To evaluate in vitro release and transdermal behaviors of Huoxue Zhitong gel, modified Franz diffusion cell methods was applied to investigate in vitro transdermal absorption of Huoxue Zhitong gel and the content of paeonolan in receptor fluid composed of PEG400%-95% ethanol-water (l:3:6)were determined by HPLC. The results were processed and different equations were fitted. The release law were in accordance with Weibull equation and the fitting equation was In[-1/(1 - Q)] = -0.790 51nt - 1.7012 (r = 0.9809). In 8 hours, cumulative release of paeonol was 85. 18% and the release rate was 2.827 µg . cm-2 h-1. Transdermal actions were consistent with zero-level model fit and the fitting equation was Q(t) = 1.7579t + 0. 7213 (r = 0.9991). In 8 hours, cumulative transdermal rate and transmission rate of paeonol was 54. 85%, 1. 820 µg . cm-2 h-1. So the Huoxue Zhitong gel had a good release and transdermal properties.
Acetophenones ; administration & dosage ; pharmacokinetics ; Administration, Cutaneous ; Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Gels ; Mice ; Skin Absorption

Objective:To explore the predictive value of renal resistive index (RRI) joint with semiquantitative power Doppler ultrasound (PDU) score to acute kidney injury (AKI) in non-septic critically ill patients.Methods:This prospective observational study enrolled non-septic critically ill patients admitted to the Emergency Intensive Care Unit of Cangzhou Central Hospital from January 2018 to August 2019. In addition to general data, RRI and PDU scores were measured with medical ultrasonic instrument within 6 h after admission. Renal function was assessed on the 5th day in accordance with kidney disease: Improving Global Outcomes criteria. The patients who progressed to AKI stage 3 within 5 days after admission were classified into the AKI 3 group, and the rest were classified into the AKI 0-2 group. The difference of each index was compared between the two groups in non-septic critically ill patients and patients with acute heart failure (AHF). Normal distributed continuous variables were compared using independent sample t-tests, whereas Mann-Whitney U tests were used to examine the differences in variables without a normal distribution. Categorical data were compared with the Chi-square test. Receiver operator characteristic curves were plotted to examine the values of RRI, PDU score, RRI-RDU/10 (subtraction of RRI and 1/10 of PDU score), RRI/PDU (the ratio of RRI to PDU score), and RRI+PDU (the prediction probability of the combination of RRI and PDU score for AKI stage 3 obtained by logistic regression analysis) in predicting AKI 3. Delong's test was used to compare the area under the curve (AUC) between predictors. Results:A total of 110 non-septic critically ill patients (51 patients with no AKI, 21 with AKI stage 1, 11 with AKI stage 2, and 27 with AKI stage 3) were recruited. Among them, there were 63 patients with AHF (21 patients with no AKI, 15 with AKI stage 1, 7 with AKI stage 2, and 20 with AKI stage 3). Among the non-septic critically ill patients as well as its subgroup of AHF, compared with the AKI 0-2 group, acute physiology and chronic health evaluation-Ⅱ score, sequential organ failure assessment score, arterial lactate concentration, mechanical ventilation rate, proportion of vasoactive drugs, 28-day mortality, serum creatinine, RRI, RRI-RDU/10, RRI/PDU, RRI+PDU, and rate of continuous renal replacement therapy were higher in the AKI 3 group, and urine output and PDU score were lower ( all P<0.05). As for non-septic critically ill patients, RRI/PDU [AUC=0.915, 95% confidence interval ( CI): 0.846-0.959, P<0.01] and RRI+PDU (AUC=0.914, 95% CI: 0.845-0.959, P<0.01) performed best in predicting AKI 3, and the AUCs were higher than RRI (AUC=0.804, 95% CI: 0.718-0.874, P<0.01) and PDU score (AUC=0.868, 95% CI: 0.791-0.925, P<0.01). The optimal cutoff for RRI/PDU was > 0.355 (sensitivity 92.6%, specificity 81.9%, Youden index 0.745). The predictive value of RRI-RDU/10 for AKI 3 (AUC=0.899, 95% CI: 0.827-0.948, P<0.01) was also better than RRI and PDU scores, but slightly worse than RRI/PDU and RRI+PDU, with statistically difference only between RRI and RRI-RDU/10 ( P<0.05). As for patients with AHF, RRI/PDU (AUC=0.962, 95% CI: 0.880-0.994, P<0.01) and RRI+PDU (AUC=0.962, 95% CI: 0.880-0.994, P<0.01) also performed best in predicting AKI 3, and the AUCs were higher than RRI (AUC=0.845, 95% CI: 0.731-0.924, P<0.01) and PDU score (AUC=0.913, 95% CI: 0.814-0.969, P<0.01) with statistically differences (all P<0.05). The optimal cutoff for RRI/PDU was > 0.360 (sensitivity 95.0%, specificity 90.7%, Youden index 0.857). The predictive value of RRI-RDU/10 for AKI 3 (AUC=0.950, 95% CI: 0.864-0.989, P<0.01) was also better than RRI and PDU score, but slightly worse than RRI/PDU and RRI+PDU, with statistically difference only between RRI and RRI-RDU/10 ( P<0.05). Conclusions:The combination of RRI and PDU score could effectively predict AKI 3 in non-septic critically ill patients, especially in patients with AHF. The ratio of RRI to PDU score is recommended for clinical application because of its excellent predictive value for AKI and its practicability.

