OBJECTIVE:To study the influencing factors for the extraction of the genomic DNA from Paeonia Suffruticosa.METHODS:Taking Paeonia Suffruticosa(root bark of Chinese medicinal herb) as material to investigate the influencing factors including concentrations of the NaCl and beta-mercaptoethanol,temperature and time of water bath,RNaseA,PCR(polymerase chain reaction) system etc in the buffer solution on the basis of modified CTAB method.RESULTS:The DNA obtained by modified CTAB method was pure,integrated,with the value of A260/A280 ranged from 1.8 to 2.0,the ampl-ified bands of PCR were clear and bright,which lay a solid foundation for the following molecular biology experiments.CONCLUSION:The modified CTAB method is economical,rapid and efficient,and it can be served as an extraction of genomic DNA from root bark Chinese medicinal herb as well as a theoretical basis for full scale production.

Objective:To investigate the effect of ultrasound-guided transverse abdominal plane block combined with dexmedetomidine on cesarean section.Methods:Eighty pregnant women undergoing elective cesarean section under general anesthesia in Binzhou Central Hospital from October 2017 to October 2019 were included in this study. They were randomly assigned to undergo either cesarean section under general anesthesia followed by patient- controlled intravenous analgesia (control group, n = 40) or ultrasound-guided transverse abdominal plane block combined with intravenous dexmedetomidine administration (observation group, n = 40). Analgesic efficacy and the analgesic effect on puerperas and their newborns were compared between the observation and control groups. Results:At 2, 4 and 12 hours after surgery, the Visual Analogue Scale score in the observation group was (2.1 ± 0.9) points, (2.2 ± 0.7) points, (3.0 ± 0.6) points, respectively, which was significantly lower than (4.4 ± 1.3) points, (4.9 ± 1.2) points, (3.9 ± 0.8) points in the control group, respectively ( t = 8.946, 7.854, 6.246, all P < 0.05). At 2, 4 and 12 hours after surgery, the comfort score in the observation group was (3.4 ± 0.8) points, (3.2 ± 0.7) points, (3.3 ± 0.7) points, respectively, which was significantly higher than (2.1 ± 0.7) points, (2.2 ± 0.9) points and (2.5 ± 0.8) points respectively in the control group ( t = 7.235, 6.130, 5.953, all P < 0.05). After surgery, the number of times the demand button of the anesthesia pump in the observation group was pressed was (8.4 ± 2.9), with the rate of patient satisfaction of 97.50% (39/40), which was significantly higher than that in the control group [13.1 ± 3.7, 77.50% (31/40), t = 7.036, χ2 = 9.347, both P < 0.05). At 1 and 2 days after surgery, the cumulative amount of anesthetics used for patient-controlled intravenous analgesia was (52.1 ± 1.9) mL and (121.7 ± 4.1) mL respectively, which were less than (63.6 ± 2.4) mL and (146.5 ± 5.3) mL in the control group, respectively ( t = 6.240 and 8.842, both P < 0.05). The score of extubation quality in the observation group was significantly lower than that in the control group [(1.3 ± 0.3) points vs. (3.2 ± 0.5) points, t = 6.664, P < 0.05]. There was statistically significant difference in cortisol content between the two groups at 10 minutes after extubation and at the time points of fetus removal, skin incision and intubation (all P < 0.05). In the observation group, the partial pressure of carbon dioxide in the umbilical cord blood was significantly reduced, the partial pressure of oxygen and the oxygenation index in the umbilical cord blood were significantly increased, and Apgar score at 1 minute after birth was significantly increased (all P < 0.05). Conclusion:Ultrasound-guided transverse abdominal plane block combined with dexmedetomidine can enhance the anesthetic effect on cesarean section, prolong analgesic time, reduce the amount of analgesics, improve the operation effect, reduce maternal stress responses, and improve maternal and infant health.

0.05). It was established that both TMP and LPS had less cytotoxic property in this condition. LPS could inhibit the NOS activity (P

A secretive expression vector of Pichia pastoris system which can be used for the direct cloning of PCR products was constructed,and was verified through the expression of recombinant cellobiohydrolase II in Pichia pastoris.A randomly selected fragment was amplified with properly designed primers by PCR.The XhoI and Eam1105Ⅰ restriction sites were included in the 5'end of the fragment,and the Eam1105Ⅰ and XbaI restriction sites were included in its 3'end.The PCR amplified product was inserted into the P.pastoris expression plasmid pPICZ?A through XhoI and XbaI restriction sites and the resultant plasmid was digested with Eam1105Ⅰ,and lastly the big fragment was recovered,generating the P.pastoris expressive Tvector pPICZ?T.Then the cellobiohydrolase II of T.reesei was successfully expressed in P.pastoris with this expressive Tvector.Such results indicated that the constructed expression Tvector was convenient for PCR product cloning,and was effective for heterologous protein expression in P.pastoris.On the other hand,the application of the expression Tvector avoided the introduction of additional amino acids at the Nterminus of the expressed protein,which generally occurred when normal expression vectors were used in secretive expression system.

