Lowered expression of CCN5 in endometriotic tissues promotes proliferation, migration and invasion of endometrial stromal cells / 南方医科大学学报

OBJECTIVE@#To explore the expression of CCN5 in endometriotic tissues and its impact on proliferation, migration and invasion of human endometrial stromal cells (HESCs).@*METHODS@#We collected ovarian endometriosis samples from 20 women receiving laparoscopic surgery and eutopic endometrium samples from 15 women undergoing IVF-ET for comparison of CCN5 expression. Cultured HESCs were transfected with a recombinant adenovirus Ad-CCN5 for CCN5 overexpression or with a CCN5-specific siRNA for knocking down CCN5 expression, and the changes of cell proliferation, migration and invasion were evaluated using CCK-8 assay, wound healing assay and Transwell chamber assay. RT-qPCR and Western blotting were used to examine the expression levels of epithelial-mesenchymal transition (EMT) markers including E-cadherin, N-cadherin, Snail-1 and vimentin in HESCs with CCN5 overexpression or knockdown.@*RESULTS@#CCN5 expression was significantly decreased in ovarian endometriosis tissues as compared with eutopic endometrium samples (P < 0.01). CCN5 overexpression obviously inhibited the proliferation, migration and invasion of HESCs, significantly increased the expression of E-cadherin and decreased the expressions of N-cadherin, Snail-1 and vimentin (P < 0.01). CCN5 knockdown significantly enhanced the proliferation, migration and invasion of HESCs and produced opposite effects on the expressions of E-cadherin, N-cadherin, Snail-1 and vimentin (P < 0.01).@*CONCLUSION@#CCN5 can regulate the proliferation, migration and invasion of HESCs and thus plays an important role in EMT of HESCs, suggesting the potential of CCN5 as a therapeutic target for endometriosis.

Effects of dendritic epidermal T cells on proliferation and apoptosis of epidermal cells in wound margin of mice / 中华烧伤杂志

