OBJECTIVETo investigate the clinical effects of double-plating internal fixation for the treatment of medial tibial plateau fractures with fracture-dislocation type through combined anterior and inverted "L" posteromedial knee approaches.

METHODSFrom February 2010 to September 2013, 17 patients with medial tibial plateau fractures with fracture-dislocation type were treated by double-plating internal fixation at our department. Anterior knee approach was adopted for medial mass reduction and fixation,while the inverted "L" posteromedial approach was used for posteromedial mass. There were 11 males and 6 females, with an average age of 42 years old (ranged, 21 to 65 years old). The efficacy were investigated by comparing pre- and post-operative radiographic data, and by evaluating clinical symptoms using the HSS score system.

RESULTSAll the patients were followed with a mean time of 18 months (ranged, 11 to 25 months). The tibial-plateau varus angle immediately after operation and at the latest follow-up were respectively (87.8 ± 4.2)° and (88.2 ± 4.6)°, and there was no statistical difference (P = 0.458). The posterior slope angle immediately after operation and at the latest follow-up were respectively (10.1 ± 3.7)° and (10.3 ± 4.1)°, and there was no statistical difference (P = 0.512). The knee joint HSS score was 86.7 ± 6.1. No patient had complications such as infection, skin necrosis, internal fixation loosening or loss of fracture reduction.

CONCLUSIONDoubl-plating internal fixation through combined approaches is an ideal operation method for medial tibial plateau fractures with fracture-dislocation type, because it can provide clear surgical field, satisfactory reduction, reliable fixation, and chance for early functional exercise.

Adult ; Aged ; Bone Plates ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Joint Dislocations ; surgery ; Male ; Middle Aged ; Tibial Fractures ; surgery

Objective To explore the dynamic change of serum leptin interleukin-6(IL-6),and interleukin-8(IL-8)during cerebral infarction,and to analyze the possible relationship between these factors and the severity of the disease.Method The levels of serum leptin,IL-6 and IL-8 were measured by radioimmunoassay in 60 patients with acute cerebral infarction,who were admitted to the hospital within 24 hours after stroke.Patients' neurologic impairment were evaluated by European Stroke Scale(ESS)at 1,3 and 7 days respectively after hospitalization.At the same time,another 30 normal adults were enrolled set as control group. Results The levels of serum Leptin,IL-8 in patients group after at 1 days,3 and 7 days were higher than those in the controls(P0.05).The ESS of these patients were 76.77?26.42,70.02?29.17,74.65?28.42 respectively,after 1,3 and 7 days.It shown that ESS simply correlated to the level of IL-8(P0.05).Statistic analysis was carried,but by using t test and linear correlation.(SPSS11.5 statistical package).Conclusion Leptin,IL-6 and IL-8 probably play a role in the inflammatory,reaction during acute cerebral infarction.The IL-8 may reflect the severity of the disease.

Objective To study the expression and change of neuropeptide substance P (SP) during the wound healing of deep partial thickness scalding in diabetic rats. Methods Eighty-four Wistar rats were randomly divided into diabetes mellitus group (n=42) and control group (n=42). Diabetic rat models were established by intraperitoneal injection of streptozotocin (STZ) in diabetes mellitus group, and those in control group were intraperitoneally injected with aseptic citrate buffer solution. Deep partial thickness scalding with diameter of 2 cm on the back were prepared in all the rats. The pre-scalding and post-scalding wound specimens of different time points were obtained, and the percentages of wound closure were calculated. The wound specimens were also obtained for immunohistological staining to compare the areas with positive staining of SP, and ELISA was employed to detect the expression of SP in the wound tissues. Results The percentage of wound closure was significantly lower in diabetes mellitus group than that in control group from 7 days post-scalding (P< 0.01). The areas with positive staining of SP in diabetes mellitus group were much smaller than those in control group at different time points, which was most significant on the seventh day post-scalding[(1 350.93±99.28) μm2 vs(1 715.86± 103.41) μm2](P < 0.01). The expression of SP in the wound tissues was significantly lower in diabetes mellitus group than that in control group at different time points, which was most significant on the seventh day post-scalding[(114.04±9.96) vs(143.39±8.94)](P<0.01). Conclusion The significantly lower expression of SP in wound site may be one of the causes of delayed wound healing in diabetic rats.

Acute Disease ; Adult ; Humans ; Infarction ; etiology ; Kidney ; blood supply ; Male ; Renal Artery Obstruction ; complications ; Thrombosis ; complications ; Tomography, X-Ray Computed

OBJECTIVETo investigate the effects of Salvianolic-acid B on p38MAPK signaling pathway and its transcriptional factor activated by Transforming growth factor b1 in rat hepatic stellate cells.

