OBJECTIVE:To establish the quality standard of Liuwei dihuang pill (concentrated pill). METHODS:Radix Rehmanniae Preparata,Dioscorea opposita,Poria cocos and Alisma orientalis in Liuwei dihuang pill (concentrated pill) were identified by TLC and the contents of paeonolum and loganin were determined by HPLC.RESULTS:The TLC spots were clear and well-separated yet free of interference of negative sample.The linear ranges of paeonolum and laganin were 0.100 6～1.006 ?g (r=0.999 9) and 0.309 6～2.322 ?g(r=0.999 6),respectively,and their recovery rates were 98.2%(RSD=2.3%,n=6) and 95.3%(RSD=0.42%,n=6),respectively.CONCLUSION:The established standard is applicable for the quality control of Liuwei dihuang pill.

To evaluate the clinical characteristics of percutaneous coronary interventional (PCI) treatment in patients with chronic renal insufficiency, 116 patients with serum creatinine ≥141?mol/L and 352 patients who had normal serum creatinine level were included for the stady. It was found that the ineidences of using alcohol, acute myocardial infarction, hypertension, diabetes mellitus, as well as hospital stay days after PCI and the entire hospital stay days, and mean number of diseased vessels were significantly higher in renal insufficiency group compared with the control group. On the other hand, the value for HDL, the time for PCI operation, and the dose of contrast medium were lower in renal insufficiency group than in control group. The results suggested that patients with renal insufficiency were able to tolerate PCI if proper peri-operative care was observed, though they had more predisposing factors of arteriosclerosis.

AIM: To study the clinical curative effect of 2mm micro incision phacoemulsification combined with 23G minimally invasive vitrectomy for cataract and vitreoretinal diseases.METHODS: Retrospective analysis of 92 patients (99 eyes),including 49 male (53 eyes),43 female (46 eyes) with mean age was 57.1±1.9 years,in our hospital for cataract and vitreoretinal treatment of the disease from February 2013 to February 2016.All patients underwent 2mm micro incision phacoemulsification combined with 23G minimally invasive vitrectomy.Curative effect and complications were observed.RESULTS: Combined surgical procedures were carried out smoothly.posterior capsule rupture did not occurred.seven eyes were filled with BSS fluid,46 eyes with C3F8,49 eyes with intraocular lens at phase Ⅰ,21 eyes placed intraocular lens when silicone oil was removed.The visual acuity improved in 84 eyes (85%),unchaged in 15 eyes (15%).Postoperative complications included transient high intraocular pressure in 18 eyes (18%),anterior chamber reaction in 7 eyes (7%) and corneal edema in 8 eyes (8%).CONCLUSION: The 2mm micro incision phacoemulsification combined with 23G minimally invasive vitrectomy is a safe and effective surgical method with less injury,fewer complications.

Objective To study the effects of low-temperature environment on the contents of serum protein, serum calcium and blood sugar in rabbits. Methods The rabbits were divided into three experiment groups, which were exposed to environment at 2, -4 and -6 ℃ respectively and one control group exposed to environment at room temperature of 16 ℃. After exposure for 30～60 min, the peripheral blood samples were collected from rabbits for determination of levels of blood sugar, serum protein and calcium. The anal temperatures and ECG of rabbits were also examined. Results The levels of blood sugar, serum protein and calcium of rabbits in experiment groups decreased with the decrease of environmental temperatures, and revealed significant differences compared with those of control group(P

Objective To investigate the altered expression of TLR4 in alveolar macrophages of diabetic rats after lipopolysaccharide (LPS) stimulation and the effect of these changes on defending the infection. Methods Thirty-two male Wistar rats were divided into 4 groups, group A: the control group; group B: the diabetic group; group C: the LPS stimulated group; and group D: the diabetic group with LPS stimulation. TLR4 in alveolar macrophages were measured by immunocytochemistry, RT-PCR and Western blot analysis. Results The expressions of TLR4 in group B and group C were higher than that in group A (P < 0. 001). The expression of TLR4 in group D was obviously higher than that of group B and group C (P < 0.001). Conclusion The expression of TLR4 of diabetic rats was higher than that of normal rats and became more higher after LPS stimulation, which is indicated that diabetic bodies were in the proinflammatory state, the mechanism remains to be explored in detail.

Objective To evaluate T-lymphocyte subsets and sjTREC gene level in sepsis children and to provide a reasonable theoretical basis for immune regulation. Methods This prospective study was performed on children who were classified as sepsis group (n = 25), severe sepsis group (n = 23), and control group (n = 50). The T cell subsets were measured before the blood products,immune agents,and nutritional support were administrated. By real-time fluorescence quantitive-PCR method, the sjTREC levels of the patients with sepsis and healthy children were quantitatively detected respectively;then the sjTREC levels were absolutely quantified by two standard curve method. The results were demonstrated by sjTREC and endogenous reference gene (Cα) copies ratio (sjTREC/Cα× 2 × 1000) and statistically analyzed by SPSS 16. 0 software. Results The levels of CD3 +, CD4 +, and CD8 + T cells in severe sepsis group were lower than those in control group (P＜0.01). The level of sjTREC of severe sepsis group was 173.86 +38.37,which was significantly lower than those of sepsis group (345. 15 ± 66. 21) and control group (389. 42 ± 50. 20) (P ＜ 0. 01). Conclusion Children with severe sepsis have a range of T-lymphocyte subsets disorders and decreased thymic output function, so early immunotherapy can improve clinical outcome.

