Protective Effect of Ginsenosides Rg1 and Rb1 Against Intestinal Epithelial Barrier Injury Induced by Lipopolysaccharide in Vitro / 中国实验方剂学杂志

ObjectiveTo investigate the effects of ginsenoside Rg1 and ginsenoside Rb1 on the release of inflammatory factors of human myeloid leukemia monocytes （THP-1） induced by lipopolysaccharide （LPS） and their protective effects on the inflammatory injury of intestinal epithelial cells （Caco-2） induced by THP-1 cell activation based on the co-culture system of THP-1 and Caco-2. MethodFirstly，the microfluidic chip of co-culture of THP-1 and Caco-2 cells was prepared. In the experiment， a blank group， an LPS group， and drug intervention groups were set up.The cells in the blank group were cultured conventionally. In the LPS group，LPS （1 mg·L-1） was added to the lower THP-1 cells after the upper Caco-2 cells formed a monolayer barrier. On the basis of the LPS group， 33 mg·L-1 ginsenoside Rg1 and 33 mg·L-1 ginsenoside Rb1 were added to THP-1 cells respectively. After the co-culture of THP-1 cells and Caco-2 cells for 24 hours， the fluorescein isothiocyanate （FITC）-Dextran fluorescence value in the lower chip channel was detected by FITC-Dextran tracer method. A blank group， an LPS group，and drug intervention groups were set up in the THP-1 cell experiment. THP-1 cells in the blank group were cultured conventionally. In the LPS group， LPS （1 mg·L-1） was added to THP-1 cells.Ginsenoside Rg1 and ginsenoside Rb1 of the corresponding doses （11，33，100 mg·L-1） were added to the drug intervention groups respectively on the basis of the LSP group. After 24 hours of cell culture， the activity of THP-1 cells was detected by cell counting kit-8 （CCK-8）. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the expression of inflammatory cytokines such as interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and tumor necrosis factor （TNF）-α of THP-1 cells. A blank group， an LPS group， and drug intervention groups were set up in the Caco-2 cell experiment. Caco-2 cells in the blank group were cultured conventionally， and in other groups， the corresponding cell supernatant in the second part of the THP-1 cell experiment was employed in Caco-2 cells. After 24 hours of cell culture，the activity of Caco-2 cells was detected by CCK-8. Real-time PCR was used to detect the expression of IL-6，interleukin-8 （IL-8）， TNF-α， and Occludin of Caco-2 cells. The expression of tight junction protein Occludin in Caco-2 cells was detected by Western blot. ResultBoth ginsenoside Rg1 and ginsenoside Rb1 could effectively protect LPS-induced intestinal epithelial barrier permeability in the co-culture system of THP-1 and Caco-2 cells （P<0.01）. Ginsenosides Rg1 and Rb1 antagonized LPS-induced increased expression of IL-6，IL-1β， and TNF-α in THP-1 cells （P<0.05）. When the supernatant of THP-1 cells treated with ginsenosides Rg1 and Rb1 was co-cultured with Caco-2 cells， the expression of IL-6，IL-8， and TNF-α in Caco-2 cells was significantly reduced （P<0.01）， and the expression of tight junction protein Occludin was up-regulated. ConclusionIn the co-culture system of THP-1 and Caco-2 cells simulating the intestinal epithelial barrier function in vitro，ginsenosides Rg1 and Rb1 play a protective role against LPS-induced intestinal epithelial barrier injury by regulating the release of inflammatory cytokines by THP-1 cells， thereby regulating the inflammatory response and cell barrier integrity of Caco-2 cells.

Effect of Euphorbiae Ebarcteolatae Radix Bacteriostasis Ointment on Chronic Eczema in Mice / 中国实验方剂学杂志

Objective:To observe the anti-inflammatory effect and its mechanism of Euphorbiae Ebarcteolatae Radix bacteriostasis ointment in eczema mice induced by 2,4-dinitrochlorobenzene (DNCB). Method:A total of 40 ICR adult mice were randomly divided into control group，model group，hydrocortisone Butyrate cream group (0.09g·kg-1) and Euphorbiae Ebarcteolatae Radix bacteriostasis ointment group (0.09 g·kg-1), with 10 mice in each group. Except for the normal group, other groups were given DNCB to induce the chronic eczema model. Twenty-four hours after DNCB stimulation, they were given the corresponding drugs through auricle and back, twice a day for 10 days. After drug intervention, efforts were made to measure the change of thickness and weight of the middle ear, assess the allergic effect, and calculate the spleen index of the mice. Optical microscope was used to observe the pathological changes in ear tissues of mice. And the levels of serum interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), interleukin-4 (IL-4) in mice were determined by multiplex immunoassay. Result:Compared with control group, the thickness and weight of right ears, score of allergic effect, spleen index and the levels of IL-2 and IL-4 in serum showed significant increases in model group (P<0.05，P<0.01). The histopathology injuries of ear were aggravated. Compared with model control group, Euphorbiae Ebarcteolatae Radix bacteriostasis ointment could reduce ear thickness and score of allergic effect, regulate the spleen index, decrease the inflammation factor in serum such as IL-2 and IL-4 (P<0.05，P<0.01), and improve ear histopathology injuries. Conclusion:Euphorbiae Ebarcteolatae Radix bacteriostasis ointment may have a good effect on eczema.

