formu-lation of Dimemorfan phosphate tablet(1 000 tablets)was composed of dimemorfan phosphate 10 g,starch 60 g,microcrystalline cellulose 40 g,10% HPMC E5 solution and croscarmellose sodium 25 g. The friability,hardness,60 min dissolution rate and main component of 3 batches of Dimemorfan phosphate tablets prepared by optimized prescription were 0.42%-0.58%,9.8-10.5 kg,94.89%-96.21% and 99.21%-99.52%,respectively. In 4 dissolution mediums,similar factors f2 of dissolution curves between prepared tablets and imported tablets were above 50. CONCLUSIONS:Dimemorfan phosphate tablets were prepared successfully. The optimized formulation is rational. The dissolution behavior of prepared tablets is similar to that of imported tablets in vitro.

Objective To compare the clinical efficacy of single and double locking compressing plates(LCP)treatment for Schatzker type Ⅵ tibial plateau fractures.Methods Patients(n=90)with Schatzker type Ⅵ tibial plateau fractures were randomly divided into two groups according to the treatment methods (single LCP group and double LCP group;45 patients in each group) and analyzed retrospectively.Operative duration,intraoperative bleeding volume,postoperative wound infection,Rasmussen anatomical score and functional score were measured and compared between the two groups.Results More favorable results were achieved in the single LCP group than in the double LCP group in operative duration [(51.4 ± 4.03)h vs.(76.2±5.5)h(t=24.514,P＜0.001)]and intraoperative bleeding volume[(173.7±8.8) ml vs.(212.4 ± 4.9) ml (t =25.911,P ＜ 0.001,respectively)].Further more,three cases of postoperative woundin fection were seen in the double LCP group,while none was observed in the single LCP group.There was no significant difference in the percentage of satisfactory Rasmussenanatomical scores and functional scores between the two groups one year afteroperation(~ =-0.442,P=0.659).Conclusions Compared with double LCP fixation,single LCP fixation is more advantageous for the treatment of Schatzker type Ⅵ tibial plateau fractures.

Objective To investigate the effect of chymase on the proliferation of rat cardiac fibroblasts (CFs) and the role of transforming growth factor-?1 (TGF-?_1).Methods Cultured CFs of neonatal SD rats were isolated by trypsinization.Cell number and DNA synthesis were evaluated by MTT assay (A_(490) value) and [~3H]-deoxythy- midine [~3H]-TdR incorporation.The mRNA expression of TGF-?_1 in CFs was determined by RT-PCR.Results Chymase increased CFs numbers and [~3H]-TdR incorporation in a dose-dependent manner.The A_(490) value of CFs stimulated by 15,30 and 60 ng/mL chymase was 0.263?0.033,0.348?0.031 and 0.387?0.026,respectively, which were all significantly higher than that of control (0.201?0.019,P

In the present study, packaging system composed of pAAV-CMV-GFP, pAAV-RC and pHelper were transfected into human embryonic kidney 293 cells (HEK293 cells) mediated by polyethyleneimine (PEI) to explore an optimal transfection condition. Different total plasmid DNA dosages (1, 2, 3, 4, 5, 6 μg) and different PEI/Plasmid ratios (1:1, 3:1, 5:1, 7:1) were tested with detection of green fluorescence protein (GFP) with ImagePro Plus6. 0 Software. Then transfection efficiency of the optimized transfection system was further observed for different time periods(12, 24, 36, 48, 60, 72 h). The results showed that total plasmid dosage of 4 μg/well with PEI/plasmid ratio of 3 : 1-5 : 1 was an efficient transfection condition. Transfection efficiency-time curve was an S-shaped curve. Transfection efficiency reached a plateau at 60 h after transfection. The optimized conditions for PEI-mediated transfection at the optimal time result in enhanced transfection efficiency of triple plasmid into HEK293 cells.
Green Fluorescent Proteins ; HEK293 Cells ; Humans ; Plasmids ; Polyethyleneimine ; Transfection ; methods

