Objective To build artificial dermis by using the acellular dermis,collagen membrane and collagen gel as scaffolds.Methods The fibroblasts were isolated from infant skin.The 3~(rd) generation cells were seeded into 3 different scaffolds.The artificial dermis was detected by HE staining,phase contrast microscope or scanning electron microscope.Results The fibroblasts implanted on the ADM began to rupture and died after 2 to 3 days.Though the fibroblasts proliferated well in collagen gel,the artificial dermis contracted obviously.Another artificial dermis contracted slightly by inoculating fibroblasts on collagen membrane,and the fibroblasts on them were in appropriate proliferation.Conclusion The artificial dermis built by collagen membrane as scaffolds has a preferable structure for an ideal substitute of skin.

BACKGROUND: Ischemia/reperfusion can induce degenerative alterations in articular cartilage. However, the precise mechanism remains poorly understood. OBJECTIVE: To observe the morphological changes and the apoptosis of articular cartilage of femoral head epiphyses with ischemia/reperfusion. METHODS: A total of 80 Sprague-Dawley rats were randomly assigned to two groups: ischemia/reperfusion (model of ischemia/reperfusion in hip joint) and sham-surgery (exposure of abdominal aorta for 5 minutes) groups, with 40 animals in each group. Articular cartilages of femoral head epiphysis were col ected in 6, 12, 24, and 48 hours, 5 days, and 2 and 4 weeks after operation. Morphology of articular cartilage of femoral head epiphyses was examined by light microscope, and cel apoptosis was detected by TUNEL method. RESULTS AND CONCLUSION: Light microscopy showed chondrocytes degeneration and reduction, as wel as fibrosis in matrix of cartilage in the ischemia/reperfusion group. Chondrocyte apoptosis was observed in both groups by TUNEL. Several apoptotic cells, less than five, were observed in the sham-surgery, while 10-30 apoptotic cells were found in ischemia/reperfusion group at 48 hours. Results indicated that ischemia/reperfusion can induce degenerative changes in articular cartilage of femoral head epiphyses, and cel apoptosis in developing hip joint may participate in damage of articular cartilage. Inhibition of chondrocyte apoptosis in articular cartilage may be useful for the prevention and cure of early osteoarthritis.

Objective To observe the clinical effect and comfortable degree of the mode of super hair removal. Methods The mode of super hair removal was used to depilate the hair nearby the hair line, cheeks, upper lip, beard, ventrum, areola of breast, axillary cavity, extremities, bikini area and so on. The total number of sites was 1 000. Some sites that were especially susceptible to pain, for example, upper lip and buccal region, were smeared with compound lidocaine cream for 1 hour at least before treatment. Results Hairs in the areas of extremities, ventrum, back and axillary cavity generally needed 4 to 5 times to eradicate, and the patients had no evident discomfortableness; hairs near to the upper lip and lower mandible generally needed 5 to 7 times to reach the effect which the patient was content, and anesthetics was indispensable, or the patients would present discomfortableness. Conclusions The mode of super hair removal is more effective, quicker and more comfortable in comparison with conventional methods. Therefore, it deserves to be spread.