Objective@#To explore the effects of dendritic epidermal T cells (DETC) on proliferation and apoptosis of epidermal cells in wound margin of mice and its effects on wound healing.@*Methods@#Twenty-eight healthy specific pathogen free (SPF) C57BL/6 wild-type (WT) male mice aged 8-12 weeks and 60 SPF T lymphocyte receptor δ-knockout (TCR δ-/-) male mice aged 8-12 weeks were selected to conduct the following experiments. (1) Eight WT mice were selected to isolate epidermal cells and primarily culture DETC according to the random number table. Morphological observation and purity identification of DETC by flow cytometer were detected immediately after culture and on culture day (CD) 15 and 30, respectively. (2) According to the random number table, 5 WT mice and 5 TCR δ-/- mice were selected and enrolled into WT control group and TCR δ-/- group. Round full-thickness skin defect with diameter of 6 mm was made on the back of each mouse. The wound healing condition was observed immediately after injury and on post injury day (PID) 2, 4, 6, 8, 10, and the percentage of residual wound area was calculated. (3) Mice were selected to group and reproduce model of full-thickness skin defect as in experiment (2). On PID 3, the tissue of wound margin was collected for hematoxylin eosin staining, and the length of new epithelium was measured. (4) Mice were selected to group and reproduce model of full-thickness skin defect as in experiment (2). On PID 3, epidermal tissue of wound margin was collected to determine expression of proliferating cell nuclear antigen (PCNA) using Western blotting for evaluation of proliferation of epidermal cell. (5) Mice were selected to group and reproduce model of full-thickness skin defect as in experiment (2). On PID 3, epidermal tissue of wound margin was selected and digested into single-cell suspension, and apoptosis of cells was detected by flow cytometer. (6) Forty TCR δ-/- mice were selected to carry out the same treatment as in experiments (2)-(5). According to the random number table, these mice were enrolled into TCR δ-/- control group and TCR δ-/-+ DETC group, with 5 mice in each group for each experiment. Round full-thickness skin defect was made on the back of each mouse. DETC in the number of 1×105 (dissolution in 100 μL phosphate with buffer purity above 90%) were injected through multiple points of wound margin of mice in TCR δ-/-+ DETC group immediately after injury, and equal volume of phosphate buffer was injected into mice of TCR δ-/- control group with the same method as above. Data were processed with one-way analysis of variance for repeated measurement, t test, and Bonferroni correction.@*Results@#(1) Along with the culture time elapse, the number of dendritic structures of DETC increased gradually. The percentage of T lymphocytes was 4.67% and 94.1% of these T lymphocytes were DETC. The purity of DETC on CD 15 was 18.50% and the purity of DETC on CD 30 was 98.70%. (2) Immediately after injury, the wound healing condition of mice in WT control group and TCR δ-/- group was similar. The wound healing speed of mice in TCR δ-/- group was slower than that in WT control group on PID 2-10. The percentages of residual wound area of mice in TCR δ-/- group on PID 2, 4, 6, 8, and 10 were increased significantly compared with those in WT control group (t=3.492, 4.425, 4.170, 4.780, 7.318, P<0.01). (3) The length of new epithelium of mice in TCR δ-/- group on PID 3 was (359 ± 15) μm, which was obviously shorter than that in WT control group [(462±26) μm, t=3.462, P<0.01]. (4) Immediately after injury, wound condition of mice in TCR δ-/-+ DETC group and TCR δ-/- control group was similar. Compared with TCR δ-/-+ DETC group, the wound healing speed of mice in TCR δ-/- control group were obviously slower on PID 2-10. The percentages of residual wound area of mice in TCR δ-/-+ DETC group on PID 2, 4, 6, 8, and 10 were decreased significantly compared with those in TCR δ-/- control group (t=2.308, 3.725, 2.698, 3.707, 6.093, P<0.05 or P<0.01). (5) On PID 3, the length of new epithelium of mice in TCR δ-/-+ DETC group was (465±31) μm, which was obviously longer than that in TCR δ-/- control group [(375±21) μm, t=2.390, P<0.05]. (6) On PID 3, PCNA expression of epidermal cell in wound margin of mice in TCR δ-/- group was 1.25±0.04, which was obviously lower than that in WT control group (2.01±0.09, t=7.415, P<0.01). (7) On PID 3, PCNA expression of epidermal cell in wound margin of mice in TCR δ-/-+ DETC group was 1.62±0.08, which was significantly higher than that in TCR δ-/- control group (1.05±0.14, t=3.561, P<0.05). (8) On PID 3, apoptosis rate of epidermal cell in wound margin of mice in TCR δ-/- group was (16.1±1.4)%, which was higher than that in WT control group [(8.1±0.6)%, t=5.363, P<0.01]. (9) On PID 3, apoptosis rate of epidermal cell in wound margin of mice in TCR δ-/-+ DETC group was (11.4±1.0)%, which was obviously lower than that in TCR δ-/- control group [(15.4±1.4)%, t=2.377, P<0.05].@*Conclusions@#DETC participates in the process of wound healing though promoting the proliferation of epidermal cells in wound margin and inhibit the apoptosis of these cells.

Metabolic characteristics of active parts of lipid-lowering flavonoid extract of Daidai in liver and intestinal microsomes of rats / 中国中药杂志