METHODSHepatic stellate cells were isolated from normal rat by in situ perfusion and Nycodenz density-gradient centrifugation method.TGFb1 (10 ng/ml), PD98059(50 mumol/L), SB203580(10 mumol/L) and SA-B (10-6 mol/L) were directly added to the medium of the isolated HSCs. Groups: (1)The detection of total p38, MKK3/6, MEF2A and MEF2C induced by TGFb1 in HSC: include control group, SA-B group, SA-B+TGFb1 group and TGFb1 group. (2)The detection of the phosphorylation of p38, MKK3/6 and a-SMA induced by TGFb1 in HSC: include control group, SA-B group, SA-B+TGFb1 group, TGFb1 group, PD98059 group, PD98059+SA-B group, PD98059+TGFb1 group and SA-B+PD98059+TGFb1 group. (3)The effects of SA-B on activity of MEF2 reporter and collagen a 1(I) reporter induced by TGFb1 in HSC: include mt group, wt group, TGFb1 group, SA-B+TGFb1 group, SA-B group, SB203580+TGFb1 group and SB203580 group. Total and phosphorylated p38 and MKK3/6, MEF2A, MEF2C and a-SMA were assayed by Western blot. HSCs were transfected with either MEF2 or collagen a1(I) luciferase reporter gene by Lipofectamine 2000 transfection method, Cellular extracts were assayed for both MEF2 and collagen a1(I) luciferase activities. Comparisons between groups were performed with Student-Newman-Keuls test.

RESULTSThe relative expression level of the phosphorylation of p38 of SA-B group is 0.33+/-0.05,obviously lower than control group(q=7.08, P less than 0.01); SA-B+TGFb1 group is 0.46+/-0.04, obviously lower than TGF b1 group(q=10.45, P less than 0.01); The relative expression level of the phosphorylation of MKK3/6 of SA-B group is 0.11+/-0.07, obviously lower than control group(q=3.944, P less than 0.05); SA-B+TGF b1 group is 0.28+/-0.07, obviously lower than TGFb1 group (q=7.91, P less than 0.01); The relative luciferase activity of MEF2 reporter of SA-B+TGFb1 group and SB203580+TGF b1 group is 2.93+/-0.09 and 2.50+/-0.05 respectively, both obviously lower than TGFb1 group(q=35.35 and 37.2, P less than 0.01); The relative expression level of MEF2C and MEF2A of SA-B group is 15.82+/-0.97 and 13.00+/-0.40 respectively, obviously lower than control group(q is 5.18 and 13.32, both P less than 0.01); SA-B+TGF b1 group is 13.40+/-0.72 and 20.47+/-0.83 respectively, obviously lower than TGFb1 group(q is 43.93 and 12.52,both P less than 0.01); The relative expression level of a-SMA of SA-B+TGFb1 group is 8.76+/-0.44, obviously lower than TGFb1 group(q=20.35, P less than 0.01); SA-B+SB203580+TGFb1 group is only 3.57+/-0.49, obviously lower than TGFb1 group(q=39.78, P less than 0.01); The relative luciferase activity of collagen a1(I) reporter of SA-B+TGF b1 group and SB203580+TGFb1 group is 1.61+/-0.05 and 1.42+/-0.07 respectively, obviously lower than TGFb1 group(q=26.4 and 27.62, both P less than 0.01).

CONCLUSIONSA-B could inhibit activation of HSC induced by TGFb1 through inhibiting p38MAPK signaling pathway in hepatic stellate cells.

Animals ; Benzofurans ; pharmacology ; Cells, Cultured ; Hepatic Stellate Cells ; drug effects ; metabolism ; MAP Kinase Signaling System ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism

To explore the antibacterial activity and mechanism of total alkaloids and berberine from Coptidis Rhizoma on Aeromonas hydrophila, and determine the effect of total alkaloids and berberine from Coptidis Rhizoma on minimum inhibitory concentrations, permeability and fluidity of cell membrane, conformation of membrane proteins and virulence factors of A. hydrophila. The results showed that both total alkaloids and berberine from Coptidis Rhizoma had antibacterial activities on A. hydrophila, with minimum inhibitory concentrations of 62.5 and 125 mg · L(-1), respectively. Total alkaloids and berberine from Coptidis Rhizoma could increase the fluidity of membrane, change the conformation of membrane porteins and increase the permeability of bacteria membrane by 24.52% and 19.66%, respectively. Besides, total alkaloids and berberine from Coptidis Rhizoma significantly decreased the hemolysis of exotoxin and the mRNA expressions of aerA and hlyA (P < 0.05, P < 0.01), the secretion of endotoxin and the mRNA expression of LpxC (P < 0.05, P < 0.01). The results suggested that the antibacterial activity of total alkaloids and berberine from Coptidis Rhizoma on A. hydrophila may be related to the bacteria membrane injury. They inhibited the bacterial growth by increasing membrane lipid fluidity and changing conformation of membrane proteins, and reduced the secretion of virulence factors of A. hydrophila to weaken the pathogenicity.
Aeromonas hydrophila ; drug effects ; genetics ; metabolism ; Alkaloids ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; metabolism ; Bacterial Toxins ; biosynthesis ; Berberine ; pharmacology ; Cell Membrane ; drug effects ; genetics ; metabolism ; Coptis ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Membrane Fluidity ; drug effects ; Rhizome ; chemistry

OBJECTIVETo explore whether rat mesenchymal stem cells (MSCs) can be induced to develop into hepatocytes and the role of the microenvironment of hepatocytes growth in inducing MSCs differentiating into hepatocytes in vitro.

