The medium of photo-synthetic bacteria synthesizing carotenoid was optimized with response surface methodology. The regression equation expressing the relationship between carotenoid yield and main components of culture medium was established by stepwise analysis. The R-squared in the model of regression equation was 0. 958, which meant the established equation could predict the carotenoid yield well at the range of factors in this design. It was indicated that the optimum medium components were: 0. 81% citric acid, 0. 35% NH4Cl and 0. 18% corn syrup. On those conditions, it was predicted that the highest production of carotenoid was 13. 34 mg/L, which was 2. 04 times higher than initial carotenoid yield before optimization.

OBJECTIVE: To observe the changes of malondialdehyde (MDA), superoxide dismutase (SOD), tumor necrosis factor-alpha (TNF-alpha), and interleukin-1beta (IL-1beta) in rat brain tissue and to explore the mechanism of secondary cerebral injury after brain concussion. METHODS: The brain concussion model was established with the pathological changes of rat brain tissue by Weil stain. The expressions of MDA and SOD in brain tissue were examined by photochemical method. The expressions of TNF-alpha and IL-1beta in cerebral cortex and hippocampus were examined by immunochemistry. RESULTS: Nerve myelin sheath showed disorder, disruption, gryposis and swelling by Weil stain. Above changes were more severe at 12h. The quantity of MDA in rat brain tissue after concussion was significantly higher than that in the control group. The activity of SOD was significantly lower than that in the control group. The expressions of TNF-alpha and IL-1beta increased more significantly in cerebral cortex and hippocampus in rat brain tissue after concussion than that in the control group. CONCLUSION Oxidative stress and inflammatory injury in the rat brain tissue, which may play an important role in secondary cerebral injury after concussion.
Animals ; Brain/metabolism* ; Brain Concussion/metabolism* ; Brain Injuries ; Hippocampus ; Interleukin-1beta/metabolism* ; Malondialdehyde/metabolism* ; Oxidative Stress ; Rats ; Superoxide Dismutase/metabolism* ; Tumor Necrosis Factor-alpha/metabolism*

OBJECTIVE: To investigate the role of endoplasmic reticulum stress (ERS) in lipopolysaccharide (LPS)-induced hepatocyte apoptosis. METHODS: Cells of the rat hepatocyte line BRL were cultured. The hepatocytes were treated with LPS, ERS inducer thapsigargin (TG), and ERS inhibitor 4-phenylbutyric acid (4-PBA), respectively or in their different combination. The cell viability was measured by MTT assay. The cyto-nuclear morphological changes of apoptosis cells were detected by the fluorescent dye Hoechst 33258. The apoptosis rate was assessed by flow cytometry with Annexin V-FITC/PI double-staining. Expressions of GRP78 as ERS marker protein, CHOP, caspase-12 and cleaved-caspase-3 as ERS related protein were detected by Western blotting. RESULTS: LPS could cause a decrease in cell viability and an increase in apoptosis rate in a dose- and time-dependent manner. The expression of GRP78, CHOP, caspase-12 and cleaved-caspase-3 proteins were significantly increased with LPS treatment. TG led to a marked decrease in cell viability and an increase in apoptosis rate, which aggravated the hepatocyte injury induced by LPS; whereas 4-PBA alleviated LPS-induced apoptosis. CONCLUSION ERS mediates LPS-induced hepatocyte injuries, indicating that ERS may play a vital role in the pathogenesis of LPS-induced hepatocyte injuries.
Animals ; Apoptosis ; Caspase 3 ; Cell Survival ; Endoplasmic Reticulum Chaperone BiP ; Endoplasmic Reticulum Stress ; Heat-Shock Proteins ; Hepatocytes ; Lipopolysaccharides ; Phenylbutyrates ; Rats

