BACKGROUND:Keloid is the outcome of wound-healing process,and the result of massive accumulation of life-prolonged fibroblasts with gene mutation as well as the excessive synthesis of collagenous fibers.OBJECTIVE:To probe the relationship between the fibroblasts and the mutations of the exon-8 of Fas gene in keloid.DESIGN:An open study with gene sequence as the subjects of observation.SETTING :The Department of Plastic Surgery of Southern Hospital of the First Military Medical University.PARTICIPANTS:This experiment was carried out at the Tropical Disease Research Institute of the First Military Medical University of Chinese PLA in 2001. All keloid and hypertrophic scar tissues were obtained from the patients who received orthopedic surgical operations at the Southern Hospital, including 15 patients with keloid whose pathological areas were located respectively at the earlobe and the prothorax and 12patients with hypertrophic scars whose pathological areas being located at the instep and the elbow. At the same time, normal skin and the peripheral blood samples from the patients themselves with keloid were taken as the self-control and the skin and the peripheral blood samples from the normal people and the patients with hypertrophic scars were taken as the normal control.METHODS: PCR-SCCM technique and gene sequence analysis were used to detect the gene structure of exon-8 in the Fas gene from 15 patients.MAIN OUTCOME MEASURES:The gene structure of exon-8 in the Fas gene derived from the tissues and the peripheral blood samples of all the groups.RESULTS: ① Heterozygous loss was observed in the exon-8 of the Fas gene in all 15 keloid patients; ② Gene sequence was found to be abnormal in 11 cases out of 15 keloid patients, presenting gene mutation in 4 loci.CONCLUSION: Heterozygous loss and gene mutation was detected in the exon 8 of Fas gene of keloid, suggesting that Fas protein in keloid has functional defect that is closely associated with gene mutation.

BACKGROUND:Keloid is the outcome of wound-healing process,and the result of massive accumulation of life-prolonged fibroblasts with gene mutation as well as the excessive synthesis of collagenous fibers.OBJECTIVE:To probe into the structural relations of exons 7-9 of fibroblastic Fas gene in keloid tissues.DESIGN:A self-controlled experimental study with cicatricial tissues as the subjects.SETTING:Department of Plastic Surgery of Southern Hospital of the First Military Medical University of Chinese PLA.PARTICIPANTS:This experiment was carried out at the Tropical Disease Research Institute of the First Military Medical University of Chinese PLA in 2001. All keloid and hypertrophic scar tissues were obtained from the patients who received orthopedic surgical operations at the Southern Hospital, including 15 patients with keloid and 12 patients with hypertrophic Scars. Normal skin and peripheral blood were obtained from the keloid patients as self-control. Meanwhile pathological tissues and normal skin and peripheral blood were obtained from patients with hypertrophic scars as normal control.INTERVENTION:PCR-single strand conformation polymorphism was used to find the structure of the exons 7-9 in Fas gene in15 patients with keloid.MAIN OUTCOME MEASURES:The structure of the exons 7-9 in Fas gene.RESULTS:Heterozygous loss of Fas gene exon-8 was observed in all the 15 keloid patients, and 20% of them displayed an increase in exon-9 allele band.CONCLUSION: The genetic structure of Fas gene showed no mutation in hypertrophic scars, normal skin and the peripheral blood,but mutations were detected in exons-8 ,and -9 of Fas gene in keloid. This was closely related with the disfunction of its encoded proteins.

Objective To study the ingenious method to suture laparoscopic puncture holes.Methods A curved needle and a hook-like needle were used in 650 patients during biliary tract surgery.Results The curved needle was used in 200 patients and the hook-like needle was used in 450 patients.All patients had good wound healing except for one patient who developed puncture hole seroma under the xiphoid.The patient was treated with drainage and improved rapidly.No other complications such as puncture hole hemorrhage,infection and puncture hole hernia occurred.Conclusion Suturing using a curved needle and a hook-like needle was easy and effective,and avoided complications.

