DNA extraction is a basic technology of molecular biology. The purity and the integrality of DNA structure are necessary for different experiments of gene engineering. In recent years there have been some new or improved DNA extraction methods appeared. The methods of DNA extraction from animals, plants, microorganisms and marine organisms were summarized in this article.

Objective To investigate the effect of exogenous murine interleukin-18 (IL-18) on early mouse collagen Ⅱ -induced arthritis (CIA). Methods Mice were injected intraperitoneallg IL-18 (0.2 μg/d) combination with IL-10 (0.1 μg/d), IL-4 (0.1 μg/d) and IL-12 (0.1 μg/d) daily for five days before the onset of CIA. The arthritis response was monitored visually by macroscopic scoring. Reverse transcription -polymerase chain reaction (RT-PCR) was employed to determine the mRNA expression of cytokine in patella with adjacent synovium in CIA mouse. Histology of knee was examined to assess the occurrence of cartilage destruction and bone erosion. Wilcoxon rank test was selected. Results IL-18/IL-4 treatment could slightly suppress the macroscopic score of arthritis, but a more pronounced amelioration was found in mice treated with the combination of IL-18 and IL-10 during early treatment. On 38 days after immunizatian macroscopic score in treated group (0.12±0.20) was significantly improves than in the control group (0.29±0.19, P<0.05). This resulted in both the suppression of macroscopic signs of inflammation and the reduction of cellular infiltrates in the synovial tissue, which provided the protection against cartilage destruction. Moreover, the expression of Thl cytokines [IL-18 (0.22±0.06), IL-12 (0.14±0.05)] and inflammatory cytokine [IL-6 (0.22±0.11)] was greatly inhibited both in the synovial tissue and in the articular cartilage in the treatment groups compared with those in the control groups (P<0.05). However, the mRNA levels of Th2 cytokines [IL-10 (6.35±0.12), IL-4 (3.57±0.13)] were up-regulated after IL-18/IL-10 treatment (P<0.05). Moreover, IL-18R (0.40±0.15) levels were down-regulated compared with those in the control group (P<0.05). T-bet mRNA levels were decreased in IL-18/IL-10 compared with the control group, and GATA-3 mRNA (5.71±0.11) levels were significantly higher than those in the control group (P<0.05). Conclusion Low dose IL-18/IL-10 treatment can inhibit Th1 cytokines expression and induce Th2 cytokines expression, which may be mediated not only by inhibiting Th1 responses through IL-18/IL-18R mechanism, but also by inducing anti-inflammatory mediators such as IL-10 and IL-4 through a GATA-3-dependent mechanism.

Objective To investigate the effect of exogenous murine interlukin-18 (mIL-18) on early and established murine coUagen-induced arthritis (CIA). Methods Mice were injected intraperitoneally with IL-18 (0.2 μg/mouse) daily for 5 days before or after the onset of CIA. The response was monitored visually by macroscopic scoring. Reverse transcription-polymerase chain reaction (RT-PCR) was employed to determine the mRNA expression of cytokines in patella with adjacent synovium in CIA mouse. Histolpgy of knee synovium was used to assess the occurrence of cartilage destruction and bone erosions. Results IL-18 alone had no effect on macroscopic score, occurrence of arthritis, advancement of histology on early stage of CIA. Moreover, expression of Th 1 cytokines and Th2 cytokines in the synovial tissue and" articular cartilage remained unchanged compared with the control group, however, a pronounced progression of histology was found in mice treated with IL-18 in estabhshed CIA. Forty-three days after immunization, the macroscopic score in the treated group (0.33±0.11 ) was significantly improved than in the control group (0.25±0.09) (P<0.05). Moreover, the mRNA levels of IL-10 and IL-18BP both in the synovial tissue and in the articular cartilage in the treated groups decreased significantly than those in the control groups (P<0.05). Conclusion Low dose mIL-18 alone has no effect on early stage CIA. But pronounced exacerbation is found in mice treated with IL-18 on established arthritis, which supports that IL-18 initiates this effect by inhibiting IL-10 and IL-18 BP.

Objective To investigate the expression of GR? mRNA in the peripheral blood mononuclear cells (PBMCs) of SLE patients and analyze the relationship between GR? and immune parameters, SLEDAI score, in order to reveal the role of GR? in the pathogenesis of SLE. Methods Applying reverse transcription-polymerase chain reaction(RT-PCR) technique to semiquantitatively analyze GR? mRNA expression in SLE patients and healthy volunteer individuals. The relationship between GR? and ANA, anti-ds-DNA antibodies, SLEDAI score was analyzed. Results The expression of GR? mRNA in SLE was significantly lower than that of control group (P

Objective To observe effect of Guben Xiaoliu capsule combined with FOLFOX4 regimen in treating advanced colorect cancer patients. Methods 78 advanced colorect cancer patients were randomly assigned to treatment group (38 patients) and control group (40 patients). Oxaliplatin 85 mg/m2 iv infusion for 2 hours,d1. CF 200 mg/m2 iv infusion for 2 hours followed by 5-FU 400 mg/m2 iv infusion for 22 hours,d1-2. Every two weeks was a cycle. The control group was treated by FOLFOX4 regimen,while Guben Xiaoliu capsule was added in the treatment group. Patients were evaluated after 4 cycles. Results Clinical beneficial rate (CR+PR+SD) of treatment and contral group were 76.3% and 57.5% respectively (P