The paper studies and compares the metabolic difference of active ingredients of lipid-lowering flavonoid extract of Daidai in rat livers and intestinal microsomes,in order to explore the phase Ⅰ metabolism characteristics of active ingredients in livers and intestines. UPLC-MS/MS was used to establish a quantitative analysis method for active ingredients,neohesperidin and narngin,in a phase Ⅰ metabolism incubation system of liver and intestinal microsomes. Differential centrifugation was used to make liver and intestinal microsomes of rats. A phase Ⅰ metabolism incubation system was established,and the concentrations of the residual at different incubation time points were analyzed. Graphs were plotted to calculate the metabolic elimination half-life of the main active parts,with the natural logarithm residual percentage values ln( X) at different time points as the y axis,and time t as the x axis. The metabolism characteristics of the active ingredients were compared. The established UPLC-MS/MS quantitative analysis method has a good specialization,standard curve and linear range,accuracy and precision,with a satisfactory lower quantitative limit. The method allows quantitative detection of the active ingredients in a phase Ⅰ metabolism incubation system of liver and intestinal microsomes of rats. In the rats liver microsomes incubation system,the metabolic elimination half-life of neohesperidin and narngin were( 2. 20 ± 0. 28) h and( 1. 97±0. 28) h respectively. The elimination half-life of neohesperidin was larger than that of narngin,but with no statistically significant difference. In the rats intestinal microsomes incubation system,the metabolic elimination half-lives of neohesperidin and narngin were( 3. 68±0. 54) h and( 2. 26±0. 13) h respectively. The elimination half-life of neohesperidin was larger than that of narngin,with statistically significant differences( P<0. 05). The elimination half-lives of the active ingredients in liver microsomes were smaller than those in intestinal microsomes. The experiment results showed that the active ingredients of lipid-lowering flavonoid extract of Daidai had different elimination half-lives in phase Ⅰ rats liver and intestinal microsomes incubation system. This implied that they had different metabolic characteristics in rats liver and intestine,and liver may be the main metabolism site of the active ingredients. The phaseⅠ metabolism of narngin was stronger than that of neohesperidin. The differences between their metabolic characteristics may be related to the binding sites of B-ring hydroxyl in flavonoid glycosides and the number of methoxyl group. The results provided an important experimental basis for further development and clinical application of lipid-lowering flavonoid extract preparation of Daidai.

Binding characteristics of plasma protein in active parts of Daidai lipid-lowering flavonoid extract / 中国中药杂志

To study the binding capacity of active ingredients of Daidai lipid-lowering flavonoid extract and plasma protein,investigate the ways to improve the traditional formula for calculating protein binding rates based on ultrafiltration,and increase the stability and reliability of the experimental results. UPLC-MS/MS was used to establish a quantitative analysis method for simultaneous determination of active ingredients( neohesperidin and narngin) in ultrafiltrate. The protein binding rates were calculated by the traditional ultrafiltration formula. The correction factors( F) were introduced later,and the binding rates calculated with the correction factors were compared with those without the correction factors. The binding capacity of the extract and plasma protein was evaluated. The quantitative analysis method established by UPLC-MS/MS had a good specificity. The standard curve and linear range,method accuracy,precision and lower limit of quantitation all met the requirements. The method met the requirement for quantitative detection of the active ingredients in ultrafiltrate after the rat plasma was filtrated in the ultrafiltration tube. Under the experimental conditions,the binding rates of both active ingredients( neohesperidin and narngin) were higher than 90%. The active ingredients and rat plasma protein were bound in a concentration-dependent manner,with statistically significant differences( P<0. 01). There was no statistically significant difference between the protein binding abilities of the two active ingredients with rat plasma protein. Therefore,the active ingredients of Daidai lipid-lowering flavonoid extract had a relatively strong binding strength with rat plasma protein,and they were bound in a concentration-dependent manner. Additionally,when calculating protein binding rates by the traditional ultrafiltration formula,the correction factors could be introduced to effectively reflect the errors of multiple ingredient groups in traditional Chinese medicine extracts.This correction method could provide a reference thinking and practical reference for the improvement of the determination method of the traditional Chinese medicine plasma protein binding ability based on ultrafiltration.

Testosterone Undecanoate Pills improves insulin resistance in type-2 diabetes men with hypogonadism / 中华男科学杂志

Objective@#To evaluate the effects of Testosterone Undecanoate Pills (TUP) on insulin resistance (IR) in type-2 diabetes men with hypogonadism.@*METHODS@#We randomly divided 82 type-2 diabetes patients with hypogonadism into a treatment (n = 42) and a control group (n = 40), both maintaining their glucose- and lipid-reducing therapies, while the former treated orally with TUP in addition. After 6 months of medication, we compared the body mass index (BMI), waist circumference (WC), blood glucose level, HbA1c, lipid profile, IR index obtained by homeostatic model assessment (HOMA-IR), insulin sensitivity index (ISI), sex hormone levels, and sexual function scores between the two groups of patients.@*RESULTS@#Compared with the baseline, the patients in the treatment group showed significant decreases after medication in BMI (［26.71 ± 2.39］ vs ［25.15 ± 2.28］ kg/m2, P 0.05).@*CONCLUSIONS@#TUP can significantly improve insulin resistance in type-2 diabetes men with hypogonadism.