Blood Pressure ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Hypertension ; drug therapy ; Inflammation

AIM: To investigate the effect of fibrinogen (Fg), fibrin (Fb) and fibrin degradation products (FDPs) on the proliferation and migration of human vascular smooth muscle cells (VSMC).METHODS: The effects of Fg, Fb and FDPs on the proliferation of VSMC were observed by means of cell counting and MTT test, migration assays were performed using the wounding model and the transwell cell culture apparatus.RESULTS: Fg itself did not stimulate the proliferation of VSMC, but stimulated VSMC migration. Fb and FDPs both stimulated the proliferation and migration of VSMC, meanwhile the effect of Fb was in a dose-dependent manner.CONCLUSION: Fb, in particular FDPs, may play an important role by stimulating the proliferation and migration of VSMC in restenosis and atherogenesis.

In order to construct generally efficient cell immortalization vector, pTP-hTERT, we modified the traditional piggyBac (PB) transposon using artificial synthesis, PCR and enzyme digestion. The modified vector contained the necessary transposon elements, a PB transposase expression cassette, a co-expression selectable element and a human telomerase reverse transcriptase (hTERT) expression cassette. The co-expression selectable element had two markers, enhanced green fluorescent protein (EGFP) gene and puromycin-resistance (Puro) gene, linked by porcine teschovirus-1 2A peptide (P2A). To validate the functionality of vector elements, we transfected pTP-hTERT into HEK293 cell, selected the positive cell clones and then conducted RT-PCR, Western blotting (WB) and Tail-PCR, methylene blue staining and statistic analysis on selected cells. The results of sequencing and cell culture show that the pTP-hTERT was constructed successfully and the positive cell could be selected by puromycin. The WB results, P2A cutting EGFP and Puro fusion protein with high efficiency, reflected the selectable element worked. The sequencing result of Tail-PCR confirmed the vector integrated into the genome through transposition. The results of methylene blue staining and statistic analysis indicated the clone of positive cells triggered by pTP-hTERT significantly increased (P < 0.01) compared with control group. The construction of pTP-hTERT provides an efficient tool for establishing immortalized cell lines and a demonstration for building other eukaryotic plasmids.
Cell Line ; DNA Transposable Elements ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; HEK293 Cells ; Humans ; Plasmids ; Polymerase Chain Reaction ; Telomerase ; genetics ; Transfection

Objective To assess renal perfusion of chronic renal failure(CRF) at different stages in rabbits by contrast-enhanced ultrasound. Methods Fifteen rabbits were injected cationic bovine serum albumin(C-BSA) intravenously for 8 weeks to establish CRF models. Serum creatinine(Scr) and blood urea nitrogen(BUN) were determined before injection and 2,4,6,8 weeks after C-BSA injection respectively. The size of kidney was measured by two-dimensional ultrasound and contrast-enhanced ultrasound was performed on bilateral kidneys at the same time points. Renal perfusion was analyzed quantitatively with the time intensity curve. Results Renal cortical perfusion reduced started at 4weeks after injection,manifested as the peak signal intensity(PSI) of the time-intensity curve parameters decreased(P<0.05 or P <0.01). The speeds of perfusion and clearance of kidney were slower,showed as the time to peak intensity(PIT) and the time to half of peak intensity(HPT) delayed (P <0.05 or P <0.01). Compared with pre-injection, there were no differences in terms of the area under the curve(AUC) at 2,4 and 6 weeks after injection (all P> 0.05). And compared with pre-injection,2,4,6 weeks after injection, the AUC decreased at 8 weeks (P < 0.05 or P <0.01 ). The level of Scr and BUN of rabbits had increased since 6 weeks after injection (P< 0.05 or P <0.01). Two-dimensional ultrasound showed the renal volume was enlarged and the cortex was thickened from 2 weeks to 6 weeks after injection (P<0.05 or P <0.01). At 8 weeks after injection, the renal size had decreased as well (P < 0.05). These ultrasound changes were in accordance with its pathological changes. Conclusions Contrast-enhanced ultrasound in combination with time-intensity curve can quantitatively analyze the renal perfusion of CRF at different stages. The reduction of renal perfusion was earlier than the changes of routine laboratory indexes in rabbits with CRF. The haemodynamic changes of CRF rabbit were closely related with its pathological changes.