WRKY transcription factor is one of the Zinc finger proteins which contains a highly conserved WRKY domain and is a family of the plant-specific transcription factor. The plasmid pET28a-SmWRKY1 harboring Salvia miltiorrhiza WRKY1 (SmWRKY1) gene was successfully transformed and expressed in Escherichia coli BL21 (DE3). The conditions on protein expression of SmWRKY1 in E. coli, including induction duration, temperature, IPTG concentration and the E. coli concentration were optimized. The results showed that the highest protein expression of SmWRKY1 was obtained at 24 hours after the E. coli was cultured in the presence of 0.2 mol x L(-1) IPTG at 20 degrees C with A600 values of 1.0-1.5. This recombinant histidine-tagged protein was expressed at 2.454 g x L(-1) as inclusion body, which was first extracted using urea, and then purified by Ni2+ affinity chromatography and identified by SDS-PAGE. The expression of SmWRKY1 in E. coli was further confirmed by western blotting analysis.
Blotting, Western ; Cloning, Molecular ; DNA-Binding Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; chemistry ; genetics ; metabolism ; Molecular Weight ; Plant Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Recombinant Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Salvia miltiorrhiza ; genetics

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.
Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; metabolism ; Molecular Weight ; Open Reading Frames ; genetics ; Panax notoginseng ; chemistry ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment

Objective To study the application effect of day-parting appointment for elderly hypertensive contracted outpatients in community. Methods In May 2018 two groups (experiment and control) of 103 elderly hypertensive contracted outpatients, aged between 60 and 80 and looked after by the team of family doctors, who had been diagnosed with hypertension and with medication for at least one year were set up.The experimental group used self-made community hypertension visiting card for appointments, and the control group used the original way of treatment.Six months later, comparison was made in blood pressure control, the number of outpatients, the time consumed and the satisfaction between the two groups. Results It was found in comparison that the blood pressure standard-reaching rate of the experimental group was better than that of the control group (P < 0.05);outpatients′ visits and time consumed were less than those of the control group, the time used in the clinic room was longer than that in the control group; the differences in these aspects between the two groups were significant (P < 0.05);the satisfaction of the overall outpatient perception, attitude of family doctors, treatment technology, visits control, dosage, cost and safety of medication in the experimental group were better than in the control group; there were significant differences in these aspects between the two groups (P < 0.05). Conclusion Community day-parting appointment proves to be more convenient, more time-saving and safer for outpatients, greatly improving the medical quality and satisfaction for community outpatients.

BACKGROUND: Thoracolumbar fracture is commonly seen in spinal injuries, which causes loss of stability of the spine, as well as spinal cord and nerve compression, even deformity and paralysis. The diagnosis and treatment of thoracolumbar fracture remain controversial.OBJECTIVE: To summarize the mechanism of thoracolumbar fracture based on finite element method, its classification and transpedicular screw fixation.METHODS: The first author retrieved CNKI and PubMed databases for the relevant literature published between January 2000 and December 2016. The keywords were "finite element method, thoracolumbar spine fracture,transpedicular screw fixation" in Chinese and English, respectively.RESULTS AND CONCLUSION: (1) The finite element analysis method can simulate the mechanism of thoracolumbar fracture and provides a reference for the studies on the occurrence, development and treatment of thoracolumbar fracture. (2) The classification of thoracolumbar fracture is beneficial for planning a rational treatment strategy and evaluating prognosis. (3) Compared with the traditional screw fixation, the transpedicular screw fixation holds advantages in biomechanical stability and postoperative correction effect. (4) There are various classifications for thoracolumbar fracture; differences in severity and cartilage injury are difficult to simulate completely. (5) The finite element analysis method shows certain application limitations due to long learning curve and modeling time as well as complicated calculations.

OBJECTIVETo investigate the effect of X-rays on expression of caspase-3 and p53 protein in EL-4 cells and its implications in induction of apoptosis and polyploid cells.

METHODSMouse lymphoma cell line (EL-4 cells) was used. Fluorescent staining and flow cytometry analysis were employed for measurement of protein expression, apoptosis, cell cycle, and polyploid cells.