Objective To study the expression of long non-coding RNA (lncRNA)-urothelial carcinoma associated 1 (UCA1) and miR-34b in bladder cancer and its correlation to the clinicopathologic features of bladder cancer.Methods Between January 2011 and October 2012,the expression of UCA1 and miR-34b in 5 bladder cancer cell lines (T24,BIU-87,EJ,T24-MMC,T24-ADM) and 1 normal bladder cell lines (SV-HUC-1) were measured by real-time reverse transcription-polymerase chain reaction (RTPCR).Meanwhile,the 56 bladder cancer specimens and paraneoplastic normal bladder tissues,which diagnosed by pathology were collected from bladder cancer patients undergoing radical resection of bladder.Among them,41 cases were male and 15 cases were female.The mean age was (68.4 ± 7.5)years old,range 52 to 78 years.43 cases were older than 65 years old,and 13 cases were less than 65 years old.The pathological classification included non muscle-invasive bladder cancer (NMIBC) 18 cases,muscle-invasive bladder cancer 38 cases;low grade papillary urothelial carcinoma 22 cases,high grade papillary urothelial carcinoma 34 cases;12 cases were primary lesion,the other 44 cases were diagnosed as tumor recurrence.Real-time RT-PCR was performed to analyze the expression of UCA1 and miR-34b.Results The relative expression levels of UCA1 in the normal bladder cell lines (SV-HUC-1) and 5 bladder cancer cell lines (T24,BIU-87,EJ,T24-MMC and T24-ADM) were (0.0675 ± 0.0133),(0.2934 ± 0.0531),(0.4246 ± 0.0650),(0.4206 ± 0.0826),(0.6472 ± 0.0875) and (0.7165 ± 0.1032),respectively (P ＜ 0.05).Moreover,the expression levels of UCA1 were up-regulated in 2 drug resistant bladder cancer cells lines T24-MMC (0.6472 ± 0.0875)and T24-ADM (0.7165 ± 0.1032),as compared with the T24 bladder cancer lines (0.2934 ± 0.0531),respectively (P ＜ 0.05).However,the expression levels of miR-34b in 5 bladder cancer cell lines [T24 (0.1600 ± 0.0455),BIU-87 (0.1720 ± 0.0658),EJ (0.1150 ± 0.0352),T24-MMC(0.0576 ± 0.0087),T24-ADM (0.0510 ± 0.0125)] were decreased (P ＜ 0.05),as compared with normal bladder cell lines SV-HUC-1 (0.6384 ± 0.1083).Moreover,the expression levels of miR-34b were down-regulated in 2 drug resistant bladder cancer cells lines T24-MMC (0.0576 ± 0.0087) and T24-ADM(0.0510 ± 0.0125),as compared with the T24 bladder cancer lines T24 (0.1600 ± 0.0455),respectively (P ＜ 0.05).The relative expression levels of UCA1 and miR-34b in bladder cancer tissues and paraneoplastic normal bladder tissues were (0.4225 ± 0.0714) vs.(0.0532 ± 0.0192) and (0.0340 ± 0.0134)vs.(0.5643 ±0.0616),respectively (P ＜0.05).Statistical correlation analysis showed that UCA1 to be significantly negative correlated with miR-34b in bladder cancer specimens(r =-0.54,P ＜ 0.05).The high level of UCA1 and low level of miR-34b were significantly correlated with tumor malignant grade,invasiveness and recurrence.The 3-year overall survival rate (OS) in UCA1 (+)/miR-34b(-) group (27.6％) were significantly worse compared with non UCA1 (+)/miR-34b (-) group (73.7％).Conclusion High expression of UCA1 and low expression of miR-34b were associated with the occurrence and development of bladder cancer.

Objective To express and characterize the virus-like particles( VLPs) of H5 subtype containing of hemagglutinin ( HA ) and matrix 1 ( M1 ) protein by using Baculovirus-insect cells .Methods Full length genes encoding HA protein from the A/Indonesia/05/2005(H5N1) strain and the M1 protein from the A/Anhui/01/2005 ( H5N1 ) strain were cloned into a baculovirus expression vector to construct pFBD-M1-HA.The expression of HA and M1 proteins were detected by Western blot and indirect immunoflu-orescence after the transfection of Spodoptra frugiperda (Sf9) insect cells with recombinant baculovirus.Pu-rified VLPs were analyzed by SDS-PAGE and visualized with transmission electron microscope.The biologi-cal activity of purified VLPs was detected by hemagglutination test.Results The HA and M1 proteins of H5 subtype expressed by baculovirus-insect cells could be self-assembled into the functional mature VLPs.The hemagglutination titer of VLPs was as high as 1024 HAU/50μl.Conclusion The H5 subtype VLPs as pre-pared in this study would pave a way for the development of a candidate recombinant A ( H5) vaccine.

Objective The aim of this research was to produce apparatus of urinary diversion using silk-fibroin loading rab-bit adipose stem cells,and assess the effect of urinary diversion in a rabbit model.Methods Adipose stem cells were obtained and cultured in vitro,and flow cytometry analysis was performed to determine the adipose stem cells.These cultured adipose stem cells were used to seed on the silk-fibroin scaffolds,and after being incubated in the conditioned medium for 7 days,the a-bove compounds were made into apparatus of urinary diversion.This apparatus of urinary diversion was implanted into 20 rab-bits with radical cystectomy to develop urinary diversion.Five rabbits from each experimental group were euthanized at the spe-cific time points(1,2,3,4 months postoperatively),and the implants were harvested for histological and immunohistochemical a-nalysis.In the control group,silk-fibroins with unseeded cells(only silk-protein scaffolds)were also made into apparatus of uri-nary diversion and then used as urinary diversion on another 5 rabbits with the same process.Results Rabbits adipose stem cells were isolated and cultured successfully,and determined by flow cytometry.The silk-fibroin scaffolds were synthesized suc-cessfully.All rabbits were alive in the experimental group until the time of sacrifice.Histological and immunohistochemical anal-ysis showed multilayer uroepithelium coverage in the luminal surface of apparatus of urinary diversion,and as time went on,epi-thelial layers increased continuously.In the control group,all animals were dead within 3 weeks,and urine leakage,severe in-flammatory reaction and tissue destruction were found by autopsy.Conclusion The present experiment has successfully used silk-fibroin loading rabbit adipose stem cells to construct apparatus of urinary diversion,and demonstrates the feasibility of this kind of apparatus for urinary diversion in a rabbit model,which provides some experimental basis for clinical applications.