Objective To compare the clinical results of percutaneous compression plating (PCCP)and Trigen short reconstruction intramedullary nail for intertrochanteric hip fractures. Methods During 2005 and 2008, the patients suffered with AO/OTA Al and A2 intertrochanteric hip fractures were divided into two groups; 36 fractures were treated with PCCP and 48 fractures with Trigen short reconstruction nail.During an average of (16.3±3.2) months follow-up, clinical evaluation involved visual analogue scale(VAS)score for pain in the 1st week, the 1st month, the 3rd month, the 6th month and the 12th month, and a Harris hip score one year post operation. Radiographs were examined for fracture healing-time, displacement scale of the neck screws and fracture impaction scale. All the complication in both groups was recorded. Results There were no difference in blood loss and operation time in both groups. The postoperative pain was significantly lower in the PCCP group in the initial three months after the surgery. Larger scale of fracture impaction and screw telescoping were seen in PCCP group. Also shorter healing time, higher Harris score results were achieved in PCCP group than those of Trigen short reconstruction nail group. Four peri-implant fractures occurred in Trigen short reconstruction nail group, which included one in the greater trochanter and three in the femoral shaft, but only one case need revision for bone displacement. In PCCP group, it was found that the superior neck screw was slightly displaced for tendency to cut-out in one patient. Conclusion Both PCCP and Trigen short reconstruction intramedullary nail can be successfully used to treat Al and A2 intertrochanteric hip fractures with minimal invasive technique. And the PCCP showed more rapid pain relief and bone healing, easily bony reduction and fewer complications.

OBJECTIVETo observe the effect of Chinese medicine and pharmacy (CMP) on the mortality of senile HIV/AIDS patients as adjunctive therapy.

METHODSHIV/AIDS patients of a certain rural area of Hanna Province, who were recruited in national CMP HIV treatment trial program (NTCMTP) in 2004, were enrolled as the CMP treatment group. HIV/AIDS patients in the same village without recruiting in NTCMTP were enrolled as the non-CMP treatment group. Data related to subjects were collected from the database of NTCMTP and National HAART Reporting System. Multiple regression analysis under Cox proportional hazard model was applied to examine the risk factors for death of senile HIV/AIDS patients.

RESULTSA total of 436 HIV/AIDS were enrolled in this study, 204 in the CMP treatment group and 232 in the non-CMP treatment group. There were 70 AIDS-relative deaths in the CMP treatment group, with 8-year mortality rate of 37.74%. There were 111 AIDS-relative deaths in the non-CMP treatment group, with 8-year mortality rate of 48.34%. The 8-year mortality rate was higher in the non-CMP treatment group than in the CMP treatment group (chi2 = 5.136, P < 0.05). Results of univariate Cox proportional hazards regression analysis showed the hazard ratio in the non-CMP treatment group was 1.41 times that of the CMP treatment group (P < 0.05). Result of multivariate Cox proportional hazards regression analysis showed the hazard ratio in the non-CMP treatment group was 1.44 times that of the CMP treatment group (P < 0.05). Besides, gender and marital conditions were significantly associated with death of HIV/AIDS patients.

CONCLUSIONCMP treatment was favorable to lower the mortality rate of senile HIV/AIDS patients, and its objective evaluation awaits for further prospective study.

Acquired Immunodeficiency Syndrome ; drug therapy ; mortality ; Alzheimer Disease ; therapy ; Antiretroviral Therapy, Highly Active ; Communicable Diseases ; Drugs, Chinese Herbal ; therapeutic use ; HIV Infections ; drug therapy ; mortality ; Humans ; Proportional Hazards Models ; Prospective Studies ; Risk Factors

To systematically evaluate the efficacy and safety of tripterygium glycosides (TG) combined with ACEI/ARB preparation in treating diabetic nephropathy stage IV. The computer retrievals were made in Cochrane Libarary, PubMed, Embase, SCI, Sinnomed, CNKI, Chinainfo and VIP, and hand retrievals were conducted for meeting and academic papers (updated to December 30, 2014), in order to collect randomized controlled trials and quasi-randomized control trials for TG combined with ACEI/ARB preparation in treating diabetic nephropathy stage IV and set the literature inclusion and elimination standards. Eligible literatures were included and evaluated according to standards in Cochrane Handbook. RevMan 5.3 and Stata 12.0 were used for a Meta-analysis. A total of 13 randomized controlled trials and quasi-randomized control trials involving 1119 patients with diabetic nephropathy were included. The Meta analysis result showed that compared with the control group, the combination group showed better effects in reducing the 24-hour urinary protein [MD = -0.84, 95% CI (-1.02, -0.66)], raising albumin [SMD = 0.98, 95% CI (0.81, 1.16)], the total efficiency [OR = 4.23, 95% CI (2.77, 6.46)] and the significant efficiency [OR = 5.35, 95% CI (2.70, 10.60)], with no statistical difference in Serum Creatinine between Both groups [MD = -0.82, 95% CI (-4.30, 2.66), P = 0.64]. However, the risk of adverse reactions increased by 7% [RD = 0.07, 95% CI (0.03, 0.12)]. The Egger's test showed no publication bias. Tripterygium Glycosides combined with ACEI/ARB in treating diabetic nephropathy stage IV is supper than the single administration of ACEI/ARB, with a good prospect in clinical application. Nevertheless, due to the small-size and low-quality samples in this study, more high-quality and large sample-size randomized controlled trials shall be conducted to verify the findings.
Angiotensin Receptor Antagonists ; administration & dosage ; Angiotensin-Converting Enzyme Inhibitors ; administration & dosage ; Diabetic Nephropathies ; drug therapy ; Drug Therapy, Combination ; Glycosides ; administration & dosage ; Humans ; Tripterygium ; chemistry