RESULTSThe expression of caspase-3 protein increased significantly at 8 h and 12 h, compared with that of sham-irradiated control (P<0.05, respectively) and the expression of p53 protein increased significantly at 2, 4, 8, 12, and 24 h, compared with that of sham-irradiated control (P<0.05-P<0.01) in EL-4 cells after 4.0 Gy X-irradiation. Apoptosis of EL-4 cells was increased significantly at 2, 4, 8, 12, 24, 48, and 72 h after 4.0 Gy exposure, compared with that of sham-irradiated control (P<0.05-P<0.001). G2 phase cells were increased significantly at 4, 8, 12, 24, 48, and 72 h (P<0.05-P<0.001). However, no marked change in the number of 8 C polyploid cells was found from 2 to 48 h after 4.0 Gy exposure.

CONCLUSIONThe expressions of caspase-3 and p53 protein in EL-4 cells are induced by X-rays, which might play an important role in the induction of apoptosis, and the molecular pathway for polyploid formation might be p53-independent.

Animals ; Caspase 3 ; metabolism ; radiation effects ; Caspases ; metabolism ; radiation effects ; Cell Line, Tumor ; Mice ; Tumor Suppressor Protein p53 ; metabolism ; radiation effects ; X-Rays

Objective To evaluate the value of combined optical coherence tomography (OCT) and intravascular ultrasound (IVUS) examinations in detecting coronary artery plaque during percutaneous transluminal coronary intervention (PCI).Methods CICT and IVUS examinations were performed on 30 diseased coronary vessels from 27 patients underwent PCI from Feb.2008 to Jul.2008.Results Seventeen vulnerable plaques (4 intima tearing which were not detected by IVUS),5 plaque rupture (1 out of 5 was detected by IVUS),5 thrombus lesions (1 out of 5 was found by IVUS),12 thin-cap lipid-rich lesions (2 detected by ivus) were detected by OCT in 22 lesions (without 8 lesions post DES stents).Analysis result of plaque burden by IVUS was superior to that obtained by OC T.In 8 DES stents (implanted for 6 months to 4 years).OCT detected 2 had severe restenosis,6 stents struts were completely covered with neointima without restenosis,1 stent had aneurysm-like dilatation.IVUS results were similar except for limitations on exactly detecting neointima post stenting.In 19 newly implanted stents.the incidence of stent underexpansion detected by OCT was 26.0% (same as that by IVUS).stent malappesition was 63.2% (10.5%by IVUS,P＜0.01),near stent tearing was 10.5% (not detected by IVUS),tissue prolapse between coronary stent struts was 52.6% (10.5% in IVUS,P＜0.05).Conclusions OCT imaging is superior to IVUS on detecting vulnerable plaques and change of structure around stents while IVUS is superior to OCT on estimating plaque burden in patients underwent PCI.

In order to construct RGD-mSAK mutant with reduced immunogenicity, and identify its biological activity after purification, mSAK gene fragment was amplified by over-lapping extension PCR. Then the gene was inserted into the prokaryotic expression vector pBV220 with P(R)P(L) promoters after confirmed by DNA sequencing; the expression plasmid pBV220-RGD-mSAK was constructed, and then was transformed into E. coli. DH5alpha. After temperature induction, the mutant Staphylokinase was over-expressed and much of protein was in the supernate of lysate, which is over 50% of total protein in the host. The protein was isolated and purified in Q-Sepharose FF, Sephacryl S-200 and SP, high purity protein was obtained and its purity was over 98%. The thrombolysis activity of the RGD-mSAK protein is 1.68 x 10(5) u/mg by fibrin plate assay, which is slightly higher than that of the wild-type, and antiserum titers raised against this protein in guinea pigs were much lower than those of wild-type SAK, determined by ELISA. In anti-platelets aggregation assay in vitro, the RGD-mSAK protein has obvious inhibition activity of platelet aggregation in low concentration comparing to the control group and wild-type SAK group. So the RGD-mSAK protein is a low immunogenicity, bi-function molecular with both thrombolysis activity and anti-embolism activity. It provided the basis for further research of RGD-SAK.
Animals ; Base Sequence ; Escherichia coli ; genetics ; metabolism ; Guinea Pigs ; Metalloendopeptidases ; biosynthesis ; metabolism ; Molecular Sequence Data ; Mutant Proteins ; biosynthesis ; genetics ; Oligopeptides ; metabolism ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; pharmacology ; Protein Engineering ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification

Objective To investigate the relationship between tumor necrosis factor α (TNF-α) rs1800629 polymorphisms and cervical cancer risk. Methods A case-control study was carried out including 552 patients with cervical cancer and 654 normal controls during the same period. TNF-α rs1800629 polymorphisms were examined by Taqman-Probe assay method. The association between the genotypes and cervical cancer was analyzed by Logistic regression models. Stata 11.0 was used for the Meta-analysis. Results Compared with the TNF-α rs1800629 GG genotype, individuals with GA, AA and GA/AA genotypes showed no significant changes in the risk of cervical cancer (all P>0.05). Further Meta-analysis on the relationship between the polymorphisms of TNF-α rs1800629 and cervical cancer also suggested that there was no significant correlation between the genetic variation and the occurrence of cervical cancer. Conclusion The polymorphisms of TNF-α rs1800629 may not be related to cervical cancer risk in Chinese population.