Objective To develop a rapid,sensitive,specific and accurate quantitative duplex real-time PCR assay for detection of Listeria monocytogenes and Shigella.Methods Two sets of specific primers and probes were selected according to Listeria monocytogencs hly gene and Shigella ipaH gene.The target hly and iPaH fragments were amplified by PCR,and used to construct recombinant pGEM-T-hly and pGEM-T-ipaH respectively.The two recombinant circular plasmid DNAs were linearized with EcoR I that did not cut within the target DNA fragment.The ten-fold dilutions of plasmid were subjected to the standard quantitation curve in duplex real-time PCR assay.Various genomic DNAs of Listeria innocua,Listeria weshimeri,Salmonella,Staphylococcus aureus,Bacillus subtilis,Escherichia coli and Proteus were used as negative controls to confirm the specificity of duplex real-time PCR assay.The assay was also used to detect Listeria monocytogenes and Shigella in artificially contaminated sterilized skim milk.Results The recombinant plasmids were constructed successfully,hly probe(rAM and TAMRA double labelled)and ipaH probe (HEX and TAMRA double labelled)were used to develop an optimized PCR successfuliv.Conclusion The selected primers and probes showed high specificity for these two target bacteria,the linear range of the assay was good(105-101 copies/μl,R2≥0.998)and sensitivity Was 10 copies/PCR.Following a DNA extraction method which combined EZ Spin Colum Genomic DNA Isolation Kit(BBI)/Phenol-chloroform,the sensitivity of assay Was 102CFU/ml for both Listeria monocytogenes and Shigella in artificially contaminated sterilized skim milk,which equivalents to 10 CFU/PCR.

[Objective] To analyze the constituents of volatile oil from Nervilla fordii (NF) by gas chromatography-mass spectrometry (GM-MS). [Methods] Volatile oil was extracted from NF by steam distillation and the constituents of volatile oil were isolated and identified by GM-MS. The relative contents of the volatile constituents were calculated by ionic flow chromatographic area normalization method. [Results] Eighty-nine components were separated. Among them, fifty-three components were identified, accounting 91.82% of the total volatile oil. [Conclusion] The main constituents identified from NF were: 2-pentadecanone, 6,10,14-trimethyl (13.55%), 4-ethyl-trans-3-thiobicyclo [4,4,0] decane (6.54%), benzenesulfonamide, 4-methyl-N- (2-oxo-2 phenylethyl) (6.33%), phytol (6.32%),?-carinol (4.54%), ?-ionone (4.43%), caryophyllene oxide (4.13%). The amount of the above constituents accounts 45.84% of the total volatile oil.

Animals ; Cholesterol ; blood ; Hyperlipidemias ; blood ; drug therapy ; Leptin ; pharmacology ; therapeutic use ; Male ; Mice ; Mice, Inbred Strains

Actins ; metabolism ; Bone Neoplasms ; metabolism ; pathology ; surgery ; Desmin ; metabolism ; Diagnosis, Differential ; Fibrosarcoma ; metabolism ; pathology ; Humans ; Leiomyosarcoma ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Neurilemmoma ; metabolism ; pathology ; Tibia ; Vimentin ; metabolism

OBJECTIVETo investigate the effect of different pH on rectum permeability of chlorogenic acid and geniposide.

METHODFour kinds of Reduning suppositories of different pH were separated and put into the rectum to study the suppositories in vitro and the content of chlorogenic acid and geniposide samples was determined by HPLC to calculate the permeation in 24 hours.

RESULTWith increase of pH within 2.5-7.4, the steady state flux of chlorogenic acid was increased, but the steady state flux of geniposidesamples was steady.

CONCLUSIONAdjusted the pH can increase the rectum permeability of active ingredients in Reduning auppositories.