Objective:To establish donor liver quality related risk factors for the loss of function of transplanted liver.Methods:The data of donors and recipients of liver transplantation at the Organ Donation and Transplantation Center of the First Affiliated Hospital of Sun Yat-sen University from Nov 2011 to Dec 2018 were analyzed retrospectively. Propensity score matching (PSM) was performed to evaluate and screen the data of donors and recipients, in order to balance the covariates.Results:Of the organ donation, there were 70 males and 20 females , aging (40.6±16.3) years. Of the liver transplantation recipients, there were 70 males and 20 females , aging (41.8±20.3) years. Liver dysfunction after transplantation was significantly correlated with the following variables: the donor's CPR time( t=0.429, P=0.000), 15-minute retention rate of indocyanine green ( χ2=67.151, P=0.000), liver function grading ( χ2=54.154, P=0.000), bullae fatty liver grading ( χ2=8.120, P=0.017), vesicular fatty liver grading ( χ2=16.000, P=0.001), ICU stay time ( χ2=14.900, P=0.001)and serum creatinine level ( χ2=44.685, P=0.000). The donor scoring system was established in our studying. For the 90 organ donation cases, the donated liver quality were classified into four levels,which were of good correspondence to the prognosis of the recipients. Conclusion:This donor scoring system and grading standards established by analyzing the high-risk factors of liver dysfunction after transplantation helps evaluate the quality of donor liver in China.

Objective:To verify the clinical efficacy and safety of Qufeng Tongxuan method in treating low- and moderate-volume intracerebral hemorrhage at acute stage.Methods:A prospective, multicenter, randomized, double-blind, placebo-controlled study was performed; patients with low- and moderate-volume intracerebral hemorrhage at the basal ganglia and/or thalamus accepted treatment in 16 hospitals from September 2019 to April 2022 were enrolled. These patients were divided into experimental and control groups with a block randomized method by SAS software. Patients in control group were given conventional western medicine treatment; those in experimental group accepted Qufeng Tongxuan method (sequential therapy of Shexiang Huayu Xingnao granules and Zhilong Huoxue Tongyu granules) besides conventional western medicine treatment. NIHSS was used to assess neurological function before treatment and on 7 th, 14 th, 30 th, and 90 th d of treatment. Prognoses of these patients were assessed by modified Rankin scale (mRS) before treatment and on 180 th d of treatment. Brain CT was performed before treatment and on 7 th and 14 th d of treatment to calculate the hematoma volume. Before treatment and on 14 th d of treatment, changes of coagulation function, liver and kidney functions of the 2 groups were compared. Adverse reactions during treatment in the 2 groups were recorded. Results:No significant differences in NIHSS scores were noted between the 2 groups before treatment, on 7 th, 14 th, and 30 th d of treatment ( P>0.05); NIHSS scores in experimental group on 90 th d of treatment were signficantly lower than those in control group ( P<0.05); NIHSS scores in experimental group decreased gradually before treatment and on 7 th, 14 th, 30 th and 90 th d of treatment, with statistical significances ( P<0.05). No significant differences in mRS scores were noted between the 2 groups before treatment ( P>0.05); mRS scores in experimental group on 180 th d of treatment were signfciantly lower than those in control group ( P<0.05). No significant difference in hematoma volume was noted between the 2 groups before treatment and on 7 th and 14 th d of treatment ( P>0.05); both groups had gradually decreased hematoma volumes before treatment and on 7 th and 14 th d of treatment, respectively, with significant differences ( P<0.05); the volume difference of hematoma between 14 th d of treatment and before treatment in experimental group (6.42[4.10, 11.73]) was significantly higher than that in control group (4.00[1.25, 10.58], P<0.05). No significant differences in liver and kidney function indexes or coagulation function indexes were noted between the 2 groups before treatment and on 14 th d of treatment ( P>0.05). Adverse reaction incidence was 9.52% ( n=12) in experimental group and 10.34% ( n=12) in control group, without statistical difference ( P>0.05). Conclusion:Under premise of conventional western medicine treatment, Qufeng Tongxuan method can promote hematoma absorption and improve neurological deficit symptoms in low- and moderate-volume intracerebral hemorrhage at acute stage, without obvious adverse reactions.