Comparison between case management and group work on rehabilitation effects in discharged schizophrenia patients / 上海预防医学

Objective:To compare the intervention effect of Case Management and Group Work in discharged schizophrenic patients. Methods:A total of 100 patients with stable schizophrenia who were discharged from a mental health center in a district of Shanghai were randomly divided into two groups： case management （CM） and group work （GW）， with 50 patients each. The group work method and case management model were used， respectively， to provide psychiatric symptom assessment，medication guidance，functional training，health education and other services to for 6 months. Both Positive and Negative Syndrome Scale （PANSS） and Social Disability Screening Schedule （SDSS） were used for assessment at the beginning and at the end of 6th month. The conditions and the improvement of social function of these two groups were compared. Results:Before and after the intervention，the positive symptom score，negative symptom score and PANSS total score of the CM group were decreased，the difference was statistically significant（t=4.214，3.926，3.929，P<0.001）. The PANSS score of the GW group only had significant difference before and after the general pathology and the total score（t=2.195，2.466，P<0.05）.There were significant differences in SDSS scores between these two groups before and after the intervention（P＜0.001）. There was a significant difference in the reduction rate of positive symptom score between the two groups（z=-2.937，P<0.05）. There was also a significant difference in the reduction rate of SDSS scores（z=-3.834，P<0.001）. Conclusion:Both case management and group work can stabilize the condition of schizophrenia patients and improve social function，but there is a slight difference in the emphasis of the two methods.

Discovery of Eucalyptin C, derived from the fruits of Eucalyptus globulus Labill., as a novel selective PI3Kγ inhibitor for immunosuppressive treatment / 中国天然药物

The fruits of Eucalyptus globulus Labill. are known to have a plenty of medicinal properties, such as anti-tumor, anti-inflammatory, and immunosuppressive activity. Our previous study found that the phloroglucinol-sesquiterpene adducts in the fruits of E. globulus were immunosuppressive active constituents, especially Eucalyptin C (EuC). Phosphoinositide 3-kinases-γ (PI3Kγ) plays a pivotal role in T cell mediated excessive immune responses. In this study, EuC was first discovered to be a novel selective PI3Kγ inhibitor with an IC

DNA aptamer selection in vitro for determining ketamine by FluMag-SELEX / 法医学杂志

OBJECTIVE@#To select specific DNA aptamer for determining ketamine by FluMag-SELEX.@*METHODS@#Based on magnetic beads with tosyl surface modification as solid carrier and ketamine as target, a random ssDNA library with total length of 78 bp in vitro was compounded. After 13 rounds screening, DNA cloning and sequencing were done. Primary and secondary, structures were analyzed. The affinity, specificity and Kd values of selected aptamer were measured by monitoring the fluorescence intensity.@*RESULTS@#Two ssDNA aptamers (Apt#4 and Apt#8) were successfully selected with high and specific abilities to bind ketamine as target with Kd value of 0.59 and 0.66 μmol/L. The prediction of secondary structure was main stem-loop and G-tetramer. The stem was the basis of stability of aptamer's structure. And loop and G-tetramer was the key of specific binding of ketamine.@*CONCLUSION@#FluMag-SELEX can greatly improve the selection efficiency of the aptamer, obtain the ketamine-binding DNA aptamer, and develop a new method for rapid detection of ketamine.

Construction of the tissue engineering seed cell (HaCaT-EGF) and analysis of its biological characteristics

OBJECTIVE@#To construct the tissue engineering seed cell (HaCaT cell line) with stable expression of the human epidermal growth factor (EGF), and analyze the changes of its biological characteristics.@*METHODS@#PCDNA3.1-EGF eukaryotic expression vector was transferred into HaCaT cell, and G418 was utilized to select the HaCaT-EGF cell line. Using an inverted microscope, PCR, ELISA method to detect the changes of the cell morphology, the expression of the EGF gene and protein, and the mRNA expression levels of apoptosis related molecule Caspase-3, the cell cycle related protein cyclin D1.@*RESULTS@#The mRNA expression levels of the obtained HaCaT-EGF cell were more than 100 times higher than the level of ordinary HaCaT cell. The colony of the HaCaT-EGF cells was more focused and tight compared to the empty vector transfected HaCaT cells and normal HaCaT cells. The expression levels of apoptotic factor Caspase-3 and cyclin D1 in HaCaT-EGF cell were significantly higher than those in the empty vector HaCaT- pcDNA3.1 cell, and the differences were statistically significant (P0.05).@*CONCLUSIONS@#HaCaT-EGF cell can continuously secrete EGF, and the biological characteristic is stable. It can be used for tissue engineering experiment and is an ideal seed cell for constructing tissue engineered skin.