Analysis of Non-genetic Factors Influencing the Overall Survival of Multiple Myeloma Patients / 中国实验血液学杂志

OBJECTIVE@#To explore the non-genetic factors of overall survival in patients with multiple myeloma (MM).@*METHODS@#Kaplan-Meier survival curve, Log-rank test and Cox regression model were used to carry out univariate and multivariate retrospective analysis on clinical and laboratory parameters of 51 patients who were newly diagnosed with MM and had complete follow-up data in the Department of Hematology, the Affiliated Jiangning Hospital of Nanjing Medical University from November 2011 to October 2019.@*RESULTS@#Fifty-one patients included 29 males and 22 females. Followed up to December 2019, 21 cases died and 30 cases survived. The univariate analysis showed that the overall survival time of the patients was influenced by age, disease stage, standard treatment, new drugs, maintenance treatment, hypercalcemia, globulin, albumin, and hemoglobin. The overall survival time of patients with age <65 years old, ISS stage I and II, standardized treatment, new drugs, normal or below normal blood calcium, normal or below normal globulin, albumin ≥35 g/L or hemoglobin ≥100 g/L was prolonged significantly (P<0.05). The multivariate analysis showed that maintenance treatment, hypercalcemia (≥2.6 mmol/L), and hemoglobin (<100 g/L) were independent risk factors influencing the prognosis of MM patients.@*CONCLUSION@#Patients with blood calcium ≥2.6 mmol/L, hemoglobin <100 g/L, and who do not undergo regular maintenance therapy show a poor prognosis.

Decitabine combined with arsenious acid in the treatment of patients with higher-risk myelodysplastic syndromes and chronic myelomonocytic leukemia / 中国实用内科杂志

OBJECTIVE: To evaluate the clinical efficacy of decitabine combined with arsenious acid in the treatment of patients with higher-risk myelodysplastic syndromes(MDS) and chronic myelomonocytic leukemia(CMML). METHODS: Totally 39 patients with MDS and 8 patients with CMML received the treatment of decitabine and arsenious acid from April 2016 to December 2018. Decitabine [20 mg/(m~2·d)] and arsenious acid [0.15 mg/(m~2·d)] were administered intravenously for 5 consecutive days every 4-6 weeks. Patients who achieved complete or partial remission entered into the consolidation cycle. Efficacy and influencing factor were analyzed. RESULTS: Clinical response were observed in 31 patients after a median of 2 courses(ranging 1-12) of treatment. The overall response rate(ORR) was 66.0%. The median duration of response was 16 weeks(ranging 2-52 weeks). There were 8 cases(17.0%) of complete remission(CR), 10 cases(21.3%) of partial remission(PR),12 cases(25.5%) of hematological improvement(HI), 1 case(2.1%) of marrow complete remission(mCR), 8 cases(17.0%) of stable disease(SD), and 1 case(2.1%) of progressive disease(PD). By next generation sequencing, 25 genes mutated with 70 times in 33 cases. The mutation frequency of epigenetic regulators(57.6%) was higher than splicing factors(33.5%), transcription factors and kinase signaling(54.5%),and TP53(21.2%)(P<0.01). There was no significant difference in response rates among these patients(47.4%, 54.5%, 50.0% and85.7%, P=0.977). Gene mutation frequency(VAF) of patients who responded to the regimen declined significantly(16.67% vs. 10.26%,P=0.014). CONCLUSION: Decitabine combined with arsenious acid has significant effect in the treatment of patients with higher-risk MDS and CMML and is well-tolerated. Gene mutation test results by next generation sequencing might be related to clinical response.

The protective effect of astrocyte inhibitor fluorocitrate on subarachnoid hemorrhage in rats / 西安交通大学学报(医学版)