The mediating effects of perceived social support between clinicians' work pressure and compassion fatigue / 中华行为医学与脑科学杂志

Objective To explore the relationship among clinicians' work pressure,perceived social support and compassion fatigue,and to provide a new prospective for the localization studying of intervention and treatment to compassion fatigue.Methods Data of scale for occupational stressors on clinician,perceived social support scale and professional quality of life scale were collected from a sample of 533 clinicians and analyzed with structural equation modeling to study the relationship among clinicians' work pressure,perceived social support and compassion fatigue.Results (1)Work pressure(2.40±0.45),burnout (2.14±0.54) and secondary traumatic stress(1.93±0.60) scores of the clinicians with high perceived social support were significantly lower than that of the clinicians with low perceived social support (2.78±3.67,2.73± 0.59,2.32±0.71;t=7.68,-9.44,8.77,5.07;P＜0.01).Compassion satisfaction scores (4.15±0.63) of the clinicians with high perceived social support were significantly higher than that of the clinicians with low perceived social support (3.40±0.71,t =-9.44,P＜0.01).(2) According to relevant results,work pressure was significantly negative correlation with both perceived social support (r=-0.34,P＜0.01) and compassion satisfaction (r=-0.44,P＜0.01),and significantly positive correlation with both burnout (r=0.69,P＜0.01) and secondary traumatic stress(r=0.53,P＜0.01);while perceived social support was significantly positive correlation with compassion satisfaction (r=0.42,P＜0.01),and significantly negative correlation with burnout (r=-0.40,P＜0.01) and secondary traumatic stress(r=-0.26,P＜0.01).(3) According to the results of structural equation modeling,perceived social support played a partly mediating role in the effect of work pressure toward compassion satisfaction and fatigue with the intermediary effect of 55.4％ and 12.5％.Conclusion Perceived social support plays a mediation role between work pressure and compassion fatigue for clinicians,and better social support of the clinician is beneficial to clinicians mental health level.

Uterine macrophages affect embryo implantation via regulating vascular endothelial growth factor A in mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of endometrial macrophages in embryo implantation and in regulating the expression of vascular endothelial growth factor A (VEGFA) in mouse endometrium during the peri-implantation period.</p><p><b>METHOD</b>At D3.5 (D0.5 defined as the morning when a vaginal plug was observed), pregnant mice were divided randomly into experimental group, control group and blank group. In the experimental group, the mice were subjected to intrauterine injection of clodronate liposomes on the left side of uterus to eliminate the macrophages, and PBS liposomes on the right side. PBS liposomes and PBS were administered in the control and blank groups, respectively. The uterine tissues were collected on D5.5 and stained with trypan blue to show the implantation sites. Flow cytometry was performed to examine the percentage of F4/80(+) CD11b(+) macrophages macrophages in the uterus. F4/80(+) macrophage population within the endometrium and ovary and changes in VEGFA expression at the implantation and non-implantation sites were examined using immunohistochemistry.</p><p><b>RESULTS</b>Endometrial F4/80(+) CD11b(+) macrophages macrophages were significantly reduced by 74% following intrauterine injection of clodronate liposomes (P<0.05). The number of macrophages in the ovaries showed no significant difference among the 3 groups. In the experimental group, the left side of the uterine showed imcomplete cavity closure with a lower number of implantation site than the right side (2.20∓1.81 vs 5.10∓1.91, P<0.05). VEGFA expression at the implantation site were significantly decreased in the endometrium on the left side with macrophage suppression as compared with that on the right side (P<0.05).</p><p><b>CONCLUSION</b>Endometrial macrophages appear to modulate uterine receptivity by regulating the expression of VEGFA to affect embryo implantation, suggesting the important role of macrophages in embryo implantation.</p>