Systemic therapy for defects of skin and soft tissue on the knees after severe trauma or burn / 中华烧伤杂志

<p><b>OBJECTIVE</b>To explore the methods of systemic treatment of defects of skin and soft tissue on the knees after severe trauma or burn.</p><p><b>METHODS</b>Twenty patients with defects of skin and soft tissue on the knees after severe trauma or burn hospitalized in our center from January 2009 to December 2011. The injury areas on the knees ranged from 5 cm×4 cm to 30 cm×20 cm. The wounds were treated with radical debridement, vacuum sealing drainage, and douche through dripping to control infection in early stage. Then they were covered with transplantation of skin grafts plus flap or only with flap. Totally 8 local flaps (including 6 local rotation or transposition flaps and 2 saphenous artery flaps) and 12 free flaps (including 8 anterolateral thigh flaps and 4 latissimus dorsi musculocutaneous flaps) were used. The flap size ranged from 6 cm×5 cm to 32 cm×22 cm. The rehabilitation training of the knee joints was carried out in the early stage after wound healing.</p><p><b>RESULTS</b>All free skin grafts and flaps used in 15 patients survived. Thirteen of them were primarily healed, while some small parts of skin grafts of the other two patients were in poor condition because of infection, and they healed after another session of skin transplantation. Infection occurred under the free flap in one of the 5 patients transplanted with flaps only, which was healed after continuous douche through dripping and another surgical debridement following wet dressing. The knee joints were in good function during the follow-up period of 1 - 3 years.</p><p><b>CONCLUSIONS</b>The systemic therapy of radical debridement, vacuum sealing drainage technique, douche through dripping, transplantation of large autologous grafts and flaps, and the early rehabilitation training are effective and reliable in repairing defects of skin and soft tissue at the knee region after severe injuries.</p>

Effects of adipose-derived stem cells on renal injury in burn mice with sepsis / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the effect of adipose-derived stem cells (ADSC) on renal injury in mice with burn injury and sepsis and its underlying mechanism.</p><p><b>METHODS</b>(1) Adipose tissue was collected from both inguinal regions of 5 C57BL/6J mice to isolate, culture and purify ADSC through enzyme digestion, density gradient centrifugation, and adherence method. Cells of the third passage were used in the experiment. The morphologic change in cells was observed and the growth curve of cells was determined. The expression of cell surface antigen phenotype was analyzed by flow cytometry, and the cells were identified by adipogenic and osteogenic differentiation. (2) Another 37 C57BL/6J mice were divided into normal control group (n = 5), saline group (n = 16), and group ADSC (n = 16) according to the random number table. The mice in saline group and group ADSC were injected with Pseudomonas aeruginosa after being subjected to 15% TBSA full-thickness burn on the back to reproduce septic burn model. Then the mice were injected with saline and ADSC through tail vein respectively. At post burn hour (PBH) 12, 24, 48, and 72, the pathological change in kidney tissue was observed, the levels of blood urea nitrogen and serum creatinine were determined, and the levels of TNF-α, IL-12, IL-10, and cyclooxygenase-2 (COX2) mRNA were determined with real-time fluorescence quantitative PCR in both groups. Above-mentioned indexes were also examined in the normal control group (without burn). Data were processed with multifactor analysis of variance and LSD- t test.</p><p><b>RESULTS</b>(1) Cells in the third passage were orderly arranged with the shape similar to fibroblasts. The percentages of CD90(+), CD105(+), CD34(-), and CD45(-) cells were all above 90%. The cells could differentiate into osteoblasts and adipocytes. The cells were identified to be ADSC. (2) From PBH 12 to PBH 72, the neutrophil infiltration gradually increased, and the structure of kidney tubules and glomeruli were deranged in saline group. The pathological change in kidney tissue in group ADSC was less serious than that of normal control group at each time point. From PBH 12 to PBH 72, the levels of blood urea nitrogen and serum creatinine in saline group were significantly higher than those of normal control group and group ADSC (P values all below 0.01). Compared with those of the normal control group, the levels of TNF-α and IL-12 mRNA were higher in group ADSC and saline group at PBH 24 (P values all below 0.05). At PBH 24, the level of TNF-α mRNA in group ADSC (1.58 ± 0.19) was lower than that of saline group (3.36 ± 0.30, P < 0.05). At PBH 24, the levels of IL-10 and COX2 mRNA in group ADSC (2.89 ± 0.47, 4.90 ± 0.59) were higher than those in normal control group (1.00 ± 0.15, 1.00 ± 0.27) and saline group (1.32 ± 0.38, 1.57 ± 0.38, P values all below 0.05).</p><p><b>CONCLUSIONS</b>ADSC can decrease the levels of blood urea nitrogen and serum creatinine, promote the production of anti-inflammatory cytokines IL-10 and COX2, and reduce the release of the pro-inflammatory cytokines TNF-α and IL-12 to offer protective effects against renal injury in burn mice with sepsis.</p>