Objective To study the effects of fluorocitrate (FC),astrocytic metabolic inhibitor,on early brain injury after subarachnoid hemorrhage (SAH)in rats so as to explore the mechanism of early brain injury mediated by astrocyte after SAH.Methods Thirty six SD male rats were randomly divided into sham group,SAH 3 d group and SAH 3 d+FC group,with 12 rats in each group.The model of SAH was established by an endovascular perforation technique. FC was injected into the lateral ventricle. Three days after SAH, the expressions of GFAP,Iba-1 and TNF-α were detected using immunohistochemistry in all rats.The expression and phosphorylation of NF-κB in the nucleus were detected by Western blot.The expressions of TNF-α,IL-1βand IL-6 were detected by ELISA.Cell apoptosis was detected by TUNEL.Results Neuronal swelling,nuclear anomalies, disorganization of micrangium,abnormality of endothelial cells appeared in SAH 3 d group,but not in sham group. The expressions of NSE,GFAP and Iba-1 were significantly increased in SAH 3 d group compared with those in sham group (all P＜0.05).The number of apoptotic cells was increased.The expression and phosphorylation of NF-κB were significantly increased.The expressions of TNF-α,IL-1β and IL-6 were significantly increased.However, neuronal injuries were alleviated by injecting FC.The expressions of NSE,GFAP and Iba-1 in SAH 3 d+FC group were inhibited.The number of apoptotic cells in SAH 3 d+FC group was decreased compared with that in SAH 3 d group.The expression and phosphorylation of NF-κB were decreased by FC.The expressions of TNF-α,IL-1β and IL-6 were significantly decreased by FC (P＜0.05).Conclusion Astrocytes may play an important role in early brain injury after SAH,underlying the mechanism that they released TNF-α,IL-1βand IL-6 and activate microglia by activating NF-κB signal.The inhibition of excess activation of astrocytes may plays a protective role in early brain injury after SAH.

Transdermal characteristics and mechanism of asiaticoside nanoemulsions and asiaticoside nanoemulsions-based gels / 中国中药杂志

To prepare the asiaticoside nanoemulsions (ASI-NEs) and asiaticoside nanoemulsions-based gels (ASI-NBGs), compare them with the commercial cream of asiaticoside (ASI-C) in terms of transdermal characteristics, and investigate the transdermal mechanism of ASI-NEs and ASI-NBGs. Their transdermal characteristics were studied by using Franz diffusion cells. The effect of topical ASI-NEs and ASI-NBGs on ultrastructure of rabbit skin was evaluated by using HE staining method. The localization and the permeation pathway of asiaticoside were visually investigated by using laser scanning confocal microscope (CLSM). The transdermal studies in vitro showed that the cumulative amount of ASI permeated from ASI-NEs and ASI-NBGs at 12 h after application were (3 504.30±180.93), (1 187.40±128.88) μg·cm⁻² respectively, 6.57, 2.23 times of that in the control group of ASI-C; the drug deposition of ASI-NEs and ASI-NBGs in skin was (159.48±7.47), (120.53±5.71) μg·cm⁻² respectively, 5.93, 4.48 times of that of ASI-C. HE staining of the rabbit skin after application of ASI-NEs and ASI-NBGs showed that the epidermis structure was basically intact; stratum corneum was loosed and the keratin fragment was increased; at the same time, the gap of prickle cell was increased and the basal cells were arranged loosely. The study of CLSM showed that significant percutaneous enhancer effect was observed for ASI-NEs after the topical application of 6 h, as the fluorescent compound was penetrated in the dermis and diffused uniformly. The fluorescence area and the integral optical density (IOD) were 28.81, 32.51 times of that in the FITC aqueous solution group, respectively. The fluorescent preparations showed strong fluorescence in the epidermis, but weak in deeper layers; with the increase of treatment time, the fluorescence in deeper layer was increased and stronger in skin appendages. The prepared ASI-NEs and ASI-NBGs have good transdermal characteristics and the transdermal mechanism is related to breaking the ultrastructure of stratum corneum and penetrating by the path of skin adnexa.

Serum Gamma-glutamyl Transferase Levels Predict Functional Outcomes after Aneurysmal Subarachnoid Hemorrhage / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>We aim to explore the potential association between serum gamma-glutamyl transferase levels and functional outcome after aneurysmal subarachnoid hemorrhage in a Chinese population.</p><p><b>METHODS</b>A total of 386 aneurysmal subarachnoid hemorrhage patients were included in the study from September 2007 to February 2015. Baseline serum gamma-glutamyl transferase levels and 6-month follow-up functional outcomes were determined. A poor outcome was defined as a modified ranking scale score of ⋝ 3. The multivariable logistic model was used to analyze the relationship between serum gamma-glutamyl transferase and clinical outcomes after aneurysmal subarachnoid hemorrhage.</p><p><b>RESULTS</b>The adjusted poor outcome rates of patients with gamma-glutamyl transferase levels of < 30 U/L, 30-50 U/L and ⋝ 50 U/L were 16.7%, 19.6%, and 34.4%, respectively (P < 0.01). The age-sex and multivariable adjusted odds ratios (95% confidence intervals) of poor prognosis comparing the top group (⋝ 50 U/L) with the lowest group (< 30 U/L) were 5.76 (2.74-12.13), 6.64 (2.05-21.52), and 6.36 (1.92-21.02). A significant linear trend existed between gamma-glutamyl transferase level and aneurysmal subarachnoid hemorrhage prognosis. This association was also observed among nondrinkers.</p><p><b>CONCLUSION</b>Patients with higher gamma-glutamyl transferase levels were more likely to have a poor prognosis. Serum gamma-glutamyl transferase can be considered to be an independent predictor of functional outcomes after aneurysmal subarachnoid hemorrhage.</p>

