Comparison of the Promoter Activity of vip3A and cry1Ia using IeGFP as a Fluorescent Reporter / 中国生物化学与分子生物学报

In spite of no homology in sequences‚ Vip3A and Cry1Ia toxins of Bacillus thuringiensis (Bt) share common characteristics‚ such as translocation across cell membranes after synthesis at the early stage of sporulation. The aim of the present study was to compare the regulation patterns and activities of the promoters of vip3A (P

A Rapid Protocol for Screening Genetically Modified Soybeans Based on Positive Plasmid Molecule Containing Multiple Targets / 中国生物化学与分子生物学报

Recently we witness the rising number of genetically modified (GM) soybean (Glycine max) events approved for importing from abroad and developed domestically, so it is urgent to establish a rapid screening protocol that can cover more events with less detection targets and fit the national condition. Additionally, in order to control the detection workload, it is also necessary to construct a multi-targets plasmid (MTP) molecule that can be used as the positive material. In this study, the information of the transgenic elements in 29 GM soybean events was collected and the combinations and frequencies of these elements were analyzed, to establish a novel screening protocol. It includes eight detecting targets, CaMV 35S promoter (P-35S), NOS terminator (T-nos), herbicide tolerance gene pat, E9 terminator (T-E9), insecticidal gene cry1Ac, AHAS promoter (P-AHAS), pin Ⅱ terminator (T-pin Ⅱ), and the event-specific sequence of the transgenic event DP305423, and an endogenous reference gene of soybean Lectin. After validation, the 29 GM soybean events described above can be screened by detection of the nine targets. This is referred to as the “8+1” protocol for GM soybean screening. Then these targeted sequences described in the protocol were simultaneously inserted into a cloning vector to construct the corresponding MTP pDDSC-1910. Finally, we tested whether it could be a positive plasmid. As expected, PCR analysis using pDDSC-1910 as a template showed that specific amplicons were observed with high sensitivity. Therefore, the “8+1” screening protocol for GM soybean was established, and the positive plasmid molecule pDDSC-1910 containing corresponding targets was successfully constructed. These results would facilitate the efficient screening and detection of transgenic soybeans.

Treatment of open fracture of lower limb in high altitude area / 中国骨伤

OBJECTIVE@#To explore the treatment methods and experience of open fracture of lower limb in high altitude area.@*METHODS@#From January 2016 to January 2021, 62 patients with open fractures of lower limbs were treated by staged surgery with the concept of injury control orthopedics, emphasizing wound treatment and combining various fracture fixation methods. There were 51 males and 11 females, ranging in age from 14 to 59 years old, with a mean of (37.2±12.3) years old; and the course of disease ranged from 7 to 59 days, with a mean of (23.7±15.5) days. According to Gustilo Anderson classification, there were 14 cases of typeⅠ, 24 cases of typeⅡ, 14 cases of typeⅢA, 8 cases of typeⅢB and 2 cases of typeⅢC. The fracture repair and wound healing were observed, and the clinical efficacy was evaluated by Johner-Wruhs evaluation standard.@*RESULTS@#Fifty-five patients were followed up, and the duration ranged from 4 to 36 months, with a mean of (14.7±8.5) months, and 7 cases were lost to follow-up. According to Johner-Wruhs evaluation criteria, 33 cases got an excellent result, 16 good, 4 poor and 2 bad. The wound healing was poor in 2 cases, partial necrosis of Achilles tendon in 1 case, nonunion of fracture in 1 case and delayed healing of fracture in 2 cases.@*CONCLUSION@#It is an effective method to treat the open fracture of lower extremity in high altitude area to pay attention to the management of soft tissue injury, the management of wound moisturizing, staged operation of fracture and full protection of blood supply at the fracture end. Paying attention to the treatment of soft tissue injury and the management of wound moisturizing, staged operation of fracture and full protection of blood supply at the fracture end are effective methods for the treatment of open fracture of lower limbs in high altitude areas.