In practice of forensic medicine, potential disease can be associated with fatal asphyxia in restraint position. Research has demonstrated that nitric oxide (NO) and nitric oxide synthase (NOS) are plentifully distributed in skeletal muscle, contributing to the regulation of contractile and relaxation. In the current study, respiratory functions, indices of diaphragmatic biomechanical functions ex vivo, as well as NO levels in serum, the expressions of diaphragmatic inducible NOS (iNOS) mRNA, and the effects of L-NNA on contractility of the diaphragm were observed in sepsis induced by cecal ligation and puncture (CLP) under the condition of restraint position. The results showed that in the CLP12-18h rats, respiratory dysfunctions; indices of diaphragmatic biomechanical functions (Pt, +dT/dt(max), -dT/dt(max), CT, Po, force over the full range of the force-frequency relationship and fatigue resistance) declined progressively; the NO level in serum, and iNOS mRNA expression in the diaphragm increased progressively; force increased significantly at all stimulation frequencies after L-NNA pre-incubation. Restraint position 1 h in CLP12 h rats resulted in severe respiratory dysfunctions after relative stable respiratory functions, almost all the indices of diaphragmatic biomechanical functions declined further, whereas little change took place in NO level in serum and diaphragmatic iNOS mRNA expression; and the effects of L-NNA were lack of statistical significance compared with those of CLP12 h, but differed from CLP18 h group. These results suggest that restraint position and sepsis act together in a synergistic manner to aggravate the great reduction of diaphragmatic contractility via, at least in part, the negative modulation of NO, which may contribute to the pathogenesis of positional asphyxia.
Animals ; Asphyxia ; Diaphragm/physiology* ; Muscle Contraction ; Muscle, Skeletal ; Nitric Oxide/metabolism* ; Nitric Oxide Synthase ; Nitric Oxide Synthase Type II ; Rats ; Respiration Disorders ; Restraint, Physical ; Sepsis

OBJECTIVETo evaluate clinical results of early total care (ETC) treatment for elderly patients with intertrochanteric femur fractures.

METHODSClinical data of 106 elderly patients with intertrochanteric fracture treated from January 2012 and February 2015 were retrospectively studied. According to whether receiving the early total care mode, the patients were divided into 2 groups, 34 cases were diagnosed and treated with early total care pattern (ETC group), including 14 males and 20 females with an average age of (74.88 ± 4.38) years old ranging from 70 to 86. According to Evans types, 4 cases were type I, 5 cases were type II, 13 cases were type III, 11 cases were type IV, 1 case was type V. Seventy-two patients were treated with conventional trauma method (conventional group), including 35 males and 37 females with an average age of (74.46 ± 3.63) years old ranging from 70 to 85. According to Evans type, 8 cases were type I ,13 cases were type II, 25 cases were type III, 25 cases were type IV, and 1 case was type V. All fractures were treated with proximal femoral nails anti-rotation (PFNA). Operative time, hospital stays, leaving bed time, complications, cases of death at 1 year after operation, postoperative Harris score at 12 months were observed and compared.

RESULTSAll patients were followed up, the time of ETC group ranged from 9 to 18 months with an average of 13.29 ± 1.51, and the time in conventional group ranged from 12 to 16 months with an average 12.93 ± 1.15, while there was no significant difference between two groups in time of following-up (t = 1.368, P = 0.174). There was no significant meaning in cases of death between ETC group (2 cases) and conventional group (8 cases). Three cases occurred complications in ETC group, and 20 cases in conventional group,there was obvious meaning between two groups (χ² = 0.739, P = 0.318). Operative time,hospital stays,leaving bed time in ETC group respectively was (2.03 ± 0.67) d, (15.41 ± 2.87) d and (3.62 ± 0.74) d; while in conventional group respectively was (4.17 ± 1.59) d, (20.11 ± 4.24) d and (5.35 ± 1.22) d; there were significant differences between two groups in operative time, hospital stays, leaving bed time. Postoperative Harris scores at 12 months in ETC group was (82.32 ± 4.56), and (79.24 ± 5.52) in conventional group, there was obvious meaning between two groups (t = 2.833, P = 0.006).