Ear, Middle ; surgery ; Humans ; Otologic Surgical Procedures

The concentrations of berberine (BBR) and 8-cetylberberine (8-BBR-C16) in rat plasma and tissue were determined by RP-HPLC. Both the plasma pharmacokinetics characteristic and tissue distribution differences of BBR and 8-BBR-C16 were compared to provide experimental data for the mechanism research and further drug development. After the oral administrations of BBR and 8-BBR-C16 at the dose of 80 mg x kg(-1) for rats, the pharmacokinetics result showed that compared with BBR, the C(max) and AUC(0-t), of 8-BBR-C16 increased by 2.8 times and 12.9 times respectively, t1/2 extended from 3.61 h to 11.90 h. The tissue distribution result showed that compared with BBR, the concentration of 8-BBR-C16 in various organizations increased and the retention time extended remarkably. The maximum concentration was achieved in lung and the highest concentration in it was 3 731.82 ng x g(-1). After being derived, the C(max) in plasma and bioavailability of 8-BBR-C16 increased remarkably and the circulation time in vivo extended. The drug concentration in tissue increased remarkably, and the distribution ratio changed too, with strong targeting selection in lung.
Administration, Oral ; Animals ; Berberine ; analogs & derivatives ; pharmacokinetics ; Biological Availability ; Chromatography, High Pressure Liquid ; Rats ; Tissue Distribution

Objective To analyze and evaluate the effects of the tissue engineering bladder reconstruction on the upper urinary tract structure and function.Methods The 8 male beagles were randomly divided into the two groups:sham-operation group (group A,n=4)and the collagen scaffold repair group (group B,n=4).The bladder defect animal model was established in the group B by using the collagen scaffold materials to repair the bladder.The renal function related biochemical indicators were detec-ted and the renal Doppler ultrasonic examination was performed in each group before repair and in 23 weeks after repair.The speci-mens from the two groups were performed the gross morphology observation and the histology examination on postoperative 24 weeks.Results The renal Doppler ultrasound examination showed the normal kidney morphology and normal blood flow signal.In the general observation,no calculi and neoplasm were found in the kidney and ureter of the experimental dogs.The renal function related biochemical indicators had no statistically significant differences between the two groups(P>0.05).The histological exami-nation indicated that the organization structure was integrity,the nephrons in each group had no obvious pathological changes.Con-clusion Using the collagen scaffold materials to reconstruct the canine bladder has no adverse influence on the upper urinary tract structure and function,this tissue engineering approach has good feasibility.

Objective To explore the characteristics of T-cell receptor beta (TCRβ) gene rearrangement in children with T-cell acute lymphoblastic leukemia (T-ALL) and establish a system for quantitative detection of minimal residual disease (MRD) by real-time quantitative PCR (RQ-PCR) targeting the TCRβ gene rearrangement. Methods Multiplex PCR designed by the BIOMED-2 was used to detect TCRβ gene rearrangement in the bone marrow samples of 26 children with T-ALL. Sequences of junctional region were then compared and analyzed in IMGT database. Allele specific oligonucleotide (ASO) upstream primers were designed complementary to the V-D-J or D-J junctional region of TCRβ gene rearrangements. Samples at diagnosis were serially diluted in DNA obtained from mononuclear cells (MNC) from a pool of 20 healthy donors to generate the patient-specific standard curves. Subsequently, TCRβ RQ-PCR was applied to six patients to quantify MRD with germline Jβ primer/probe combinations. To determine the quantity and quality of DNA, we also used RQ-PCR for the N-ras gene.Results Clonal rearrangements were identified in 92.3% of the children with T-ALL ( Vβ-Dβ-Jβ rearrangements in 84.6% and Dβ-Jβ rearrangements in 50% ). Comparative sequence analysis of 42 TCRβ recombination revealed that two downstream Vβ families (BV5, BV6) were preferentially used. The segment Jβ2. 7 was dominant in childhood T-ALL. Jβ1. 3, Jβ2.4, and Jβ2.6 were not detected. The slope of the standard curves was from - 3.54 to -3.37 with the intercepts between 19.35 and 20.51. The correlation coefficients of all the 6 standard curves were ≥0.98. None of the cases had a quantitative range of RQ-PCR lower than 10<'-4>. During the follow-up, an increased incidence of MRD was found before relapse. Conclusions RQ-PCR, which is a highly sensitive and specific method for detection of TCRβ gene rearrangements, will be of high value to study MRD in T-ALL. Close monitoring of MRD is of great importance for prognosis and follow-up of the patients with the disease.