Etanercept combined with Tripterygium wilfordii polyglycoside for treatment of rheumatoid arthritis in the elderly: a clinical study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of etanercept plus Tripterygium wilfordii polyglycoside (TWP) in elderly patients with active rheumatoid arthritis (RA).</p><p><b>METHODS</b>Totally 46 elderly patients with active RA were randomly assigned to the treatment group (22 cases) and the control group (24 cases). All patients received subcutaneous injection of etanercept, 25 mg each time, twice per week. The dosage was reduced to once per week 3 months later. Patients in the treatment group took TWP Tablet (10 mg each time, three times per day), while those in the control group took methotrexate (MTX), 10 mg each time, once per week. The whole course lasted for 24 weeks. Patients' rest pain, tender joint number, swollen joint number, health assessment questionnaire (HAQ), patients' global assessment, physicians' global assessment, erythrocyte sediment rate (ESR), C reactive protein (CRP), rheumatic factor were assessed at week 0, 4, 8, 12, and 24. The curative effect was statistically evaluated by the United States Institute of Rheumatology ACR20, ACR50, and ACR70 improvement criteria. Meanwhile, any adverse event was recorded and evaluated.</p><p><b>RESULTS</b>Totally 41 completed the trial, and 5 dropped off (3 in the treatment group and 2 in the control group). Compared with the control group, there was no statistical difference in ACR20, ACR50, or ACR70 in the treatment group (P > 0.05). Compared with before treatment in the same group, there was some improvement in tender joint number, swollen joint number, visual analogue scale (VAS) for patients' global assessment, VAS for physicians' global assessment, ESR, CRP, and HAQ between the two groups, showing statistical difference (P < 0.05). Compared with the control group in the same phase, there was no statistical difference in the treatment group (P > 0.05). There was no statistical difference in the occurrence of adverse events between the two groups.</p><p><b>CONCLUSIONS</b>Etanercept plus TWP could achieve equivalent therapeutic effect to that of Etanercept plus MTX. The two regimens could improve clinical signs, symptoms, and QOL related to RA. They were well tolerated in the treatment of elderly patients with active RA.</p>

Pain control after excimer laser corneal surface ablation / 国际眼科杂志(Guoji Yanke Zazhi)

By reshaping the cornea without the creation of a stromal flap, excimer laser corneal surface ablation eliminates flap-related complications and avoids the risk of ectasia that may occur after laser assisted in situ keratomileusis ( LASIK ) . Post-operative pain is one of the most significant disadvantages of surface ablation and thus the management of pain and discomfort following surface ablation is of great importance. We summarize mechanism of corneal pain and current approaches to pain management after surface ablation.

Continuous curvilinear capsulorhexis involving zonular area in manual small incision cataract surgery / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To evaluate the efficacy and safety of large sized continuous curvilinear capsulorhexis ( CCC ) involving zonular area in manual small incision cataract surgery ( MSICS) . METHODS:Totally 1 443 cataract patients ( 1 965 eyes ) underwent MSICS, in which large CCC up to 7-8mm were performed . The related conditions such as success rate of performing CCC, capsule rupture, corneal edema and visual acuity after surgery were recorded and statistically analyzed. RESULTS:In total of 1 965 eyes, 1 942 eyes ( 98. 83%) had successful CCC, 15 eyes (0. 76%) had a radial tear which could not be saved when performing CCC, 8 eyes (0. 41%) had anterior capsule opened with a cystotome or a scissor due to capsule membrane calcification. No posterior capsule rupture occurred, and intraocular lens was transplanted in all cases. Transient corneal edema was noted in 36 eyes (1. 83%). One day after surgery, 1650 eyes (83. 97%) had visual acuity ≥0. 5, 1867 eyes (95. 01%) ≥0. 3. No unstable intraocular lens was noted in all cases. CONCLUSION:Large CCC technique for MSICS is safe and reliable, not decreasing stability capsular bag and intraocular lens. It is of high originality despite zonular area is involved so that it is worthy of application in basic medical institutions.