Biological characteristics of synovial-derived mesenchymal stem cells in serum-free culture media and in vitro construction of three-dimensional cartilage in vitro / 中华整形外科杂志

Objective@#To explore the biological characteristics of synovial fluid-derived mesenchymal stem cells (SF-MSCs) cultured in serum-free medium and the ability of in vitro reconstruction of three-dimensional cartilage combined with scaffold material.@*Methods@#Human SF-MSCs were cultured in serum medium and mesenchymal stem cells medium-serum free (MSCM-sf) respectively, then the proliferative ability and morphology of SF-MSCs were compared; The third passage SF-MSCs cultured in MSCM-sf were identified by flow cytometry, three-way(chondrogenic, osteogenic, adipogenic)differentiation assay and induced for chondrogenic differentiation when combined with polyglycolic acid/polylactic acid (PGA/PLA).@*Results@#SF-MSCs cultured in MSCM-sf had better morphology and proliferative ability than that cultured in serum medium. The expression levels of positive markers of the third passage SF-MSCs cultured in MSCM-sf, such as CD73 (99.5%), CD90 (98.9%) and CD105 (96.5%), were more than 95%. However, the overall negative markers (CD34, HLA-DR and CD11b) expressed less than 2%. Three-way differentiation staining was positive. The combination of SF-MSCs and PGA / PLA can be induced into cartilage in vitro.@*Conclusions@#SF-MSCs cultured in MSCM-sf can be amplified under the condition of maintaining the stem cell characteristics, and can be combined with PGA/PLA scaffold to construct three-dimensional cartilage in vitro.

A Preliminary Study of the Therapeutic Role of Human Early Fetal Aorta-derived Endothelial Progenitor Cells in Inhibiting Carotid Artery Neointimal Hyperplasia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Endothelial cell damage is an important pathophysiological step of restenosis after angioplasty and stenting. Cell transplantation has great therapeutic potential for endothelial recovery. We investigated the effect of transplanting endothelial progenitor cells (EPCs) derived from human early fetal aortas in rat injured arteries.</p><p><b>METHODS</b>The carotid arterial endothelium of Sprague-Dawley rats was damaged by dilatation with a 1.5 F balloon catheter, and then EPCs derived from human early fetal aortas (<14 weeks) were injected into the lumen of the injured artery in transplanted rats, with an equal volume of normal saline injected into control rats. Rats were sacrificed at 2 and 4 weeks after treatment and transplanted cells were identified by immunohistochemical staining with anti-human CD31 and anti-human mitochondria antibodies. Arterial cross-sections were analyzed by pathology, immunohistochemistry, and morphometry.</p><p><b>RESULTS</b>Green fluorescence-labeled EPCs could be seen in the endovascular surface of balloon-injured vessels after transplantation. The intimal area and intimal/medial area ratio were significantly smaller in the transplanted group than in the control (P < 0.05) and the residual lumen area was larger (P < 0.05). After EPC transplantation, a complete vascular endothelial layer was formed, which was positive for human von Willebrand factor after immunohistochemical staining, and immunohistochemical staining revealed many CD31- and mitochondria-positive cells in the re-endothelialized endothelium with EPC transplantation but not control treatment.</p><p><b>CONCLUSION</b>EPCs derived from human early fetal aorta were successfully transplanted into injured vessels and might inhibit neointimal hyperplasia after vascular injury.</p>

Relation of self-esteem, implicit self-esteem and perfectionism in the depressed patients / 中国神经精神疾病杂志

Objective To explore the relationships of self-esteem, implicit self-esteem and perfectionism in the depressed patients. Methods Both of the depression group (n=50) and control group (n=45) completed the self-esteem scale (SES), the self-acceptance questionnaire (SAQ), the Chinese Frost multidimensional perfectionism scale (CFMPS), the implicit association test and the Raven’s Standard Progressive Matrices. The implicit association test were performed before (pre-IAT) and after (later-IAT) the Raven’s Standard Progressive Matrices. Results The SES score was lower in depression group than in control group (P=0.002). To the IAT score, the interaction effect of group and time achieved no statistically different (P=0.735). The group main effect of IAT was significant (P=0.001). The IAT score was higher in de?pression group than in control group (P=0.013). The time main effect was significant (P=0.033). The pre-IAT was higher than later-IAT (P=0.007). The depression group had higher scores of concern over mistakes (P=0.007) and doubt about action (P=0.006) dimension of CFMPS, and lower score of parental expectations (P=0.038) dimension than the control group. The later-IAT was negatively related with parental expectations dimension and the total score of CFMPS in the de?pression group (P<0.05). Depressed patients’SES and SAQ scores were negatively related with concern over mistakes and doubt about action dimension and the total score of CFMPS (P<0.05). Conclusion The instability of implicit self-es?teem of depressed patients is associated with negative perfectionism.