OBJECTIVETo observe the clinical effect and safety of circumcision stapler in the treatment of phimosis and redundant prepuce.

METHODSWe treated 120 patients with redundant prepuce or phimosis using circumcision stapler and another 60 by conventional dorsal-incision circumcision. We observed intraoperative blood loss, operation time, postoperative pain, wound healing time, cosmetic appearance of the penis, and postoperative complications and compared them between the two groups of patients.

RESULTSStapler circumcision showed obvious advantages over the conventional method in intraoperative blood loss ([2. 3 ± 1. 3] vs [15.6 ± 2.9] ml), operation time ([7.1 ± 1.4] vs [22.6 ± 4.6] min), wound healing time ([12.0 ± 2.9] as [16.3 ± 3. 1] d), postoperative pain score (1. 9 ± 1. 3 vs 5. 2 ± 1. 7), incision edema, and cosmetic appearance of the penis (all P <0. 05). Besides, stapler circumcision exempted the patients from stitch-removal pain. However, the incidence rate of postoperative local ecchymosis was significantly higher in the circumcision stapler group than in the conventional circumcision group (20. 8% vs 8. 3% , P <0. 05).

CONCLUSIONCircumcision stapler, with its advantages of easier manipulation, shorter operation time, better cosmetic penile appearance, less pain, and fewer complications, is superior to conventional circumcision in the treatment of phimosis and redundant prepuce.

Blood Loss, Surgical ; Circumcision, Male ; instrumentation ; methods ; Ecchymosis ; etiology ; Humans ; Male ; Pain, Postoperative ; Penis ; abnormalities ; Phimosis ; therapy ; Postoperative Complications ; Postoperative Period ; Surgical Staplers ; adverse effects ; Wound Healing

OBJECTIVETo investigate the hemodynamics evidence of the descending branch of lateral circumflex femoral artery in a reversed way. To explore the clinical result of using the reversed descending branch of the lateral circumflex femoral artery as the receipt artery for free flaps for reconstruction of the leg soft-tissue defect.

METHODSFrom October 2005 to February 2012, 38 patients with severe leg soft-tissue defects were treated. The proximal antegrade and retrograde mean artery pressure of the descending branch of the lateral circumflex femoral artery in 16 of 38 patients were recorded during operation. All wounds had osteomyelitis, bone and tendon exposure requiring coverage reconstruction. And there was no recipient artery in the injured lower leg for free flaps in all 38 patients. Reversed descending branches of lateral femoral circumflex arteries were used as recipient arteries for free flaps (free latissimus dorsi flap, free thoracoumbilical flap, and free anterolateral thigh flap) in all patients. The flap donor site was closed directly or with the skin graft.

RESULTSThe proximal antegrade mean artery pressure of the descending branch of lateral circumflex femoral artery was(81.6 +/- 12.4) mmHg. The proximal retrograde pressure was(48.2 +/- 10.7) mmHg. The proximal retrograde mean artery pressure was 59.07 percent of the proximal antegrade pressure. The donor skin graft survived and wound healed primarily. After operation, 2 flaps had distal partial necrosis and healing was achieved after dressing change. All the other flaps survived completely without vascular problems. All the patients were followed up for 11 months to 2.5 years (mean, 1.6 years). The flap appearance was satisfactory. The texture and color of flaps in all cases were good.

CONCLUSIONSThe reverse descending branch of lateral circumflex femoral artery is a reliable recipient artery for the free flaps. It is an easy and simple technique that can be used for reconstruction of the defects in the lower leg, with the reversed descending branch of lateral circumflex femoral artery as recipient artery.

Adolescent ; Adult ; Blood Pressure ; Female ; Femoral Artery ; physiopathology ; surgery ; Free Tissue Flaps ; blood supply ; Hemodynamics ; Humans ; Lower Extremity ; injuries ; Male ; Middle Aged ; Soft Tissue Injuries ; surgery ; Young Adult