Relationship between gene p53 codon 72 polymorphism and pathological scar formation after caesarean section / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the relationship between gene p53 codon 72 polymorphism and pathological scar formation occurrence after caesarean section.</p><p><b>METHODS</b>The method of molecular beacon with real-time PCR was applied to detect gene polymorphism of p53 codon 72 in blood samples taken from 303 pregnant women (within a week after caesarea section). The clinical visits were taken 3 times for 12th to 18th months to ascertain clinical formation of pathological scar and its relationship to genotype of p53. The chi-square method was used to analyze the relationship of p53 gene polymorphism and abnormal scar formation occurrence by statistical software SPSS 13.0.</p><p><b>RESULTS</b>Total of 303 pregnant women were assayed. 30 patients were found with pathological scar by clinical visit in the total 303 pregnant women. The genotype frequencies of total three types (C/C, C/G and G/G) of p53 gene codon 72 in patients with pathological scar are significantly different from that of normal pregnant woman. The frequency of C/C genotype in patients are higher than that of normal pregnant women (P < 0.01). The frequency of C/C genotype in these patients with pathological scar is higher (46.7%, 14/30) than C/G (33.0%, 10/30, P < 0.01) or G/G (20%, 6/30) genotype (P < 0.01). The C allele frequency in the patients is 63.7%. It is also higher than G allele (36.7%, P < 0.01). The OR value is 2.30. Therefore the C allele of p53 gene codon 72 is a risk factor for pathological scar.</p><p><b>CONCLUSIONS</b>There was a certain relationship between p53 gene codon 72 C allele and pathological scar formation after caesarean section.</p>

Preliminary study on inhibition of the hair follicle development by siRNA targeting Wnt10b in the cultured rat embryonic skin / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate whether the suppression of Wnt10b by siRNA could prevent the development of hair follicle in the cultured rat embryonic skin.</p><p><b>METHODS</b>siRNA-Wnt10b was synthesized by chemosynthesis method. The dorsal skin of SD rat at embryos were cultured in DMEM in the presence of different percentage of interfering RNA targeting Wnt10b. Wnt10b/beta-catenin expression was analyzed by real-time PCR everyday and by Western blot on the third day. The cultured embryonic skin underwent paraffin embedding, section, HE staining on the third day,in which the number of de novo hair follicle was calculated and statistically analyzed.</p><p><b>RESULTS</b>Wnt10b gene in the cultured embryonic skin could be knocked down with the siRNA-based method. Beta-catenin mRNA was not greatly influenced by the downregulation of Wnt10b mRNA. The number of de novo hair follicle placode in cultured embryonic skin decreased, along with the downregulation of Wnt10b and beta-catenin proteins expression.</p><p><b>CONCLUSIONS</b>The downregulation of Wnt10b mRNA and protein by siRNA reduces the number of de novo hair follicle placode in the cultured rat embryonic skin. Wnt10b may control cytoplasm beta-catenin concentration at the protein level.</p>

Construction of scaffold with human extracellular matrix from adipose tissue / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of constructing scaffold for tissue engineering with human extracellular matrix from adipose tissue.</p><p><b>METHODS</b>Fresh human adipose tissue was obtained by liposuction in 7 women who undergone liposuction. One part of the fat was used to isolate the adipose-derived stem cells (ADSCs), the other part was used to extract human extracellular matrix powder. After removing blood and oil components, the tissue was homogenized, centrifuged, freeze-dried, and crushed to powder by instrument. The structure of human ECM powder was observed with electron microscopy. The ADSCs were seeded and attached to the human extracellular matrix powder before and after labeled with fluorescent DiI, respectively. The adhesion rate was detected. The adhesion and growth of ADSCs were observed with Fluorescence microscope. The adhesion rate before and after DiI labeling was analyzed statistically with two-sample test of SPSS 13.0.</p><p><b>RESULTS</b>The ADSCs and human extracellular matrix powder were obtained successfully from adipose tissue. The ADSCs could be differentiated into adipose cells, bone cells and chondrocytes. SEM images showed that the power had both rugged and smooth surface with a porous structure characteristics. ADSCs could adhere to the scaffold easily, and the adhesion rate was (88.81 +/- 4.81)% and (86.48 +/- 4.58)% before and after DiI labeling. There was no difference between two groups. DiI labeled ADSCs were adhered to extracellular matrix scaffold and could grow in good condition.</p><p><b>CONCLUSIONS</b>Human adipose tissue extracellular matrix powder was easy to obtain, with diversity in size and shape which provided excellent substrates for cell adhesion and growth. It could be an ideal adipose tissue engineering scaffold.</p>