Effects of mild hypothermia on the ROS and expression of caspase-3 mRNA and LC3 of hippocampus nerve cells in rats after cardiopulmonary resuscitation / 世界急诊医学杂志（英文）

@#BACKGROUND: Cardiac arrest (CA) is a common and serious event in emergency medicine. Despite recent improvements in resuscitation techniques, the survival rate of patients with CA is unchanged. The present study was undertaken to observe the effect of mild hypothermia (MH) on the reactive oxygen species (ROS) and the effect of neurological function and related mechanisms. METHODS: Sixty-five healthy male Sprague Dawley (SD) adult rats were randomly (random number) divided into 2 groups: blank control group (n=5) and CPR group (n=60). CA was induced by asphyxia. The surviving rats were randomly (random number) divided into two groups: normothermia CPR group (NT) and hypothermia CPR group (HT). Normothermia of 37 °C was maintained in the NT group after return of spontaneous circulation (ROSC), hypothermal intervention of 32 °C was carried out in the HT group for 4 hours immediately after ROSC. Both the NT and HT groups were then randomly divided into 2 subgroups 12 hours and 24 hours after ROSC (NT-12, NT-24, HT-12, HT-24 subgroups). During observation, the neurological deficit scores (NDSs) was recorded, then the bilateral hippocampi were obtained from rats' head, and monoplast suspension of fresh hippocampus tissue was made immediately to determine the level of intracellular ROS by flow cytometry. Transmission electron microscope was used to observe the ultramicro changes of cellular nucleus and mitochondria. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the expression of caspase-3 mRNA, and western-blotting (WB) was used to determine the level of LC3 in frozen hippocampus tissue. Measured data were analyzed with paired sample t test and One-Way ANOVA. RESULTS: Of 60 rats with CA, 44 (73%) were successfully resuscitated and 33 (55%) survived until the end of the experiment. The NDSs of rats in the NT and HT groups were more significantly reduced than those in the BC group (F=8.107, P<0.05), whereas the NDSs of rats in the HT-12 and HT-24 subgroups were significantly increased in comparison with those NDSs of rats in the NT-12 and NT-24 subgroups, respectively (t=9.692, P<0.001; t=14.374, P<0.001). The ROS in hippocampus nerve cells in the NT and HT groups significantly increased compared to the BC group (F=16.824, P<0.05), whereas the ROS in the HT-12 and HT-24 subgroups significantly reduced compared with that ROS in the NT-12 and NT-24 subgroups, respectively (t=9.836, P<0.001;t=7.499, P<0.001). The expression of caspase-3 mRNA in hippocampus nerve cells in the NT and HT groups were significantly increased compared to the BC group (F=24.527, P<0.05), whereas the expression of caspase-3 mRNA in rats of the HT-12 and HT-24 subgroups was significantly reduced compared to the NT-12 and NT-24 subgroups, respectively (t=6.935, P<0.001; t=4.317, P<0.001). The expression of LC3B-II/I in hippocampus nerve cells of rats in the NT and HT groups significantly increased compared to the BC group (F=6.584, P<0.05), whereas the expression of LC3B-II/I in rats of the HT-12 and HT-24 subgroups significantly reduced compared to the NT-12 and NT-24 subgroups, respectively (t=10.836, P<0.001; t=2.653, P=0.02). Ultrastructure damage of nucleus and mitochondria in the NT group was more evident than in the BC group, and eumorphism of nucleus and mitochondria were maintained in rats of the HT group compared with the NT group. CONCLUSION: Mild hypothermia lessened the injury of nerve cells and improved the neurological function of rats that survived from cardiac arrest by reducing the ROS production of nerve cells and inhibiting the expression of caspase-3 mRNA and LC3, leading to cellular apoptosis and massive autophagy in rats that survived from cardiac arrest after CPR.