Reproduction of a mouse model of deep partial-thickness scald and determination of hypoxia in the wound / 中华烧伤杂志

<p><b>OBJECTIVE</b>To reproduce a stable mouse model of deep partial-thickness scald and to determine the hypoxia status in the wound.</p><p><b>METHODS</b>(1) A homemade scald-producing apparatus with constant steam (92 °C) emission was used to reproduce scald injury on the back (2 cm in diameter) in 80 male BALB/c mice for different duration (2, 4, 6, and 8 s), with 20 mice for each scald duration. The nozzle was aligned perpendicularly to the back of mice, 2 cm above the skin surface. The gross condition of wound was observed with naked eyes immediately after injury. Skin samples of 5 mice with different burn duration were harvested 0, 12, 24, and 48 h after scald for histopathological observation with hematoxylin and eosin staining, to screen the scalding time and time for biopsy of scalded skin to determine proper scalding time for the experiment. (2) Model of deep partial-thickness scald was reproduced with the desired scalding time as shown in the preliminary experiment in another 5 BALB/c mice. The hypoxia status in subcutaneous tissue was observed with immunohistochemical staining 72 h after scald. Another 20 BALB/c mice were divided into normal control group (n = 5, without scald) and deep partial-thickness scald group (n = 15, scalded for a suitable duration as determined in the preliminary experiment) according to the random number table. The subcutaneous oxygen content in wound center, the margin of the wound, and the normal skin adjacent to the wound was detected with laser Doppler transcutaneous oxygen tension 72 h after scald, with 5 mice in each region. Data were processed with one-way analysis of variance.</p><p><b>RESULTS</b>(1) The wound of mice with different scald durations was pale, clean, and no exudate was observed right after injury. (2) The burn depth developed gradually along with the scalding time and sample harvesting time, and it became stable 24 h after scalding. A deep partial-thickness injury was observed in the dermis of mice scalded for 4 s and harvested 24 h after scald, and it was shown that the external hair sheath was still present, and it was determined to be a deep partial-thickness scald. (3) Dense staining of pimonidazole (hypoxia) was found in deep partial-thickness scald wound 72 h after scald, especially in the marginal zones of the wounds. The partial oxygen pressure in the wound center, wound margin, and normal skin around the wound was respectively (36.2 ± 3.2), (37.0 ± 1.4), (37.4 ± 2.7) mm Hg (1 mm Hg = 0.133 kPa), showing no statistically significant difference among them (F = 74.705, P > 0.05), but they were significantly lower than that of the control group [(53.1 ± 2.4) mm Hg, with F values respectively 82.377, 91.375, 100.531, P values all below 0.05].</p><p><b>CONCLUSIONS</b>Deep partial-thickness scald model can be reproduced in (20.0 ± 1.0) g male BALB/c mice by scalding with 92 °C hot steam for 4 s, and the depth of wound becomes stable 24 h after scalding. Hypoxia can be found in the scalded wounds, especially in the marginal zones of the wounds.</p>