Experimental study of the effect of adipose stromal vascular fraction cells with VEGF on the neovascularization of free fat transplantation / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of adipose stromal vascular fraction cells (SVFs) with VEGF on the neovascularization of free fat transplantation.</p><p><b>METHODS</b>SVFs were obtained from subcutaneous fat and labelled with DiI. 0.3 ml autologous fat tissue was mixed with 0.2 ml cells: 1) autologous SVFs with VEGF (Group A); 2) autologous SVFs (Group B); 3) complete DMEM (Group C) And then the mixture was injected randomly under the back skin of 12 nude mice. The transplanted fat tissue in three groups was harvested at 2 months after implantation. Wet weight and diameter of fat grafts was measured. After HE and CD31 staining,blood vessel density, viable adipocytes and fibrous proliferation were observed.</p><p><b>RESULTS</b>Trace of SVFs labeled by DiI in vivo could be detected by fluorescent microscope. The wet weight of fat grafts was (191.90 +/- 9.81) mg in group A, (177.01 +/- 10.50) mg in group B, and (92.05 +/- 8.30) mg in group C (P<0.01). The diameter of fat grafts was (0.49 +/- 0.24) cm in group A, (0.40 +/- 0.26) cm in group B, and (0.32 +/- 0.28) cm in group C (P<0.01). Histological analysis showed the blood vessel density was (14.58 +/- 2.06)/HPL in group A, (11.55 +/- 2.18)/HPL in group B, (7.87 +/- 1.55)/HPL in group C. Compared with group B and group C, group A had more adipose tissue with less fat necrosis and fibrosis and had significantly higher capillary density.</p><p><b>CONCLUSIONS</b>The autologous adipose stromal vascular fraction cells with VEGF could improve the neovascularization of free fat significantly. It indicates a wide clinical application in the future.</p>

Experimental study on the best concentration of SVFs for promoting survival rate of fat graft / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of different concentrations of human adipose stromal vascular fraction cells (SVFs) on the survival rate of fat transplantation.</p><p><b>METHODS</b>0.3 ml fat tissue, derived and refined from clinical liposuction patients, was mixed with different concentrations of SVFs as 5 x 10(5)/ml in Group A, or 1 x 10(6)/ml in Group B, or 2 x 10(6)/ml in Group C, or completely medium in control group D. Then the mixture was injected randomly under the back skin of 6 nude mice. The transplanted fat tissue in four groups was harvested at 3 months after implantation. Wet weight of fat grafts was measured for macroscopic aspects. After HE staining, blood vessel density, viable adipocytes and fibrous proliferation were counted respectively for histological evaluation.</p><p><b>RESULTS</b>The wet weight of fat grafts in group B (81.670 +/- 7.528) mg was significantly higher than that in group A, C, D [(60.000 +/- 6.325) mg, (68.330 +/- 7.528) mg, (48.330 +/- 7.528) mg, respectively, P < 0.05)], but the difference between group A and group C was not statistically significant (P > 0.05). The grafts in group A, B and C had significantly higher blood vessel density than those in the control group D, whereas blood vessel density was the highest in group B (P < 0.05) and there was no significant difference between group A and C (P > 0.05). Compared with group A, C and D, histological analysis revealed that the fat grafts in group B was consisted predominantly of adipose tissue with less fat necrosis and fibrosis (P < 0.05). However, fibrosis counts were significant lower in group A, B and C than those in group D (P < 0.05), and there was no significant difference between group A and C (P > 0.05).</p><p><b>CONCLUSIONS</b>The human isolated SVFs has the advantages to improve the survival rate of fat transplantation, and the magnitude of 1 x 10(6)/ml is more practical and safe, indicating a wide clinical application in the future.</p>