METHODSMesenchymal stem cells were collected from the aspirates from femurs of SD rats by density gradient centrifugation and identified by flow cytometric analysis and alkaline phosphatase (ALP) staining. Rat hepatocytes were isolated by the modified two-step method described by Seglen. Two 6-well culture plates were piled up after the chambers' bottoms of the upper plate was removed. Then the upper and lower chambers were separated by a semi-permeable membrane. MSCs and hepatocytes of rats were plated separately in the upper and lower chambers of the two 6-well culture plates for co-culturing. MSCs cultured alone without co-culturing with hepatocytes served as controls. On days 1, 3, 7, 14, 21 and 28, mRNA of cytokeratin 18 (CK-18), alpha-fetoprotein (AFP) and albumin were detected by reverse transcriptase-polymerase chain reaction (RT-PCR), and immunocytochemistry staining of CK-18 AFP and albumin were also examined.

RESULTSThe shapes of MSCs co-cultured with hepatocytes changed and their sizes and numbers increased in the course of the culturing. When MSCs were co-cultured with hepatocytes for 2 weeks, colonies composed of polygonal cells resembling mature hepatocytes were found. In the controls, shapes of cells also changed and their sizes and numbers increased, but colonies composed of polygonal cells resembling mature hepatocytes were not found. Of the MSCs co-cultured with hepatocytes, on day 7, the mRNA of AFP was detected by RT-PCR, and it increased on day 14, and then decreased on day 21. mRNA of albumin and CK-18 were detected by RT-PCR from day 14 to day 28 in the co-cultured cells, but mRNA of AFP and CK-18 and albumin were not detected in the controls in the course of the culturing. Immunocytochemical analysis for CK-18, albumin, and AFP, showed positive staining reaction for AFP on day 7, for CK-18 and AFP on day 14 in the co-cultured cells but not in the controls.

CONCLUSIONSRat MSCs co-cultured with hepatocytes can differentiate into hepatocytes.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Coculture Techniques ; Hepatocytes ; cytology ; Male ; Mesenchymal Stromal Cells ; cytology ; Rats ; Rats, Sprague-Dawley

OBJECTIVEAfter performed symptom-limited maximum cardiopulmonary exercise testing (CPET) before and after acute alkalized blood, we repeated CPET with pure oxygen.

METHODSFive volunteers, 3hr after alkalizing blood room air CPET, re-performed CPET inhaling from Douglas bag connected with pure oxygen tank. We compared with those of room air CPETs before and after alkalized blood.

RESULTSAfter alkalized blood oxygen CPET had a similar response pattern as those of CPETs before and after blood alkalization. During the CPET, all breath frequency, minute ventilation and tidal volume at each stage were similar to those of CPETs before and after alkalized blood (P > 0.05),except there was a lower peak tidal volume than those of both CPETs and a slightly higher resting minute ventilation only than CPET after alkalized blood (P > 0.05). After alkalized blood, oxygen CPET, all PaO2 and SaO2 and most Hb were lower than those of both CPETs (P < 0.05). The pHa and [HCO3-]a were higher than those of CPET before alkalized blood (P < 0.05); but were not CPET after alkalized blood (P > 0.05). PaCO2 was similar to that of CPET before alkalized blood (P > 0.05), but was lower than that of CPET after alkalized blood at resting and warm-up (P < 0.05); then was similar to both CPETs at anaerobic threshold (P > 0.05); but was higher at peak exercise higher than those of both CPETs (P < 0.01). Oxygen increased 2,3 volunteers' workload and time at AT and peak exercises.

CONCLUSIONRespiratory response pattern to oxygen CPET after alkalized blood is similar to those of both CPETs before and after alkalized blood. The CPET response is dominantly depended upon metabolic rate, but not levels of pHa, PaCO2 and PaO2.

Blood Gas Analysis ; Exercise Test ; Humans ; Oxygen ; Respiratory Physiological Phenomena

Single nucleotide polymorphisms (SNP) is an important molecular marker in traditional Chinese medicine research, and it is widely used in TCM authentication. The present study created a new genotyping method by combining restriction endonuclease digesting with melting curve analysis, which is a stable, rapid and easy doing SNP genotyping method. The new method analyzed SNP genotyping of two chloroplast SNP which was located in or out of the endonuclease recognition site, the results showed that when attaching a 14 bp GC-clamp (cggcgggagggcgg) to 5' end of the primer and selecting suited endonuclease to digest the amplification products, the melting curve of Lonicera japonica and Atractylodes macrocephala were all of double peaks and the adulterants Shan-yin-hua and A. lancea were of single peaks. The results indicated that the method had good stability and reproducibility for identifying authentic medicines from its adulterants. It is a potential SNP genotyping method and named restriction endonuclease digest - melting curve analysis.
Atractylodes ; classification ; genetics ; DNA Restriction Enzymes ; metabolism ; DNA, Plant ; genetics ; Drug Contamination ; Genotype ; Lonicera ; classification ; genetics ; Plants, Medicinal ; classification ; genetics ; Polymorphism, Single Nucleotide

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