Animals ; Chlorogenic Acid ; pharmacokinetics ; Drugs, Chinese Herbal ; pharmacokinetics ; Hydrogen-Ion Concentration ; Iridoids ; pharmacokinetics ; Male ; Permeability ; Rats ; Rats, Sprague-Dawley ; Rectum ; metabolism ; Suppositories ; pharmacokinetics

Objective Study on the absorption,distribution,transformation,excretion and toxicity of four forms of arsenic in rats.Methods Four thousand and fifty data were obtained from 28 days animal metabolism experiments of 81 rats exposed to the four As species.Distribution,metabolism,excretion,and subacute toxicity of 4 As species were compared by analyzing the arsenate [As(V)],arsenite [As(Ⅲ)],methyl arsenate (MMAV) and dimethyl arsenate (DMAV) content of rat feces (excrement and urine),blood,seven organs,and liver and kidney pathology.Results After oral administration of each As species,82.9％ of As(Ⅲ),85.1％ of As(V),95.0％ of MMAV and 96.2％ of DMAV were accumulatively secreted via feces and urine,while 16.2％ of As(Ⅲ),14.1％ of As(V),4.65％ of MMAV and 0.120％ of DMAV were detected in blood.The DMAV accumulated in blood and urine after dosing As(Ⅲ),As(V) and MMAV,and the content of DMAV in the blood was 8 times greater than urine.Four kinds of arsenic had effects on rat liver and kidney function,the influence on liver was acute phase effect while that on renal was cumulative effects.The effect of As(Ⅲ) was the most significant,followed by MMAV.Four kinds of arsenic could cause tissue deformation and inflammatory cells infiltration,but showed no difference between groups.Conclusion The distribution,metabolism,excretion,and toxicity of the four kinds of arsenic in rats were different in varying degrees.The research of health risk for arsenic should consider its different forms.

Objective To investigate the rat bone structural changes of lower limb following lumbar nerve dorsal roots section.Methods Forty-eight mature female Wistar rats were divided into posterior radi- cotomy(PR)and comtrol groups randomly.The bilateral femoral bone mineral density(BMD)and biome- chanics characteristics were analyzed 2,4 and 8 weeks after the radicotomy.The same operation except the radicotomy was done in the sham group.Results In PR group,2,4,and 8 weeks after the radicotomy,the BMD of femur was(0.221?0.008)g/cm~3,(0.213?0.015)g/cm~3 ,and(0.216?0.105)g/cm~3 ,respective- ly;while that was(0.223?0.005)g/cm~3,(0.218?0.014)g/cm~3 ,and(0.208?0.111)g/cm~3 in control group.No significant difference was observed between the two groups(P＞0.05).In PR group,2,4,and 8 weeks after the operation,the mean maximum load in three-point bending test of femun midshaft was(93.64?8.76)N,(89.77?11.18)N and(93.21?8.74)N,respectively,and was lower than the values of the con- trol group at the same time point(95.94?6.29)N,(91.63?9.43)N,(95.57?8.64)N,However,there was no significant difference between the two groups(P＞0.05).Accordingly,there was no significant difference in the energy absorption in femun midshaft between the two groups(P＞0.05).Conclusion The selective rhizotomies of part lumbar never dorsal roots might not cause the loss of the femur BMD and the change of bio- mechanics property significantly in short period.

OBJECTIVE: To investigate the clinical effect of a modified arthroscopic outside-in suture technique in the treatment of meniscus tear using a spinal needle. METHODS: From January 2015 to October 2017, 95 patients treated with this method were followed-up. Among these cases, there were 36 males and 59 females. The age of the patients ranged from 16 to 77 years, (46.79±18.07) years in average. Among them, there were 28 patieats aged 16-35, 53 patients aged 36-65, and 14 patients aged over 65 years old. 28 cases were diagnosed with medial meniscus tear, 43 cases with lateral meniscus tear and 24 cases with both medial and lateral meniscus tear. Causes of the injury included sports, sprain, etc. According to Barrett standard, the clinical healing of meniscus tear was judged. Lysholm score, knee range of motion, visual simulation score (VAS) and magnetic resonance imaging (MRI) were used to evaluate the postoperative knee function and recovery of the patients. RESULTS: The 95 patients were followed up for 22 to 36 months, with an average of (28.32±3.98) months. According to Barrett standard, 90 patients (94.7%) obtained meniscus clinical healing. Meniscal healing rates were 96.43%, 96.23% and 85.71% in the three age groups, respectively. The meniscal healing rate was lower in the elderly group, but there was no significant difference in statistical results (P=0.262). Five patients had deep tenderness in the joint space of the injured side, and the overstretch test was positive. The preoperative and postoperative VAS scores, Lysholm scores and knee motion were compared in each group, and the differences were statistically significant (P < 0.01). At the end of the last follow-up, there were no cases of knee joint effusion, swelling and interlocking, and the joint function was effectively improved in most patients. No surgical site infection, periarticular vascular/nerve injury or knotting reaction was found during the follow-up. CONCLUSION This modified arthroscopic outside-in suture technique using a spinal needle has the characteristics of simple operation, small trauma and rapid recovery, and the mid-term follow-up results were satisfactory. Therefore, we consider this method to be a safe and efficient method for the treatment of meniscus anterior horn and body tear.
Aged ; Arthroscopy ; Female ; Follow-Up Studies ; Humans ; Knee Injuries/surgery* ; Male ; Punctures ; Suture Techniques ; Tibial Meniscus Injuries/surgery*