Comparative adaptation of crowns of selective laser melting and wax-lost-casting method / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the marginal adaptation of crowns fabricated by selective laser melting (SLM) and wax-lost-casting method, so as to provide an experimental basis for clinic.</p><p><b>METHODS</b>Co-Cr alloy full crown were fabricated by SLM and wax-lost-casting for 24 samples in each group. All crowns were cemented with zinc phosphate cement and cut along longitudinal axis by line cutting machine. The gap between crown tissue surface and die was measured by 6-point measuring method with scanning electron microscope (SEM). The marginal adaptation of crowns fabricated by SLM and wax-lost-casting were compared statistically.</p><p><b>RESULTS</b>The gap between SLM crowns were (36.51 ± 2.94), (49.36 ± 3.31), (56.48 ± 3.35), (42.20 ± 3.60) µm, and wax-lost-casting crowns were (68.86 ± 5.41), (58.86 ± 6.10), (70.62 ± 5.79), (69.90 ± 6.00) µm. There were significant difference between two groups (P < 0.05).</p><p><b>CONCLUSIONS</b>Co-Cr alloy full crown fabricated by wax-lost-casting method and SLM method provide acceptable marginal adaptation in clinic, and the marginal adaptation of SLM is better than that of wax-lost-casting method.</p>

Establishment of molecular beacon real-time PCR for detecting p53 gene single nucleotide polymorphism / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a method based on molecular beacon real-time PCR for detecting single nucleotide polymorphisms (SNP) in codon 72 of scar-related p53 gene.</p><p><b>METHODS</b>Two fluorescence-labeled molecular beacon probes were synthesized targeting CCC/CGC SNP of p53 codon 72. The genomic DNA was extracted from the peripheral blood of 28 patients with keloid, and the CCC/CGC SNP of P53 gene codon 72 were assayed with molecular beacon real-time PCR. The results of SNP typing were compared with the results of reverse dot hybridization and confirmed by direct DNA sequencing.</p><p><b>RESULTS</b>The goodness of fit of this method was 100% in comparison with direct DNA sequencing, higher than that of reverse dot hybridization.</p><p><b>CONCLUSION</b>Molecular beacon real-time PCR is suitable for rapid clinical detection of SNPs in p53 gene.</p>

The cellular plasticity of human adipocytes / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the dedifferentiation phenomenon of human mature adipocytes cultured in vitro and to discuss the possibility of using dedifferentiation adipocytes (DA) as seed cells.</p><p><b>METHODS</b>Mature adipocytes and ASCs were harvested from human fat aspirates. Mature adipocytes were cultured and induced to DA by ceiling adherent culture method. Cell morphology were observed during the whole process. Viabilities of DA and ASCs were compared by MTT chromatometry and cell growth curves were drawn based on it. Cell surface markers of DA and ASCs were detected by flow cytometry. The adipogenic, osteogenic and chondrogenic ability of DA and ASCs were assessed by oil red O staining, alizarin bordeaux staining and alcian blue staining, respectively.</p><p><b>RESULTS</b>Human mature adipocytes can dedifferentiate into fibroblast-shaped DA. MTT chromatometry assay demonstrated that DA and ASCs both had strong reproductive activity, with no significant difference between them. Flow cytometry assay demonstrated that both DA and ASCs expressed HLA-ABC, CD29 and CD44, while didn't express CD45, CD34 and CD106. After two weeks of adipogenic differentiation, lipid droplets could be displayed by oil red O staining in both DA and ASCs. After two weeks of osteogenic differentiation, calcium salts mineralization in DA and ASCs could be detected by alizarin bordeaux staining. After two weeks of chondrogenic differentiation, matrix of cartilage cells in DA and ASCs could be detected by alcian blue staining.</p><p><b>CONCLUSIONS</b>Mature adipocytes can be dedifferentiated into DA in vitro. DA has strong reproductive activity, as well as osteogenic, chondrogenic ability and strong adipogenic ability. It expresses some of the stem cell-related cell surface proteins and is a promising seed cell for adipose tissue engineering.</p>