Media of rat macrophage NR8383 cells with prostaglandins E2-induced VEGF over-expression promotes migration and tube formation of human umbilical vein endothelial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of prostaglandins E2 (PGE2) in enhancing vascular endothelial growth factor (VEGF) expression in a rat macrophage cell line and the effect of the media from PGE2-inuced rat macrophages on angiogenetic ability of human umbilical vein endothelial cells (HUVECs) in vitro.</p><p><b>METHODS</b>Western blotting and qPCR were employed to investigate the expressions of VEGF protein and mRNAs in rat macrophage cell line NR8383 stimulated by PGE2 in the presence or absence of EP2 receptor inhibitor (AH6809) and EP4 receptor inhibitor (AH23848). Conditioned supernatants were obtained from different NR8383 subsets to stimulate HUVECs, and the tube formation ability and migration of the HUVECs were assessed with Transwell assay.</p><p><b>RESULTS</b>PGE2 stimulation significantly enhanced the expression of VEGF protein and mRNAs in NR8383 cells in a dose-dependent manner. The supernatants from NR8383 cells stimulated by PGE2 significantly enhanced tube formation ability of HUVECs (P<0.05) and promoted the cell migration. Such effects of PGE2 were blocked by the application of AH6809 and AH23848.</p><p><b>CONCLUSION</b>PGE2 can dose-dependently increase VEGF expression in NR8383 cells, and the supernatants derived from PGE2-stimulated NR8383 cells can induce HUVEC migration and accelerate the growth of tube like structures. PGE2 are essential to corpus luteum formation by stimulating macrophages to induce angiogenesis through EP2/EP4.</p>

Comparative study on Argus and artificial methods for MRI scanning of femoral head necrosis area / 医疗卫生装备

Objective To explore the advantages of Argus method by comparing the accuracy and timeliness of Argus and artificial methods for measuring femoral head necrosis area in MRI scanning.Methods Totally 17 patients (31 hips) were measured with Argus and artificial methods respectively for the necrosis area, and then the measuring results and time were compared, and the correlation was investigated between the results and the patients' pain degree, along with that between the results and the extent of femoral head collapse.Results The necrosis area ratios determined by Argus and artificial methods were (33.5±4.08)%and (34.6±4.06)%respectively, with no statistical difference between the ratios (P>0.05). The time consumed by artificial method was (21.3 ±3.62)min, significantly longer than (7.89 ±1.03)min by Argus method, with P<0.001. Regression analysis proved that the necrosis areas were positively correlated with the patients' pain degree, and the correlation coefficient by Argus method was 0.807 8, more than 0.740 9 by artificial method. The femoral heads of 11 cases(16 hips) collapsed in the follow-up period, the necrosis areas were positively correlated with the patients collapse level, but the correlation coefficient by Argus method was 0.783 8, more than 0.726 7 by artificial method.Conclusion Argus method gains high accuracy and timeliness when used in MRI scanning of femoral head necrosis area, and thus is worth popularizing clinically.

Intracytoplasmic injection of morphologically selected spermatozoa from patients with male factor infertility: clinical and embryo development outcomes / 南方医科大学学报

<p><b>OBJECTIVE</b>To test whether intracytoplasmic injection of morphologically selected spermatozoa (IMSI) from patients with male factor infertility can improve the clinical and embryo development outcomes of intracytoplasmic sperm injection-embryo transfer (ICSI-ET).</p><p><b>METHODS</b>We performed IMSI for 82 couples diagnosed with obstructive azoospermia at high magnification (×6600) and traditional ICSI for another 91 couples using testicular sperms. We also performed IMSI for 44 couples with teratozoospermia at high magnification (×6600) and traditional ICSI for 71 patients using ejaculated sperms. The clinical and embryo development outcomes were compared between the cycles.</p><p><b>RESULTS</b>For obstructive azoospermia, IMSI and ICSI showed no significant difference in the rates of cleavage (95.5% vs 96.7%), D3 top quality embryos (28.2% vs 29.2%), implantation (26.4% vs 32.3%), pregnancy (47.3% vs 50%), blastocyst formation (54.3% vs 54.6%), or abortion (14% vs 7.3%) (P>0.05), but a significantly higher normal fertilization rate was achieved in IMSI group (84.3% vs 77%, P<0.05). For teratozoospermia, the 2 techniques resulted in no significant differences in the rates of cleavage (96.2% vs 95.2%), D3 top quality embryo (27.6% vs 27.1%), implantation (28.2% vs 30.7%), pregnancy (43.7% vs 43.2%), or abortion (9.7% vs 10.5%) (P>0.05), but the normal fertilization rate (68% vs 75.5%) and the blastocyst formation rate (54.6% vs 67.9% ) were significantly higher in IMSI group (P<0.05).</p><p><b>CONCLUSION</b>IMSI can improve the normal fertilization rates in couples with male factor infertility (including obstructive azoospermia and teratozoospermia) and increase blastocyst formation rate in cases of azoospermia.</p>