Application of enteral nutrition in preoperative bowel preparation for rectal cancer patients undergoing radical operation / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the feasibility and safety of enteral nutrition in preoperative bowel preparation for rectal cancer patients undergoing radical operation.</p><p><b>METHODS</b>Sixty rectal cancer patients undergoing selective low anterior resection were randomized into the trial group(n=30) and the control group(n=30). Patients in the trial group received clean liquid integral protein diet for 3 days before operation without mechanical bowel preparation. Patients in the control group received traditional diet and mechanical bowel preparation. The intraoperative and postoperative clinical data, the quality of bowel preparation, postoperative complications, and nutritional parameters were compared between the two groups.</p><p><b>RESULTS</b>There were no significant differences in clinicopathological characteristics between the two groups before operation. The operative time, blood loss, quality of bowel preparation as well as postoperative hospital stay were not significantly different(all P>0.05). While the time to first flatus [(2.53±0.91) d vs. (3.03±0.68) d] and semi-liquid diet intake[(3.95±0.83) d vs. (4.52±1.14) d] were significantly shorter in the trial group as compared with the control group(all P<0.05). There were no death and no significant difference in postoperative complications [16.7%(5/30) vs. 20.0%(6/30), P>0.05]. The levels of postoperative total protein, albumin, and prealbumin decreased significantly. Meanwhile, the levels of postoperative albumin[(36.2±2.5) g/L vs. (33.5±2.6) g/L, P<0.01] and prealbumin [(325.4±28.2) mg/L vs. (302.5±34.2) mg/L, P<0.01] in the trial group were significantly higher than those in the control group.</p><p><b>CONCLUSIONS</b>Preoperative enteral nutrition can replace the mechanical bowel preparation with better efficacy, and improve the postoperative nutritional status without increasing surgical risk in rectal cancer patients undergoing radical operation.</p>

Effects of Bushen qiangdu decoction on MMP-9 and TIMP-1 produced by peripheral blood mononuclear cells in patients with ankylosing spondylitis / 中国骨伤

<p><b>OBJECTIVE</b>In order to investigate the roles of metalloproteinase in inflammatory bone destruction in ankylosing spondylitis (AS), and analyze the mechanism of preventing inflammatory bone destruction of Bushen Qiangdu decoction (BSQDD) in AS cases. Comparisons were made on the expressions of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) by peripheral blood mononuclear cells (PBMC) between AS patients and healthy controls. The effect of BSQDD was investigated on the expression and of MMP-9 and TIMP-1 produced by PBMC in AS patients.</p><p><b>METHODS</b>From March 2005 to March 2006, 30 active AS cases of Kidney-asthenia, Du-cold and blood-stasis syndrome were selected as patients group in the China-Japan Friendship Hospital. There are 27 male patients and 3 female patients. The age range is from 16 to 45, averaging (30.8 +/- 8.8) years. Disease duration is from 0.5 to 10 years. Cases received three-month BSQDD treatment were considered as the treatment group. Twenty healthy persons were included in the control group. Serum and PBMC were separated. The PBMC were stimulated by PHA and PMA, and the supernatant was collected. The mRNA expression of MMP-9 and TIMP-1 in PBMC was analyzed by RT-PCR. The content of MMP-9 and TIMP-1 in serum and culture supernatant of PBMC were detected by ELISA.</p><p><b>RESULTS</b>Compared with health control group, the serum concentration of MMP-9 and TIMP-1 in patients group before treatment increased (P<0.01, P<0.05), but the level of MMP-9 and TIMP-1 in the serum of patients after treatment decreased compared with pre-treatment cases (P<0.05). Furthermore,compared with health control group, PBMC of patients group before treatment expressed higher levels of MMP-9 and TIMP-1 both on transcript level and at protein level (P<0.01, P<0.05), and the expression levels of MMP-9 and TIMP-1 in PBMC in patients after treatment both on transcript level and at protein level was down-regulated compared with pre-treatment (P<0.01, P<0.05).</p><p><b>CONCLUSION</b>PBMC of AS patients had a higher potential capacity for MMP-9 and TIMP-1. BSQDD possibly prevented inflammatory bone destruction of AS through inhibiting production of MMP-9 and TIMP-1 produced by PBMC.</p>

Morphologic and functional characteristics of the immortalized human liver sinusoidal endothelial cell line / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the morphologic and functional characteristics of the immortalized human liver sinusoidal endothelial cell line (LSEC line).</p><p><b>METHODS</b>Immunofluorescence staining and fluorescence microscopy were used to detect the classic endothelial cell markers in LSEC line, and flow cytometry was used to analyze the purity of the human LSEC line. The morphology (including W-P bodies and surface fenestrations) and phagocytotic capacity of the human LSEC line were observed by transmission and scanning electron microscope. The proliferation curve of the human LSEC line was analyzed by MTT assay. The functional differences between the human LSEC line and human primary LSEC in expression of ELAM-1 and ICAM-1, activities of fibrinolysis (PAI-1, t-PA, u-PA), releasing of IL-6 and IL-8 were compared respectively by enzyme linked immunosorbent assay. Comparison of the susceptibility to hypoxia-reoxygenation induced apoptosis between the human LSEC line and human primary LSEC were investigated by TUNEL.</p><p><b>RESULTS</b>The established human LSEC line maintained a high proliferative ability and has been passaged for more than 80 times in the absence of any growth factors. Immunofluorescence staining showed that the human LSEC line could express classic endothelial cell marks including von Willebrand Factor (vWF), and could take up acetylated low-density lipoproteins (Ac-LDL). The purity of the human LSEC line was confirmed over 95% by flow cytometric analysis. The W-P bodies and the phagocytosis of Dynabeads was demonstrated by transmission electron microscope. And fenestrations could be found cellular surface with scanning electron microscopy. When compared with human primary LSEC, the human LSEC line has an equivalent responsiveness to tumor necrosis factor in up-regulation of ELAM-1 and ICAM-1. The human LSEC line can also release PAI-1, t-PA, u-PA but can not release IL-6 and IL-8 to TNF-alpha. In contrast, human primary LSEC could release IL-6. The human LSEC line showed higher susceptibility to hypoxia-reoxygenation-induced apoptosis, and the percentage of apoptotic cells was as high as (38.4 +/- 6.7)%, while (28.6 +/- 4.5)% and (7.8 +/- 1.2)% respectively in primary LSEC and in human umbilical vein endothelial cells.</p><p><b>CONCLUSIONS</b>The established human LSEC line maintains the special phenotypes and the major functional characteristics, and especially maintains the high susceptibility to hypoxia-reoxygenation-induced apoptosis. Therefore it is feasible to use this cell line for the study of liver ischemia-reperfusion injury.</p>