Effect of liposome-mediated HSV-tk gene transfer on fibroblast apoptosis in rats with scald injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the effect of HSV-tk gene transfer on the apoptosis of fibroblast in rats with scald injury.</p><p><b>METHODS</b>The recombinant eukaryotic expression vector pcDNA3.1/tk was transfected via liposome in the skin of rats with scald injury. The expression of tk gene was detected by RT-PCR technique, and after GCV injection, the apoptosis of the fibroblasts positive for tk gene was observed under transmission electron microscope.</p><p><b>RESULTS</b>Liposome-mediated HSV-tk gene transfection of the rat skin resulted in the positive expression of tk gene in the fibroblasts in the burn wound. GCV injection induced the apoptosis of the positively transfected fibroblasts.</p><p><b>CONCLUSION</b>HSV-tk gene transfer mediated by liposome can promote the apoptosis of the fibroblasts in rats with scald injury.</p>

Study on cotransfection of genes of insulin-like growth factor I and herpes simplex virus thymidine kinase for optimization of wound healing / 中华烧伤杂志

<p><b>OBJECTIVE</b>To study the effect of cotransfection of genes of insulin-like growth factor I (IGF-I) and herpes simplex virus thymidine kinase (HSV-tk) on wound healing.</p><p><b>METHODS</b>Thirty male Wistar rats were inflicted with 30% TBSA full-thickness scald. They were then divided into A group (4.6 microg pcDNA3.1/IGF-I+Lipofectamine 2000+saline), B group (3.6 microg pcDNA3.1/HSV-tk+Lipofectamine 2000+saline), C1 group and C2 group (2.3 microg pcDNA3.1/IGF-I+1.8 microg pcDNA3.1/HSV-tk+Lipofectamine 2000+saline), and D group (3.0 microg pcDNA3.1+Lipofectamine 2000+saline) according to the random number table, with 6 rats in each group. The above-mentioned mixtures were subcutaneously injected into left back of each rat the moment after injury and on post scald day (PSD) 7, 14, 21, and 28. Gancyclovir (2.5 mg/100 g) was hypodermically injected into rats in C2 group on PSD 29, 30, 31, 32. Changes in body weight of rats were measured. Wound healing rates were calculated. On PSD 35, the expressions of IGF-I gene in local wound and liver tissue were determined with immunohistochemical staining. The serum expression of IGF-I was determined with radioimmunoassay. Expression of HSV-tk gene in local wound was determined with RT-PCR. Apoptosis of fibroblast in C1 and C2 groups was observed under transmission electron microscope. Data were processed with one-way analysis of variance and Turkey method.</p><p><b>RESULTS</b>Body weight of rats in A, C1, and C2 groups increased from PSD 7 through 35, and the difference between former three groups and B, D groups was statistically significant (with F value respectively 2.764, 4.519, 5.009, 13.449, 5.877, P values all below 0.05). Wound healing rates of rats in A, C1, and C2 groups were higher than those in B, D groups (with F value respectively 5.286, 100.880, 152.380, 127.850, 147.750, P values all below 0.05). IGF-I gene was positively expressed in wound fibroblast in A, C1 and C2 groups, but negatively in liver tissues of all the rats. There was no significant statistical difference among groups in serum content of IGF-I [from (1185+/-170) to (1270+/-130) ng/mL, F=0.355, P=0.838]. HSV-tk gene was positively expressed in rat skin tissue in B, C1 and C2 groups. Fibroblast apoptosis was observed under transmission electron microscope in C2 group, but it was not observed in C1 group.</p><p><b>CONCLUSIONS</b>Cotransfection of pcDNA3.1/IGF-I and pcDNA3.1/HSV-tk mediated by liposome can promote wound healing, and inhibit the scar proliferation to some extent.</p>