Protective effect of α-melanocyte stimulating hormone on glutamate-induced retinal excitotoxicity / 中华实验眼科杂志

Background The excitotoxicity to retinal neurons caused by abnormal elevation of glutamate in retina is a common pathology concomitant with major blind-causing eye diseases.However,an effective approach to protect retinal neurons from glutamate-induced excitotoxicity is still lack.Intraperitoneal administration of α-melanocyte stimulating hormone(α-MSH)has been shown to protect hippocampal neurons from glutamate-induced excitotoxicity.Objective This study was to investigate the protective effect of α-MSH on glutamate-induced excitotoxicity in a chicken embryonic retinal explant culture system.Methods The retinas were isolated from chick embryos at embryonic day 9(E9) and cultured as explants.The explants at 3,5 and 7 days in vitro and the retinas at corresponding embryonic day 12,14 and 16(E12,E14,E16)were collected.The morphology of explant cultures was examined by hematoxylin and eosin staining,and the expression of melanocortin receptors (MCRs)was analyzed by real-time PCR.In the experiment of glutamate-induced excitotoxicity,the retinal explants at 4 days in vitro were treated with glutamate for 48 hours,α-MSH was incubated with the explants 30 minutes before and during the glutamate treatment period.Then the apoptotic cells were detected by TUNEL staining and quantified.The glutamate alone treated-explants and those treated with culture media were included as controls.The expression of glial fibrillary acidic protein(GFAP) at 48 hours after treatment in all retinal explants was analyzed by real-time PCR.Results Hematoxylin and eosin staining showed that the retinal explants exhibited similar morphology to those observed in the retinas from chick embryos at the corresponding developmental stages.The real-time PCR analyses of chick retinas showed that MC1R mRNA level at E9,E12,E14 and E16 was significantly lower than that in post-hatch day 1 (all P=0.000) ;whereas the transcript level of MC5R was significantly increased from E9 to E12 and E14 (both P =0.000),and then gradually decreased from E14 to P1.The expression of these genes showed similar temporal patterns in the retinal explant cultures.TUNEL staining revealed that treatment of the retinal explant cultures with α-MSH substantially and significantly reduced number of apoptotic cells induced by glutamate (P =0.000),which was accompanied by significant suppression of glutamate-induced GFAP up-regulation (P =0.000).Conclusions Application of α-MSH dramatically ameliorated glutamate-induced cell death in retinal explant cultures.This protective effect may be due to α-MSH-mediated suppression of astrogliosis caused by abnormal elevation of glutamate.

Study on the psychosocial status and related factors in three universities during severe acute respiratory syndrome epidemic in Beijing / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the psychosocial status and related factors among university students during severe acute respiratory syndrome (SARS) epidemic in Beijing.</p><p><b>METHODS</b>By means of stratified cluster sampling, symptom checklist-90 (SCL-90) and questionnaire on general information were applied among 6800 students in three universities in Beijing.</p><p><b>RESULTS</b>There were 6280 valid questionnaires gathered. In order to control and prevent SARS, strict management was conducted in three universities which providing various social supports. Out of the 6280 students, 460 had SCL-90 positive symptoms with a rate of 7.3%. Risk factors of SCL-90 positive symptom were found as follows: major in arts (OR = 2.00), misconception on the control and prevention of SARS (OR = 1.91), considering measures non-effective (OR = 2.25), and do not believe that SARS can be under control (OR = 3.57). Protective factors of SCL-90 positive symptom would include as: being female (OR = 0.77), being graduate students (OR = 0.38), and being not much influenced on study and daily life during the period of strict management (OR = 0.54).</p><p><b>CONCLUSION</b>Psychosocial status of students was influenced by their knowledge and attitude on SARS. Various social supports might keep the university students to having a healthy psychosocial status.</p>