Objective To study the characteristics of toe-to-thumb transplantation in a group of infants who completely lacked the thumb.The mean age was 24.6 months,and the youngest was 10 months old,The operative technique and rehabilitation were studied.Methods In two infants with Ⅲ-Ⅳ? defects of the thumb together with multiple injuries of fingers,the wounds were debrided and toe-to-thumb transplantation was done later.In case 3 Ⅲ? defect of the thumb together with palm injury,one-stage toe-to-thumb transplantation and the palm injury was covered with an abdominal flap were done at the same time.Considering the characteristic physiological structure of a dorsalis pedis and the first metatarsal arteries.In the infant,minimally invasive operation under 10? microscope was employed.A series of early functional exercises was given for the reconstructed thumbs.Results The reconstructed thumbs completely survived,hand function was recovered 3-6 months after the operation.With follow-up for longer than 2 years in 2 cases,it was noted that the reconstructed thumb grew consistent with the wounded hand.The appearance and function thumb were satisfactory.Conclusion Toe to thumb transference was successful in infancy,with the combined technique of minimally invasive and plastic surgery.Reconstruction of the thumbs can be completed in two stages on immediately after injury.The appearance and function can be restored.[

Objective To establish an effective technique in reservation and function reconstruction of a leg, in which all the muscles except gastrocnemius necrosed induced by the delayed necrosis of all the intermuscular vessels, the foot was linked only by skin, fascia and tibia. Methods The leg was linked with foot only by the skin, fascia, caput mediale of gastrocnemius and tibia when all the muscles, intermuscular vessels and fibula were removed. The direct island flap of tibialis posterior artery was cut from the healthy limb and used to cover the 4/5 length and 1/3 circumference of bone-fascia raw surface, then to anastomose the both arteries of the flap and the caput lateral gastrocnemius when dividing the pedicles. As soon as the raw surface healed, fuse the ankle in the functional situation and fix the flexor and extensor tendon in the proper situation. Results With sufficient blood supply from the skin and fascia of the flap, the raw surface healed soon, the severe ischemia pain relieved, and the circulation and sensation recovered gradually. The crutching and walking function of the leg reconstructed at last. Conclusion When all the necrotic intermuscular vessels and muscles except caput mediale of gastrocnemius were removed, the necrosis may be delayed by means of reticular collateral circulation among skin, deep and superficial fascia and tibia of the injured limb. However, the blood supply is very little, the severe ischemia wound and ischemia pain can't be healed. Through the tibialis posterior artery direct island skin flap of the healthy leg to cover the dystrophic wounds, the blood supply of the leg can be improved. As a result, the leg survives and its function is reconstructed, and an eternal axial-pattern artery for the wound leg through arteries anastomosis is rebuilt.

Objective To probe the pathogenesis of long bone defects and damage to epiphysis as the result of postoperative infections following fractures in the childhood of patients, and to look for ideal ways to prevent and repair the defects and damages. Methods The treatment of the infected bone defect of a tibia had been continued for one year following fracture in one four-year old patient, and the treatment of the infected bone defect of a humerus had been continued for ten years following fracture in another seven-year old patient, but all the treatments had failed. The injured limbs developed bone infection, bone defect, bone exposure, shortening, and pseudoarthrosis deformities at last. Then the lesions were completely cleaned, and the deformities of the infected bones were corrected in one stage, with reconstruction of the bone and the repair of the wounds by transplantations of fibular free flaps. Results Both fibular free flaps survived in one stage transplantation. The bones of the injured limbs healed half a year after the operations, and weight-bearing and walking functions of the injured limbs recovered as well. Conclusion Transplantation of fibular free flap was an ideal way to treat a long bone defect as a result of postoperative infection following fracture in the childhood. The early appropriate treatment of postoperative infection following fracture is very important to avoid bone and epiphysis damages.

Objective To investigate cause,diagnosis,treatment and prevention of vertebral artery injury in cervical spine surgery.Methods Data of 7 patients with vertebral artery injury caused by cervical spine surgery from October 2002 to April 2012 were retrospectively analyzed.There were 6 males and 1 female,aged from 23 to 65 years (average,48.9 years).The reasons of cervical spine surgery were as follows:cervical spondylotic myelopathy (5 cases),traumatic subluxation of C4 and C5 (1 case),and cervical spinal stenosis due to skeletal fluorosis (1 case).All cases had unilateral vertebral artery injury,including 4 cases in the left side and 3 cases in the right side.The cause,treatment and prognosis of vertebral artery injury in 7 patients were analyzed.Results Four patients suffered from vertebral artery injury during anterior cervical spine surgery; 2 cases were caused by excessive lateral trepanation,1 by excessive lateral discectomy using a curette,and 1 by Kerrison punch during decompression in a patient with anomalous vertebral artery due to traumatic subluxation of C4 and C5.Three patients suffered from vertebral artery injury during posterior surgery; 2 cases were caused by a drill bit during drilling lateral mass of C4 and 1 by a rongeur during removing posterior C1 arch in a patient with cervical spinal stenosis due to skeletal fluorosis.After vertebral artery injury,all patients were immediately treated by hemostatic tamponade,and then the incisions were closed.However,1 patient occurred delayed cervical hemorrhage 4 weeks after operation.Then he underwent vertebral artery angiography,balloon embolization and evacuation of hematoma.All cases had no cerebral infarction.However,two of them had postoperative temporary dizziness.Conclusion Vertebral artery injury in cervical spine surgery is a rare but serious complication,which is associated with surgical mistakes and anatomical variations.Hemostatic tamponade and embolization are effective for this complication.

The human proinsulin gene was introduced a cleanage site for furin with two point mutation by polymerase chain reaction. The mutated gene was subcloned to the retroviral vector, and transferred into HepG2 hepatocarcinoma cells. Mature insulin can be expressed and secreted into supernatant by transferred HepG2 cells. The site mutation method is concise and with no misproduction.