Objective To investigate the impact of polyclonal neural cell adhesion molecule antibody (P-NCAM-Ab) on the potency of botulinum toxin A (BTX-A).Methods Ninety male Sprague-Dawley rats were randomly divided into 3 equal groups:a normal control group,a BTX-A group and a P-NCAM-Ab group.The rats in the normal control group were injected with 100 μl of saline solution in their right gastrocnemius,while those in the BTX-A and P-NCAM-Ab groups were injected with 100 μl of BTX-A (0.5 U).In addition,the rats in the P-NCAM-Ab group were also injected with 100 μl of P-NCAM-Ab (the dosage was 20 U) at the same site on the 3rd day after the BTX-A injection.The rats' gastrocnemius muscle strength was evaluated with a self-made system for evaluating neuromuscular function before and after the toxin injection,on the 3rd day,as well as 1,2,4,6,8,10 and 12 weeks after the BTX-A injection.Any wet weight changes in the muscles were observed,and immunochemistry methods were employed to observe any structural changes in the motor endplates and nerve fibers at the different time points.Results After the saline injection,the average gastrocnemius muscle strength of the control group increased with time,while strength in the BTX-A and P-NCAM-Ab groups demonstrated a decrease in strength followed by a gradual increase.The average gastrocnemius muscle strength of the rats in the BTX-A and P-NCAM-Ab groups was significantly lower than that of the control group at all time points.Compared with the BTX-A group,the muscle strength of the P-NCAM-Ab group rats decreased further.Strength recovery in the BTX-A and P-NCAM-Ab groups was significantly slower than in the control group.The wet weight percentage in the BTX-A and P-NCAM-Ab groups at first decreased and then recovered with time.After the BTX-A injection,the average wet weight percentage of the P-NCAM-Ab group rats was significantly lower than that of the BTX-A group after 3 days,and 1,2 and 4 weeks.Karnovsky-Roots AchE staining showed that the motor endplates' color in the BTX-A and P-NCAM-Ab groups deepened gradually,though the color of the P-NCAM-Ab group was lighter than that of the BTX-A group at each time point.The mean optical density of the motor endplates' positive reaction area increased with time in both groups,but the P-NCAM-Ab group was lower than that of the BTX-A group at 1,2,4,8 and 12 weeks.Counting the nerve fibers dyed by gold chloride showed similar trends with both experimental groups significantly different from the control group.Conclusion P-NCAM-Ab can increase the potency of BTX-A and prolong its action.

Objective During zebrafish embryo gastrulation, the embryonic body plan is established by coordinated movements,including epiboly,involution,and convergence and extension(CE)movements. The aim of this study was to examine the expression patterns and the developmental functions of isthmin 1(ism1)during zebrafish gastrulation. Methods First, the expression of ism1 was examined using whole mount in situ hybridization during zebrafish embryonic development. Next,antisense morpholino oligonucleotide against ism1 and ism1 mRNA were injected into zebrafish embryos,respectively,to investigate its role in CE movements. Results ism1 is an early zygotic gene that is expressed in the dorsal side at 30% epiboly stage and in the blastoderm margin with the highest level in the dorsal organizer at shield stage as revealed by whole-mount in situ hybridization experiments. Compared with the wild-type embryos,either gain-or loss-of-function of ism1 leads to severe defects of CE movements. Conclusions Ism1 plays an essential role in CE movements during zebrafish embryo gastrulation.