Objective To make a comparison of curative effect of 1 and 2 ureters anastomosis in modified sigma operation. Methods Modified sigma operation was used after radical cystectomy in 14 cases of bladder transitional cell carcinoma and 1 case of adenocarcinoma of the bladder.We used a folded suture of 25 cm from the sigmoid colon to make the U-shaped urinary reservoirs to anastomose with the ureters in study group.Among study group,1 ureter was anastomosed with urinary reservoir in 5 cases.In the other 10 cases,1.5-2 cm of the terminal parts of 2 ureters were sutured tI am not sure what the authors mean by big diamogether and then anastomosed with the urinary reservoir.Results 15 cases obtained good voiding control and no obvious water-electrolyte disturbance occurred.All the patients were cured and discharged within 15 to 22 days.Thirteen cases were followed-up in 8 to 24 month time period.Anastonmtic stenosis and moderate-severe hydronephrosis occurred in 4 cases with 1 ureter anastomosis.No hydronephrosis occurred in the 9 cases with 2 ureters anastomosis.Conclusions Modified sigma operation has the advantages of less injury and fewer complications.The patients can obtain self-control urination by anal sphincter without urine bags or intermittent catheterization,so the life quality of the patients is improved.Modified sigma operation with 2 ureters anastomosis has big diameter and there was a lower incidence of serious complications such as anastomotic stenosis.

Cholecystolithiasis with choledocholithiasis (CCL) is a common disease.The removal of common bile stone is a challenge for the surgery.This paper discussed the clinical application of three stone removal techniques including direct stone removal,irrigation and stone extraction by basket under cholangioscopy in order to take the stones effectively and safely,shorten the procedure time,avoid the injuries of common bile duct wall caused by the repetition of a single method such as biliary endoscopic stone extraction,reduce the difficulty of taking stone and enhance recovery of patients.

Objective To investigate the cytotoxicity of silica nanoparticles on vascular endothelial cells, and to clarify its action mechanism.Methods The 60 nm silica nanoparticle was selected and the invitro cultured human umbilical vein endothelial cells (HUVECs)were used as cell model.The HUVECs were divided into control and silica nanoparticle exposure groups with concentrations of 12.5,25.0,and 100.00 mg·L-1 .MTT assay was used for the determination of cell viability,lactate dehydrogenase (LDH)release assay for membrane integrity,flow cytometry (FCM)for intracellular reactive oxygen species (ROS)content,and real-time PCR assay for intracellular NF-E2-related factor 2 (Nrf2 ), heme oxygenase-1 (HO-1 ), superoxide dismutase 2 (SOD2 ) and glutamate-cysteine ligase catalytic subunit (GCLC)mRNA levels.Results The MTT results showed that the cell viabilities in each silica nnaoparticle exposure group were decreased compared with control group in a dose-dependent manner. Upon the silica nanoparticle exposure for 12 h,the cell viability was declined significantly only in 100 mg·L-1 exposure group compared with control group (P<0.05).When exposured for 24 h,the cell viabilities in 25.0, 50.0,and 100.0 mg·L-1 exposure groups were declined significantly compared with control group (P<0.05). Under the exposure to silica nanoparticle with the same dose, the cell viabilities were decreased along with the elongation of exposure time.LDH assay and FCM showed that except for that in 12.5 mg·L-1 exposure group, both the LDH activities in media and intracellular ROS levels in other exposure groups were increased compared with control group (P<0.05 ). The results of real-time fluorescence PCR showed that the mRNA levels of Nrf2, HO-1,SOD2 and GCLC in 100 mg·L-1 silica nanoparticle exposure group were increased significantly compared with control group (P<0.05).Conclusion Silica nanoparticles have toxicity to vascular endothelial cells,which includes reducing cell viability,membrane integrity destruction,induction of ROS generation,and tranSCriptional regulation of redox-related factors. Oxidative damage is one of the mechanisms of vascular endothelial toxicity mediated by silica nanoparticles.

