BACKGROUND:Wtih hyaluronic acid and col agen by cellautocrine as endogenous scaffolds, cellsheet technology can eliminate the disadvantages of exogenous scaffolds. Human adipose-derived stem cells combined with cellsheet technology used in the study of ful-thickness skin defects have become the current hot spot.
OBJECTIVE:To summarize the research progress in adipose-derived stem cellsheet.
METHODS:Articles related to adipose-derived stem cells, cellsheet technology and tissue-engineered skin were retrieved in VIP information and PubMed database from January 1993 to April 2014. The key words were“adipose-derived stem cell, wound healing, tissue-engineered skin, cellsheet”in English and Chinese, respectively. Total y 27 relevant articles were analyzed.
RESULTS AND CONCLUSION:Adipose-derived stem cells derived from discarded human adipose tissue are immmunocompatible and multi-potent, rendering them ideal for regenerative medicine applications. cellsheet technology seemingly provides several advantages over traditional tissue reconstruction with biodegradeable scaffolds. Adipose-derived stem cells combined with cellsheet technology can enhance tissue vascularization, promote wound healing, the secreted extracellular matrix provides the microenvironment for the celldifferentiation and growth, so it is expected to build a real three-dimensional tissue-engineered skin.

Objective To assess short-time effect of selective head cooling on neonatal hypoxic-ischemic encephalopathy induced by perinatal asphyxia. Methods Twenty-three infants of moderate/severe hypoxic ischemic encephalopathy (HIE) were divided into treatment group ( 14 cases) and control group (9 cases). The head hypothermia in the treatment group [rectal temperature ( 34. 0 ～ 35.0) ℃, nasopharyngeal temperature (34. 0 ± 0. 5 ) ℃]was induced by circulating water cooling cap for up to 72 h. Control group were treated routinely. All newborns were monitored and analyzed for blood pressure, heart rate, neurological function. Neonatal behavioral neurological assessment (NBNA) was conducted for assessment of neurobehavioral development on day 28 after birth. Results Treatment group showed slower hear rate as nasopharyngeal temperature decreased and there was significant difference as compared with the control group ( P ＜ 0.05 ). Apnea and arrhythmia were not found in either group. As compared with the control group, treatment group showed better results in seizure incidence, recovery of neonatal reflex( embracing reflection, sucking reflex and grasp reflex), and improvement of limb muscle tension. The NBNA score on day 28 was much higher in hypothermia group(35.00 ± 1.41 ) than that of control group(30. 67 ± 1.58) and there was significant difference (P ＜ 0.05). Conclusion Selective head cooling is safe for newborns with moderate/severe HIE if the nasopharyngeal temperature maintains at (34 :± 0. 5 ) ℃, rectal temperature at 34 ～ 35 ℃. Selective head cooling can effectively improve the short-term nervous symptoms and the neurobehavioral score,but the long-term efficacy remains to be further studied.

Objective To explore the procedures of breast repair and reconstruction after mastectomy with three methods for optimal appearances of the breast.Methods Three methods were adopted in this study:(1)Two-stage breast reconstruction operation with ampliate latissimus dorsi myocutaneous flap after mastectomy;(2)One stage breast reconstruction operation with transverse rectus adbominis myocutaneous(TRAM)flap after mastectomy;and(3)Two-stage breast reconstruction and nipple repair operation with tissue expander/implant after mastectomy preserving pectoralis major museulus or implants along under pectoralis major musculus.Results All of the flaps survived well and were well-shaped in total 10 patients(10 breasts)with a flap breast reconstruetion,in which the superior ratios were 83.33%.The appearance and volume were not satisfactory in 1 patiant treated with the method of tissue expander/implant,because the volume of tissue inflation was smaller than another one and 100 ml implant was applied.The partial flap necrosis occurred in 1 case with the method of TRAM flap.Conclusions With the development of plastic surgery technique,the appearances and shapes of reconstructed breast achieve the satisfactory effects.After the ampliate latissimus dorsi myocutaneous flap is applied,the appearance and volume of reconstructed breast are similar to the healthy one.The two-stage nipple repair operation is applied with breast reconstruction operation by the tissue expander/implant after mastectomy preserving pectoralis major musculus.According to the different methods of mastectomy to choose suitable procedures of breast reconstruction,the satisfactory appearances and psychotherapy will be achieved.