Influence of negative pressure wound therapy on the angiogenesis of wounds in diabetic rats / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the influence of negative pressure wound therapy on the angiogenesis of wounds in diabetic rats.</p><p><b>METHODS</b>Diabetes model was reproduced by intraperitoneal injection of 20 g/L streptozotocin in the dosage of 65 mg/kg in 40 SD rats. Two weeks later, rats were divided into control group (C) and negative pressure group (NP) according to the random number table, with 20 rats in each group. A piece of full-thickness skin in the center of the back of each rat in the size of 2 cm×2 cm was excised to produce a wound. Immediately after injury, wounds in group C were given conventional dressing change; wounds in group NP were treated with continuous negative pressure (-16.0 kPa) therapy for four hours a day, which lasted for seven days. (1) Blood glucose and body weight of rats in two groups were respectively measured by glucose meter and electronic scale before treatment, and 1 and 2 week (s) after. (2) Wound blood flow was detected by laser Doppler perfusion imager before treatment and on post treatment day (PTD) 1, 3, 7, with 5 rats at each time point. (3) On PTD 3 and 7, respectively, five rats from each group were sacrificed. The wound tissue was excised and divided into two parts. The angiogenesis in the left part tissue was observed with immunohistochemical staining. The microvessel density was calculated. (4) The full-thickness skin excised before treatment and the right part tissue freeze on PTD 3 and 7 were collected. On PTD 1 and 14, wound tissue was excised in the above-mentioned method. The mRNA levels of the vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 1 (Fit-1), angiopoietin 1 (Ang-1), Ang-2, and tyrosine kinase receptor 2 (Tie-2) were determined with real-time fluorescence quantification PCR. Data were processed with two-way analysis of variance or LSD-t test.</p><p><b>RESULTS</b>(1) No significant difference was observed between two groups in blood glucose level and body weight as a whole or at each time point (with F values respectively 0.667, 0.176, t values from 0.311 to 0.707, P values all above 0.05). (2) The difference in the overall wound blood flow of rats between two groups was significant (F = 24.66, P < 0.05). On PTD 1, 3, 7, values of wound blood flow of rats in group NP were (179 ± 24), (219 ± 12), (192 ± 30) perfusion unit, significantly higher than those of rats in group C[(127 ± 16), (179 ± 8), (144 ± 17) perfusion unit, with t values respectively 3.71, 5.57, 2.77, P < 0.05 or P < 0.01]. (3) The difference in the overall microvessel density in the wound of rats between two groups was significant (F = 33.25, P < 0.05). On PTD 3, the microvessel density in the wound of rats in group NP was (80 ± 12) per 100-time visual field, which was significantly higher than that of group C[(38 ± 4) per 100-time visual field, t = 9.257, P < 0.05]. On PTD 7, the microvessel density in the wound of rats in two groups were close (t = 1.159, P > 0.05), but the vessels in group NP were regularly arranged with spacious lumen, while the vessels in group C were disorderly arranged with narrow lumen. (4) On PTD 1, 3, mRNA expression levels of VEGF, Fit-1, and Ang-1 in group NP were obviously higher than those in group C (with t values from 1.28 to 11.60, P values all below 0.01). On PTD 7, the mRNA expression level of Ang-1 (27.59 ± 3.55) in group NP was obviously higher than that in group C (19.87 ± 1.86, t = 7.23, P < 0.001), while the mRNA level of its antagonist Ang-2 (5.79 ± 0.61) in group NP was obviously lower than that in group C (17.62 ± 0.85, t = 19.88, P < 0.001). On PTD 3, 7, 14, mRNA levels of Tie-2 in group NP were obviously lower than those in group C (with t values from 8.92 to 15.60, P values all below 0.01).</p><p><b>CONCLUSIONS</b>Negative pressure wound therapy may promote wound angiogenesis by enhancing the expression of Ang-1 and lowering the expression of Ang-2 in diabetic rats.</p>

Co-occlusion of foreign protein into polyhedra with BmNPV polyhedrin / 病毒学报

In order to make clear the packing mechanism of the BmNPV polyhedra, a polyhedrin gene negative recombinant baculovirus, vBmBac(polh-)-5B-EGFP, expressing EGFP was constructed, and used to infect BmN cells jointly with wild-type BmNPV. Fluorescent microscopic observation demonstrated that EGFP and polyhedrin were expressed simultaneously, and the EGFP expression and polyhedra formation occurred in most of the jointly infected cells. Analysis of the purified polyhedra from jointly infected BmN cells showed that the foreign proteins were present in the polyhedra. The results indicated that BmNPV polyhedrin could incorporate proteins other than viral proteins into the polyhedra. It implies that a nonspecific recognition mechanism exists in the embedment of BmNPV polyhedra.