Study on the status of infection and distribution of rabies virus in China / 中华流行病学杂志

Objective To investigate the status of infection and distribution of rabies virus (RV) in different epidemic areas in China. Methods Brain specimens from animals and suspected patients were collected at the districts of high-, medium- and low incidence rates of human rabies and detected by both direct Immunofluorescence assay (DFA) and RT-PCR. Results 254 of 3007 specimens of dog brains showed RV positive by DFA (positive rate of 8.4% ). Among these 254 samples, 78 showed positive (positive rate of 30.7% ) by RT-PCR. 93 specimens from dogs and cats that had attacked human beings, 63 of them showed positive by DFA (positive rate of 67.7%) and all of them were also positive by RT-PCR. In addition, RV could also be detected in Apodemus agrarius,ferret badger, and suspected patients specimens from the districts under survey. There was no statistical difference between the infection rates of RV in different provinces and regions with different incidence of rabies. Conclusion There might be a relatively high infection rate of RV among the domestic dogs/cats in the endemic areas in China. Wild animals might have been infected with RV in the districts under survey.

Experimental study of the effect of liposome-mediated vascular endothelial growth factor gene on the flap survival in rats / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of VEGF gene on the random flap survival after pedicle division at different time in rats.</p><p><b>METHODS</b>The random-pattern flaps were formed on the back of the 120 SD rats. PcDNAVEGF165 (gene) wrapped with liposome was injected into the flaps in experimental group (n = 40). The flaps in the two control groups were injected with PcDNA (n = 40) or saline (n = 40). 1, 3, 5, 7 days after injection, 10 rats in each group were randomly selected to performed pedicle division. 7 days after pedicle division, the rats were killed to measure the flap survival rate. The microvessels was studied by histologic examination. The expression of VEGF was assessed by immunohistochemical staining. The flaps were also examined under the electron ultrastructure microscopy.</p><p><b>RESULTS</b>1) Flap survival rate after pedicle division in experimental group at 1 day, 3 days, 5 days, 7 days after injection, were (45.45 +/- 12.24)%, (82.95 +/- 3.81)%, (85.00 +/- 3.38)%, (85.96 +/- 3.25)%, respectively. The flap survival rates were significant different between experimental group and the control groups at 3, 5, 7 days after injection (P < 0.05), but not at 1 days after injection (P > 0.05). 2) The average microvascular diameter and number in experimental group were significantly higher than those in control groups (P < 0.05). 3) The expression of VEGF in experimental group was significantly higher than that in the two control groups (P < 0.05). 4) Ultrastructure study showed more angiogenesis in experimental group.</p><p><b>CONCLUSIONS</b>Subcutaneous injection of liposome-mediated VEGF gene can increase the survival rate of flap with early pedicle division. It is a simple, efficient, economic, and the relatively safe gene therapy.</p>

Tumor cell-tumor endothelial cell adhesion mediated by alphavbeta3 and alphavbeta5 molecules / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the role of adhesion molecules alphavbeta3 and alphavbeta5 and their ligands Del-1 and L1 in the tumor-endothelial cell adhesion in vitro.</p><p><b>METHODS</b>The expression of alphavbeta3, alphavbeta5 and ICAM-1 in liver sinusoidal endothelial cells (LSEC) and liver cancer endothelial cells (T3A) cultured under normoxia or hypoxia were analyzed by RT-PCR and fluorescent activated cell sorter (FACS). The expression of Del-1 and L1 in six tumor cell lines under normoxia or hypoxia were analyzed by RT-PCR and Western blot, respectively. The adhesion of dye-labeled tumor cells and endothelial LSEC and T3A cells was measured by a fluorescence plate reader after their culture.</p><p><b>RESULTS</b>The expression of alphavbeta3 and alphavbeta5 were higher in T3A cells than that in LSEC cells, and were upregulated under hypoxia, while the expression of ICAM-1 was lower in T3A cells than that in LSEC cells, and was upregulated under hypoxia only in LSEC. The expression of Del-1 and L1 molecules were obviously different in various tumor cell lines and were differentially regulated under hypoxia. The adhesion of tumor cells with Del-1 or L1 expression was higher in T3A cells than that in LSEC cells, and was significantly increased under hypoxia condition. Furthermore, the adhesion of tumor cells to T3A could be inhibited by antibodies against alphavbeta3 and alphavbeta5, or SiRNAs for beta3 and beta5.</p><p><b>CONCLUSION</b>alphavbeta3 and alphavbeta5 and their ligands Del-1 and L1 may play an important role in tumor cell migration.</p>