Our objective is to prevent doctors and nurses who worked on SARS ward from contracting this highly communicable disease through observing strict isolation strategies. The following measures have been taken in our hospitals. The wards for SARS patients were remodeled according to the isolation requirements for infectious diseases. They were separated into three quarters, namely clean area, semi-contamination area and contamination area. Two buffer zones were set up to separate the three quarters. Four specific passages were separately allotted to health care staffs, patients, soiled materials disposal and handling and dispatch of specimens, to avoid cross infection by contact. According to the criteria for isolation of serial A respiratory infectious diseases, every health care worker was strictly requested to wear and take off isolation garments properly when they entered and left the ward. Most importantly, specific garbs for different areas were essential for efficient isolation. Respiratory tract, skin and mucous surfaces should be effectively protected on the ward. In addition, doctors and nurses should wear special masks or helmets and waterproof suit during the performance of intubation, CPR or operation. Direction of air flow in SARS ward was diverted away from the clean area by using ventilator in order to prevent air-borne contamination. The air outlet of SARS ward was efficiently filtered to cut down pollution of the environment. Living quarters of medical staff working on SARS ward were isolated with a safe distance from the ward. Their health condition was closely monitored daily. After working in SARS ward for four weeks, they should enjoy a special vacation to assure their good health condition. In the past spring, 882 SARS patients were admitted to both the 309 hospital and Xiao Tanghan hospital. 1 632 health care workers worked for those SARS paitents by turns. To our satisfaction, none of our staffs was infected by SARS. Our experiences indicated that health care workers on the SARS ward could be effectively protected from contracting SARS infection by taking efficient and strictly observed protective measures.

Objective To study the treatment method of giant neurofibromatosis on body surface.Methods Replantation of the tumor skin,full-thickenss skin grafting and transferring of expanded flap were used to reconstruct the appearance and function after the excision of giant neurofibroma.Results Sixteen cases gained primary healing while other 2 cases with delayed healing after onestage or fractionated resection of giant neurofibroma on body surface.All the 18 patients resumed selfcare and working ability.Conclusions Giant neurofibromatosis has significant damage to the appearance and body shape and also greatly limits the patient's life and work.Patients can resume physical and psychological rehabilitation by surgical resection and plastic surgery treatment.

Objective To study the construction of tissue engineering adipose in vitro with silk fibroin scaffold and human umbilical cord mesenchymal stem cells (hUCMSCs) transfected by recombinant human insulin gene lentivirus.Methods hUCMSCs infected with recombinant lentiviral pLenti6.3-insulin-IRES-EGFP (Group A) by the best MOI=10 were seeded in silk fibroin scaffold; hUCMSCs transfected by EGFP gene (Group B)was regarded as negative control; and then the cell-scaf-fold complexes were cultured in adipogenic differentiation medium. MTT test was used to detect whether recombinant lentiviral affected the growth and proliferation of hUCMSCs and the growth activity of hUCMSCs seeded in silk fibroin.Results After 5-7 days for adipogenic culture,the numbers of fat-like cells in group A were significantly more than those in group B (P =0.007,P＜0.01).RTPCR results showed that the expression of PPARγ-2 in group A was much stronger than that in group B.MTT test showed that there was no significant difference in optical density (A)at each time between transfected group and nontransfected group (P=0.056,P＞0.05).And there was also no significant difference in optical density (A)between cell group and cell-scaffold group (P =0.066,P＞0.05).Conclusions Insulin gene could obviously promote hUCMSCs getting into adipose,and carrying recombinant human insulin gene lentiviral vector transfection of hUCMSCs and silk fibroin scaffolds could effectively construct tissue engineering adipose in vitro.

Objective To investigate the morbidity,clinical feature,diagnosis and therapy of ovarian disease in children.Methods Eighty-two children with ovarian disease were admitted and treated in Nanjing children's hospital from Jan.1992 to Jan.2007,were analyzed retrospectively with age,emergency admissions or not,dwell,pathology and method of operation.Results The age of 82 patients ranged from 1 day to 14 years old and the mean age was 6.7 years old.Thirty-one cases(37.8%) were emergency admissions and 51 cases(62.2%) were routine admissions.Twenty-seven cases(32.9%) were rural patients and 55 cases(67.1%) were urban patients.Forty-five cases(54.8%) were nontumorous disorder,31 cases(37.8%) were benign tumor and only 6 cases(7.4%) were malignant tumor.About the morbidity,12 patients(14.6%)were admitted from 1992 to 1996,24 patients(29.5%) from 1997 to 2001 and 46 patients(55.9%) from 2002 to 2007.Chemotherapy were carried out in 6 cases with malignant tumor in internal medicine,2 cases with sexual precosity kept observation,the others were cured.Conclusions Ovarian disease can occur at any age in children.The clinical manifestation is characterized mainly by acute abdomen.The incidence of ovarian disease of children in urban areas is higher than that children in rural areas.The morbidity continues to show an upward tendency and the pathologic manifestations are mostly benign,laparoscopic operation has obviously superiority.

OBJECTIVETo explore the clinical efficacy and mechanism of acupuncture and tapping therapy in the treatment of type 2 diabetes of yin deficiency pattern combined with stasis in the patients.

METHODSOne hundred and twelve patients were treated with acupuncture, followed by tapping method. The reinforcing method with lifting, thrusting and rotating needle technique was used at Taixi (KI 3), Shenmen (HT 7), Taibai (SP 3), Taiyuan (LU 9), Sanyinjiao (SP 6) and Zusanli (ST 36); and the even-needling technique was used at Taichong (LR 3), Hegu (LI 4), Xuehai (SP 10) and Fenglong (ST 40). The needles were remained for 30 min. After needle removal, the tapping method with plum-blossom needle was applied to Geshu (BL 17), Yishu (EX-B 3), Ganshu (BL 18), Shenshu (BL 23) and Pishu (BL 20), once every two days. The treatment of 3 months was required. The changes were observed before and after treatment in the relevant indices of fat-islet axis such as fasting plasma glucose (FPG), fasting leptin (FLP), fasting insulin (FINS), insulin sensitivity index (ISI), insulin resistance (Homa IR), insulin secretion index (Homa-beta) and the indices of obesity and blood lipids in the patients.