Objective To study the effects of abiotic elicitors methyl jasmonate (MeJA) and salicylic acid (SA) on the alkaloids accumulation and related enzymes metabolism inPinellia ternata suspension cell cultures. Methods Using the leaf petioles-derived suspension cell cultures as the study object, the culture duration, concentrations of MeJA and SA were determined to get the optimal alkaloids accumulation, and the activities of metabolic enzymes IMP dehydrogenase and sAMP synthase were also measured.Results A 9-fold of dried biomass and a 3-fold of alkaloids accumulation were observed inP. ternata suspension cell cultures after culture for 21 d. Both MeJA and SA could significantly promote the accumulation of alkaloids inP. ternata suspension cells. 150 μmol/L MeJA enhanced alkaloids content (4.7 mg/gDW) by 3.6 folds in comparison with control group, whereas 50 μmol/L SA showed a 2.5-fold increase. Meanwhile, 100 μmol/L MeJA and 50 μmol/L SA promoted the increase in IMP dehydrogenase activity by 3.0 and 3.7 fold respectively, and 150 μmol/L MeJA and 100 μmol/L SA showed the increase by 2.6 and 4.4 fold respectively.Conclusion Proper adding exogenous MeJA and SA can promote the accumulation of alkaloids inPinellia ternata suspension cell cultures.

OBJECTIVETo investigate whether there are abnormal fibroblasts derived from the surrounding skin of keloids so that a more accurate therapy for keloids could be obtained.

METHODSSamples were taken for cell culture. When primary cells fully covered the culture bottle, the shape and distributing characteristics of fibroblasts were observed under the light microscope. 6-8 passage fibroblasts were selected for comparing the proliferating activity by MTT contrasting color method.

RESULTSThe fibroblasts have the same shape in all groups. But the fibroblasts derived from the surrounding skin grow crossly and overlapped just as the fibroblasts from keloids. The proliferating activity of the fibroblasts from surrounding skin is not as high as that from the border of keloids, but is higher than the normal skin fibroblasts derived either from a normal person or a patient with keloid.

CONCLUSIONIt is likely that there are abnormal fibroblasts in the surrounding skin of keloids.

Adolescent ; Adult ; Cell Division ; Cells, Cultured ; Female ; Fibroblasts ; pathology ; Humans ; Keloid ; pathology ; Male ; Skin ; pathology

ObjectiveTo identify the functions of the AP2/ERF family members in Pinellia ternata and promote the genetic improvement of P. ternata varieties. MethodWe identified and conducted a systematic bioinformatics analysis of the AP2/ERF family member genes in P. ternata based on the three generations of transcriptome data. Real-time polymerase Chain reaction　（Real-time） PCR was employed to determine the expression pattern of AP2/ERF genes in different tissues and under different stress conditions. ResultA total of eight full-length AP2/ERF family members were identified from the transcriptome data， which were classified into three sub-gene families： AP2， ERF， and DREB. The deduced AP2/ERF proteins in P. ternata had the length of 251-512 aa， the theoretical pI of 5.29-11.72， the instability index of 45.90-82.41， subcellular localization in the nucleus， and conserved domains and motifs. AP2/ERF genes were expressed in different tissues of P. ternata， with high expression levels in the leaf. The stress response experiments showed that PtERF1 mainly responded to NaCl stress. The expression of PtERF2 and PtERF4 was significantly up-regulated under low temperature and polyethylene glycol （PEG）-simulated stress. PtERF3 responded to both low temperature and NaCl stress. The expression of PtERF5 was induced by high temperature， low temperature， NaCl and PEG stress. The expression of PtERF7 was up-regulated under high temperature， while that of PtERF8 under low temperature. ConclusionThe AP2/ERF genes in P. ternata can respond to stress and have the potential functions of regulating photosynthesis and improving root stress resistance.