Transplantation of adipose tissue-derived stem cells promotes soft tissue wound repair in rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the effect of local and intravenous transplantation of adipose tissue-derived stem cells (ADSCs) in promoting soft tissue wound healing in rats.</p><p><b>METHODS</b>ADSCs isolated from the adipose tissues of SD rats were cultured in vitro, and the third-passage cells were identified for their capacity of multipotent differentiation. Eighteen SD rats with 1.8 cm² dorsal full-thickness soft tissue defects (0.5 cm deep) were randomized into 3 groups to receive injection of 3.0×10⁶ DiI-labeled ADSCs via the tail vein, local injection of the cells at the wound site, or injection of saline (control). The wound healing was evaluated on days 3, 7, 11, and 14 postoperatively. On day 24 after the injury, tissue samples at the wound site were collected for fluorescent microscopy and HE staining.</p><p><b>RESULTS</b>The ADSCs obtained were capable of adipogenic, osteogenic, and neurogenic differentiation in vitro. ADSCs transplantation significantly promoted wound healing as compared to the control group. Obvious wound contracture was observed in the local injection group on day 3 and in the intravenous injection group on day 7. Fluorescence microscopy revealed DiI-positive cells in the healing wound, and HE staining showed a greater tissue thickness at the wound in the two ADSCs transplantation groups. Compared to the control group, the two ADSCs transplantation groups showed more gland-like structures and better neovascularization at the wound.</p><p><b>CONCLUSION</b>ADSCs can significantly promote wound healing in rats, and local injection of ADSCs allows more rapid and obvious wound healing than tail veil injection of the stem cells.</p>

The differences between adipose tissue derived stem cells and lipoma mesenchymal stem cells in characteristics / 中华整形外科杂志

<p><b>OBJECTIVE</b>To compared the biological characteristics of adipose-derived stem cells and lipoma-derived mesenchymal stem cells (LMSCs) in vitro, so as to assess the safety of adipose-derived stem cells( ASCs) for transplantation.</p><p><b>METHODS</b>Regular slice and stain of adipose and lipoma tissue were performed. ASCs and LMSCs were isolation from the two tissues by enzymatic digestion, and the appearance of the cultured cells was observed. The cell viability was evaluated with MTS chromatometry and cell growth curve was generated. Flow cytometry was performed for cell cycle analysis and the expression of the cell surface marker profiles. QRT-PCR was used to detect the expression of tumor-specific gene (the high-mobility group AT-hook 2, HMGA2), and immunocytochemistry was used to detect the expression of telomerase.</p><p><b>RESULTS</b>Marked difference was observed in histologic sections of adipose tissue and lipoma tissue. ASCs showed a good consistent in cell morphology while LMSCs not. ASCs showed a significant lower proliferation capacity than LMSCs by MTS chromatometry (P = 0.000). The expression of CD29, CD44, CD105 was similar in ASCs and LMSCs, while the level of CD133 was significantly lower in ASCs (5.35%) than in LMSCs (26.87%). The expression of HMGA2 was lower in ASCs (RQ = 1) than in LMSCs (RQ = 1.79) by qRT-PCR, it has statistically difference between them (P < 0.01); And in ASCs and LMSCs, the integrated optical intensity (IA) values of hTERT expression are 1379.597 +/- 498.617 and 3 328.108 +/- 902.856, size (area) are 132,390.27 +/- 35,568.945 and 238,000.53 +/- 49,264.289, density (mean) are 0.009 +/- 0.003 and 0.014 +/- 0.003, revealed the expression of hTERT also shown a significant lower level in ASCs than in LMSCs by immunocytochemistry.</p><p><b>CONCLUSIONS</b>It indicates significant difference between ASCs and LMSCs in the biological characteristics in vitro. There is no evidence of malignant transformation of ASCs.</p>