CONCLUSIONETC pattern is a novel method for diagnosis and treatment of intertrochanteric femur fractures in elderly, it could shorten operative time, hospital stays, leaving bed time, decrease complications and promote recovery of function.

Aged ; Aged, 80 and over ; Female ; Femur ; injuries ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; Hip Fractures ; surgery ; Hip Joint ; surgery ; Hospitalization ; Humans ; Male ; Operative Time ; Postoperative Care ; Retrospective Studies

Objective To explore the effect of uhrasound-phoresis of Chinese herb in the treatment of frac- ture healing on rats.Methods The animal model of femur fracture was established in 36 rats,who were divided in- to 4 groups after operation.The rats in the ultrasound group were given ultrasound treatment daily.The herbal group was given Chinese herb applied on the fracture site.The experimental group was given uhrasound-phoresis of Chinese herb on the site of fracture.The control group was housed without any treatment.All rats were sacrificed at 30 days and the bony callus were harvested and observed with histological anti immunohistoehemical examination.Results The histological examination showed that the appearance of cartilaginous and bony callus in the experimental group were earlier than those in other groups,Immunohistochemical examination showed that the expression of collagenⅠin the experimental group was significantly higher than that in ultrasound group and herbal group(P

Objective To evaluate the long-term effect of hepatic arterial embolization with pingyangmycin-lipiodol emulsion (PLE) in patients with cavernous hemangiomas of the liver (CHL)and its influence factors.Methods One hundred and fifty-six hemangiomas that were diagnosed by imaging examinations or confirmed pathologically by surgery in 105 patients with integral follow-up data were analyzed retrospectively in this paper.All hemangiomas were divided into 4 groups according to their size (the largest size of hemangioma)by the authors as follows; A group(≤3 cm, n=25); B group(>3 cm and<5 cm, n =32) ;C group(≥5 cm and<10 cm, n=58); and D group(> 10 cm, n=41) .According to the number of abnormal sinusoids filled with the contrast medium in the angiography, 156 hemangiomas of this series were further classified as three types: abundant type (n=90) ; sparse type (n=9) and the middle type (n=57) .Hepatic arterial embolization with PLE was performed in the 105 patients with total 135 procedures (including single procedure in 75 patients, twice procedures in 30 patients).All of the 105 patients were followed up 3 to 8 years(mean 4 years) using US, CT or MRI or DSA.The criteria for the evaluation of curative effect were classified as best, good, recovery and no change in this series, and the sum of best and good effects were defined as the total effective rate.Then, the total effective rate of single procedure between the A, B groups and C, D groups; and the total effective rate of single procedure and twice procedures in the D groups, were compared statistically by X~2 test respectively.And the influence factors, included of the variant size of hemangiomas, abundant or sparse of abnormal sinusoids and the number of treatment procedures, were also analyzed and evaluated respectively.Results The total effective rate of the 156 hemangiomas in 105 patients was 92.95% (145/156).After single procedure of hepatic arterial embolization with PLE, the total effective rate of A and B groups were 100% (57/57), that of C and D groups were 85.86% (85/99), and the difference reach significant (X ~2=8.8553, P<0.01).In the D group, the total effective rate in group with twice procedures (100.00%,30/30) was significant higher (X~2 =5.2642,P<0.05) than those of group with single procedure (72.73%,8/11).Of this series, during the period of following-up, no recurrent or severe complications were observed.Conclusions A best long-term curative effect (complete cure) is usually obtained in the small hemangiomas with abundant abnormal sinusoids, and a satisfactory long-term curative effect can also be achieved in the larger or multiple hemangiomas, particularly in those hemangiomas with abundant abnormal sinusoids by using the repeat procedures of hepatic arterial embolization with PLE.