Biological effects of paracrine from insulin stimulated adipose-derived stem cells (ADSC) on human vascular endothelial cells / 中华烧伤杂志

<p><b>OBJECTIVE</b>To study the biological effects of the paracrine from ADSC after being stimulated by insulin on vascular endothelial cells.</p><p><b>METHODS</b>(1) ADSC was isolated from human adipose tissue and cultured in vitro. The third generation cells were collected and divided into insulin group (I, cultured with serum-free DMEM containing 1 x 10(-7) mol/L insulin) and control group (C, cultured with serum-free DMEM) according to the random number table, with 6 slots in each group. Three days later, ADSC culture medium (ADSC-CM) was collected for determination of levels of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) by ELISA. (2) Human umbilical vein endothelial cells (HUVEC) were cultured to the third generation, and they were cultured with special nutrient solution and divided into ADSC-CM with insulin stimulation group (AI), ADSC-CM without insulin stimulation group (AC), insulin group (I, with same concentration as above), blank control group (BC) according to the random number table. Three days later, proliferation of HUVEC was determined with MTT method (with expression of absorbance value). Another two samples of HUVEC were respectively divided into 4 groups as above for determination of apoptosis rate with Annexin V/FITC double-staining 12 hours after culture, and HUVEC migration with scratch adhesion test at post scratch hour (PSH) 12, 24, 36, 48. Data were processed with t test.</p><p><b>RESULTS</b>(1) Compared with those in C group [(287 +/- 47), (577 +/- 84) pg/mL, respectively], the secretion levels of VEGF and HGF in I group [(643 +/- 64), (930 +/- 68) pg/mL, respectively] were significantly increased (with t value respectively 18.869, 18.475, P values all below 0.05). (2) The absorbance value of HUVEC in AI and AC groups was 0.847 +/- 0.042, 0.798 +/- 0.022, respectively, which were higher than that in I and BC groups [0.665 +/- 0.028 (with t value respectively 4.579, 3.732), 0.674 +/- 0.031 (with t value respectively 3.761, 4.073), P values all below 0.01], and that in AI group was higher than that in AC group (t = 2.576, P < 0.05). The apoptosis rates of HUVEC in AI and AC groups [(5.8 +/- 1.9)%, (9.0 +/- 2.0)%, respectively] were obviously lower as compared with that in I and BC groups [(30.4 +/- 6.0)% (with t value respectively 12.891, 10.417), (31.4 +/- 7.4)% (with t value respectively 11.474, 9.783), P values all below 0.05 ], and that in AC group was higher than that in AI group (t = 8.548, P < 0.05). The distance of migration of HUVEC in AI and AC groups were greater than that in I and BC groups at PSH 36, 48, and that in AI group was greater as compared with that in AC group (with t value respectively 4.076, 4.573, P values all below 0.05).</p><p><b>CONCLUSIONS</b>Paracrine from ADSC after being stimulated by insulin can promote proliferation and migration of HUVEC, and suppress its apoptosis, and it is beneficial for tissue vascularization.</p>

Study on colloidal gold labeled anti-buprenorphine monoclonal antibody for rapid test kit to detect buprenorphine / 法医学杂志

OBJECTIVE@#To develope an easy to use, rapid and accurate test for detecting buprenorphine based on the principle of competitive immunoassay.@*METHODS@#Monoclonal antibody against buprenorphine was conjugated with colloidal gold and dispensed on the glass fiber. The complete antigen Buprenorphine-BSA and the goat anti-mouse IgG polyclonal antibody were separately sprayed on the nitrocellulose membrane as the test line (T line) and the control line (C line). The rapid test kit was the final assembled product of test strip with the plastic cover.@*RESULTS@#A total of 100 urine samples were tested for buprenorphine by immunochromatographic and GC/ MS methods. The accuracy was 99.0%. It is found the test kit can only detect by cross reaction with other 45 kind drugs.@*CONCLUSION@#Rapid test kit can detect buprenorphine in the samples in 5 minutes. The cut-off value of the test is 100 ng/mL.