Experimental study of the effect of adipose stromal vascular fraction cells on the survival rate of fat transplantation / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of adipose stromal vascular fraction cells (SVFs) on the survival rate of fat transplantation.</p><p><b>METHODS</b>0. 5 ml autologous fat tissue was mixed with: 1) Di-labeled autologous SVFs ( Group A); 2) Di-labeled autologous adipose-derived stem cells (ASCs) (Group B); 3)Complete DMEM (Group C). And then the mixture was injected randomly under the back skin of 14 rabbits. The transplanted fat tissue in three groups was harvested at 6 months after implantation. Wet weight of fat grafts was measured for macroscopic aspects. After HE staining, blood vessel density, viable adipocytes and fibrous proliferation were counted respectively for histological evaluation. Trace of DiI-labeled ASCs in vivo was detected by fluorescent microscope.</p><p><b>RESULTS</b>The wet weight of fat grafts in group A (291.0 +/- 72.1) mg and group B (269.3 +/- 67.3) mg was significantly higher than that in group C (177.8 +/- 60.0) mg, but the difference between Group A and Group B was not significant. Histological analysis revealed that the fat grafts in group A and B was consisted predominantly of adipose tissue with less fat necrosis and fibrosis, compared with the fat grafts in group C. The grafts in both group A and B had significantly higher capillary density than those in the control group. Part of vascular endothelial cells were observed to origin from ectogenic DiI-labeled SVFs and ASCs.</p><p><b>CONCLUSIONS</b>The autologous isolated SVFs has a similar effect as autologous cultured ASCs to improve the survival rate of fat transplantation. And the former is more practical and safe, indicating a wide clinical application in the future.</p>

Preliminary comparison study of adipogenic differentiation capacity between dedifferentiated adipocytes cells and adipose-derived stem cells in vivo / 中华整形外科杂志

<p><b>OBJECTIVE</b>To compare the adipogenic differentiation capacity of dedifferentiated adipocytes cells (DA) and adipose-derived stem cells (ASCs) in vivo, so as to select good adipogenic seed cells for tissue engineering.</p><p><b>METHODS</b>Mature adipocytes and ASCs were isolated by means of enzymatic digestion from the liposuction aspirate. Then the DA cells were acquired by ceiling adherent culture of mature adipocytes and the 3rd passage cells were used. The DA cells and ASCs were cultured with fibrin glue in vitro respectively. The compatibility of scaffold with cells was detected by microscopy and scanning electron microscopy. The scaffold-cell composite was also labeled by DiI. The composite was injected subcutaneously on the nude mice back, respectively (DA-FG group, n = 8; ASCs-FG group, n = 8; sham FG group, n = 8). 8 weeks after implantation, the newly formed tissue was taken out for general observation and histologic study.</p><p><b>RESULTS</b>Mature adipocytes were transferred to DA cells with spindle shape, like fibroblast. The ASCs were also spindle. Three days after culture of cell-scaffold composite in vitro, the cells grew well. 8 weeks after implantation, the newly formed tissue was found under the skin both in DA-FG and ASCs-FG groups, but not in sham FG group. The newly formed tissue was mature fat tissue and originated from the seed cells. The average wet weight of the new-formed tissue was higher in DA-FG group than that in ASCs-FG group. The average fibrosis ratio was lower in DA-FG than in ASCs-FG group.</p><p><b>CONCLUSIONS</b>The tissue-engineered adipose tissue can be achieved with DA cells and ASCs as seed cells. Compared with ASCs, the new-formed fat tissue with DA has a higher wet weight and lower fibrosis ratio.</p>