RESULTSAfter treatment, the total effective rate was 84.8% (95/112), in which the efficacy on the stasis predominated syndrome was better than that of yin deficiency predominated syndrome [92.9% (52/56) vs 76.8% (43/56), P < 0.05]. Before treatment, FPG, FLP, FINS, Homa-IR, total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), body mass, body mass index (BMI), fat percentage (F%) and obesity degree in the patients were higher obviously than those in the normal group (all P < 0.01); high-density lipoprotein cholesterol (HDL-C) and ISI were lower than those in the normal group (both P < 0.01). After treatment, FPG, FLP, FINS, Homa-IR, TC, TG, LDL-C, body mass, BMI, F% and obesity degree were all reduced obviously as compared with those before treatment (all P < 0.01). ISI and HDL-C were increased apparently (both P < 0.01). The efficacy on mild obesity was superior to that on moderate and severe obesity, and the efficacy in the young patients was better than that in the elderly, the efficacy in the males was better than that in the females, but the differences were not significant (all P > 0.05). The efficacy in the cases with 10 years duration of sickness was superior to those with over 10 years duration of sickness (P < 0. 01).

CONCLUSIONAcupuncture and tapping therapy achieves the significant efficacy on type 2 diabetes of yin deficiency pattern combined with stasis and its effect mechanism is related to the positive regulation of glucose, lipid metabolism and fat-islet axis.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Blood Glucose ; metabolism ; Cholesterol ; metabolism ; Diabetes Mellitus, Type 2 ; metabolism ; therapy ; Female ; Humans ; Insulin ; metabolism ; Leptin ; metabolism ; Male ; Middle Aged ; Treatment Outcome ; Triglycerides ; metabolism ; Yin Deficiency ; metabolism ; therapy

Objective:To explore whether cigarette smoking extract (CSE) has influence on fluorescence protein ex-pression of thrombomodulin (TM) on surface of African green monkey kidney fibroblast cells (COS-7) or not . Methods:When TM-green fluorescent protein (GFP) plasmid was successfully constructed ,COS-7 cells were trans-fected by it ,then incubated by prepared 5% CSE (5% CSE group) ,PBS of certain volume was added to serum-free minimal essential medium (MEM ,simple medium) ,which was cultured at the same time and treated as control group .Flow cytometry counting method was used to detect change of TM-GFP expression amount on COS-7 surface at different time point .Results:Compare with control group ,there were no significant difference in the expression of TM-GFP on COS-7 surface at 1h and 6h in 5% CSE group [1h :(134.99 ± 18.41) vs .(146.61 ± 12.06) ,6h :(116.89 ± 27.28) vs .(123.89 ± 39.24) ,P>0.05 both] .Conclusion:The 5% cigarette smoking extract has no influ-ence on fluorescence expression of thrombomodulin on surface of African green monkey kidney fibroblast cells .

Objective To investigate the effect of the serum of patients with hepatopulmonary syndrome on the myogenic differentiation,proliferation and migration of human lung fibroblasts.Methods The human lung fibroblasts were seeded in plates or flasks and randomly divided into 2 groups (n =31each) using a random number table:serum of patients with hepato-pulmonary syndrome group and serum of healthy volunteer group.The human lung fibroblasts were incubated in the DMEM culture medium containing 10％ serum of patients with hepatopulmonary syndrome or in the DMEM culture medium containing 10％ serum of healthy volunteers.At 24,48 and 72 h of incubation (T1-T3),the expression of smooth muscle-α-actin (SM-α-actin) and smooth muscle myosin heavy chain (SM-MHC) in human lung fibroblasts was determined by Western blot,the proliferation of the human lung fibroblasts was determined using 3H-TDR incorporation assay,and the migration of the human lung fibroblasts was determined by Transwell chamber assay.Results Compared with serum of healthy volunteer group,the expression of SM-α-actin and SM-MHC in human lung fibroblasts was significantly up-regulated at each time point,and the proliferation and migration of the cells were significantly enhanced at T2,3 in serum of patients with hepatopulmonary syndrome group (P＜0.05).Compared with the value at T1,the expression of SM-α-actin and SM-MHC in human lung fibroblasts was significantly up-regulated,and the proliferation and migration of the cells were significantly enhanced at T2,3in serum of patients with hepatopulmonary syndrome group (P＜0.05).Compared with the value at T2,the expression of SM-α-actin and SM-MHC in human lung fibroblasts was significantly up-regulated,and the proliferation and migration of the cells were significantly enhanced at T3 in serum of patients with hepatopulmonary syndrome group (P＜0.05).Conclusion The serum of patients with hepatopulmonary syndrome can promote the myogenic differentiation,proliferation and migration of human lung fibroblasts.