Phenotypic and functional characteristics of endothelial cells derived from human liver cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To analyze the phenotypic and functional characteristics of endothelial (T3A) cells derived from human hepatocellular cell carcinoma.</p><p><b>METHODS</b>Endothelial cells were isolated from human hepatocellular carcinoma specimens. The identification of T3A cells was performed by checking von Willebrand Factor (vWF), CD31, CD34 and Dil-Ac-LDL uptake. The cell surface fenestrations, a specific morphological feature of tumor derived EC, were investigated by scanning and transmission electron microscopy. The phenotypic characteristics of T3A cells were analyzed by fluorescence-activated cell sorter (FACS) and were further conformed by real-time PCR at transcription level. Furthermore, tumor necrosis factor alpha (TNFalpha)-induced cytotoxicity was evaluated by 3-(4, 5-dimethythiazolyl) -2, -diphenyl-2H-tetrazolium-bromide (MTT) assay; Matrix metalloproteinase secretion was detected by zymography; Angiogenic ability in vitro was analyzed by culturing T3A cells in three-dimensional Matrigel plug. Coagulant and fibrinolytic activities were detected by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The isolated T3A cells exhibited classic "spindle-shape" morphology and monolayer growth and contact inhibition properties. Immunofluorescent staining showed that T3A cells expressed vWF, CD31, CD34, and uptake of Dil-Ac-LDL at a high level. The cell surface fenestrations were observed on T3A cells by scanning and transmission electron microscopy. By FACS and real-time PCR, T3A cells were found to express alphav3, alphavbeta5 and TNF receptor p75 at high levels, and TNF receptor p55 and ICAM-1 at low levels, as compared with those in human liver sinusoidal endothelial cells (LSEC). In response to TNFalpha, LSEC exhibited a dose-dependent cytotoxicity, while T3A cells were resistant. Gelatin zymography showed that MMP-2 activity was higher in T3A cells than that in LSEC. In a three-dimensional plug of Matrigel, T3A cells exhibited stronger angiogenic ability as compared with LSEC. In addition, T3A cells released more tissue factor (TF), tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor (PAI-1) and urine plasminogen activator (u-PA) than LSEC in response to TNFalpha.</p><p><b>CONCLUSION</b>Tumor-derived endothelial cells are phenotypically and functionally different from those derived from normal liver tissue.</p>

The early diagnosis and therapy of aneurismal subarachnoid hemorrhage / 中华外科杂志

<p><b>OBJECTIVE</b>To discuss the early diagnostic methods and therapeutic principles of aneurysmal subarachnoid hemorrhage (SAH), and evaluate the therapeutic efficacy objectively.</p><p><b>METHODS</b>Using neuro-imaging examinations combined with case history and clinical symptoms to make the early diagnosis of 96 case with aneurysmal SAH, and Guglielmi detachable microcoil (GDC) was utilized for early intracapsular embolization in the ruptured aneurysms. Efficient symptomatic treatment was done early after operation.</p><p><b>RESULTS</b>All of 96 cases were early diagnosed and successfully embolized; Among them, the aneurysmal lumen was 100% occluded in 83 cases, 95% in 8 cases, 90% in 5 cases. There were 3 cases complicating with aneurysms rupture during operation, 5 cases with cerebral vasospasm. One case was affected by microcoil terminal escape after operation, 3 recurrent cases were all cured with secondary GDC embolization. There were 9 complications associated with embolization techniques and 13 cases (13.5%) occurring permanent sequelae associated with SAH. According to the Glasgow prognosis score, 77 patients got grade I, 7 grade II, 6 grade III, 3 grade IV, and 3 grade V. The mortality rate was 3.1%.</p><p><b>CONCLUSIONS</b>To make early etiological diagnosis of the SAH patients, using GDC to embolize the aneurysms, and earlier efficient symptomatic treatment are important methods to improve the curative rate and reduce the mortality rate.</p>

Exploration for building PACS and RIS / 中国医疗器械杂志

This article elaborates key points of the PACS and RIS project: its overall planning, implementation step by step, integration of PACS and HIS based on IHE, and the prudent selection of partners and so on.