Objective To report the surgical method and treatment outecome of transfer of free upper limb lateral bone-skin flap for repair of bone defects and nonunion in hand and forearm.Methods 17 cases of hand composite trauma,5 cases of forearm composite trauma and 2 cases of nonunion in forearm were treated with free bone-skin flaps in distal humerus,whose pedicle was the posterior branch of radical collateral artery. Area of the flap was 2cm?3cm to 8cm?10cm,length of exseeted bone was 3～6 cm.Results All the bone-skin flaps completely survived,the donor area all achieved primary healing,On postoperative 1～4 years follow-up,the texture of the flap was excellent,and bone union was obtained in all transplants,the donor area of distal lateral humerus became thicker and thicker with new cortical bones formed.According to the upper arm function assessment criterion issued by hand surgery association of Chinese medical association.The hand function had excellent results in 17 cases and good results in 2 cases.Conclusion The bone-skin flap has following advantages:easy dissection,reliable blood supply,and no major vessel needed to be sacrificed,so it is an effective method for repair of skin and soft tissue with bone defects in hand and forearm,It.also can be used to repair refractory nonunion in ulna and radius.

Objective To investigate the carrying status and characteristics of Clostridium difficile isolated from infants.Methods Two hundred and thirty-eight stool specimens were collected from infant younger than 1 year old,that were hospitalized or outpatient from August to November 2015.Immunochromatography targeted GDH and toxin A&B of C.difficile was used for C.difficile screening,and those positive specimens were inoculated in CDIF and anaerobic culture.C.difficile isolates were genotyped by using slpA sequence typing (slpA ST),and tcdA,tcdB,cdtA and cdtB of C.difficile isolates were detected by PCR.Results Fifty C.difficile strains were isolated from 238 stool samples,and the isolated rates of C.difficile from <3 months,3 months to <6 months,and 6 months to 1 years old groups were 9.3%,17.6% and 27.3%(χ2=6.940,P=0.031<0.05),respectively.52.0%(26/50) of the C.difficile isolates were toxigenic,and 69.2% (18/26) toxigenic isolates harbored tcdA+tcdB+cdtA-cdtB-.Fifty C.difficile isolates were genotyped as 11 slpA STs,slpA ST fr-02 and kr-02 were the commonest genotypes in toxigenic C.difficile isolates;however,that was slpA ST xr-03 in non-toxigenic isolates.Conclusion High C.difficile carriage is found in infants younger than 1 year old,and more than half of C.difficile isolates are toxigenic.Most of toxigenic isolates harbored toxin A and B.The genotype of C.difficile isolates is different between toxigenic isolates and non-toxigenic isolates.

OBJECTIVE: To investigate effects of antioxidant stress protein heme oxygenase-1 (HO-1) on lipopolysaccharide (LPS)-induced endoplasmic reticulum stress (ERS) of rat hepatocytes. METHODS: The BRL cells (rat hepatocyte cell line) were cultured. The hepatocytes were treated with LPS, LPS+HO-1 siRNA, HO-1 siRNA and PBS solution, respectively. The cell viability was measured by trypan blue exclusion test. The apoptosis cells were detected by the fluorescent dye Hoechst 33258. Expressions of GRP78, CHOP, caspase-12 and HO-1 were detected by Western blotting. RESULTS: LPS caused an increase of HO-1 protein expression of rat hepatocytes in a dose-dependent and time-dependent manner, a up-regulation of GRP78, CHOP and caspase-12, a decrease in cell viability, and an increase in apoptosis rate of hepatocytes. Pretreatment of HO-1 siRNA inhibited the up-regulation of LPS-induced HO-1, however, aggravated ERS and cellular injury. CONCLUSION HO-1 inhibites ERS-mediated cellular injury of rat hepatocytes induced by LPS.
Animals ; Apoptosis/physiology* ; Endoplasmic Reticulum/metabolism* ; Endoplasmic Reticulum Stress/physiology* ; Heme Oxygenase-1/pharmacology* ; Hepatocytes/metabolism* ; Lipopolysaccharides/pharmacology* ; Rats