OBJECTIVE: To evaluate the surgical technique and clinical effect of less invasive intrame-dullary nail fixation combined with titanium cable cerclage in the treatment of subtrochanteric fractures. METHODS: A retrospective study was performed in 46 cases of subtrochanteric fractures in Peking University People's Hospital from January 2015 to December 2017. Among them, there were 14 males and 32 females, with an average age of (77.83±10.66) years (44-92 years); 17 cases on the left side and 29 cases on the right side. The causes of injury included crash from a height, traffic accident and accidental fall. According to Seinsheimer classification, there were 26 cases of type Ⅱ, 11 cases of type Ⅲ, 9 cases of type Ⅳ, and these cases were all closed injury. After admission, these patients underwent continuous tibial tuberosity bone traction to maintain the length and force line of the lower extremity, so as to reduce the difficulty of intraoperative fracture reduction. Anticoagulant therapy was given before operation to reduce perioperative thrombotic complications. All the patients were treated with less invasive intramedullary nail fixation combined with titanium cable cerclage. Operation time, blood loss during surgery, time of fracture healing were recorded, Harris and Sanders scoring system were used to assess hip function after operation at each follow-up time point. RESULTS: All the included patients underwent surgery successfully. Average operative time and intraoperative blood loss of these patients were (131.09 ± 20.06) min and (191.96±111.03) mL, respectively. All the patients were followed up satisfactorily, with an average follow-up time of 28 months. The fractures received bone healing within 3-6 months, average hospital stay was (10.61±2.85) days. The Sanders score was excellent in 3 cases, good in 37 cases and common in 6 cases, with an excellent and good rate of 86.96%. The Harris score was excellent in 6 cases, good in 36 cases, with an excellent and good rate of 91.30%. There were no cases of wound infection, loss of reduction, nonunion of fracture or internal fixation failure. Hip pain symptoms were effectively relieved in most patients. CONCLUSION Less invasive intramedullary nail fixation combined with titanium cable cerclage can obtain good alignment and stability of fracture ends, which is an effective method for the treatment of subtrochanteric fractures.
Aged ; Aged, 80 and over ; Bone Nails ; Female ; Fracture Fixation, Intramedullary ; Fracture Healing ; Hip Fractures/surgery* ; Humans ; Male ; Retrospective Studies ; Titanium ; Treatment Outcome

In this paper, the five strains of Polygonatum odoratum were used as the experimental materials to test the supercooling point, freezing point, the degree of supercooling, the transition stage time, cooling time and water composition of the plant tissue. The cold resistance of P. odoratum was analyzed with the Gray Correlation Method. The results showed that the cold resistances of the five strains of P. odoratum were different, and the water content of plant tissue had some relevance with freezing point and supercooling point, whereas, it could not be measured when the moisture content was too low. The order of cold resistance of the five strains of P. odoratum was ZJCY, DYYZ, XYYZ, CYYZ and JZ I.
Cold Temperature ; Plant Roots ; chemistry ; physiology ; Polygonatum ; chemistry ; classification ; physiology ; Water ; analysis