Associação da hipertensão com a gravidade e a mortalidade de pacientes hospitalizados com COVID-19 em Wuhan, China: Estudo unicêntrico e retrospectivo / Association of hypertension with severity and mortality in hospitalized patients with COVID-19 in wuhan, China: A single-centered, retrospective study

Resumo Fundamento A doença Coronavírus 2019 (COVID-19), causada pela síndrome respiratória aguda grave Coronavírus 2 (SARS-CoV-2), espalhou-se pelo mundo. Objetivo Investigar a associação entre a hipertensão e a gravidade/mortalidade de pacientes hospitalizados com COVID-19 em Wuhan, China. Métodos Um total de 337 pacientes diagnosticados com COVID-19 no Sétimo Hospital da cidade de Wuhan, de 20 de janeiro a 25 de fevereiro de 2020, foram inseridos e analisados em um estudo de caso unicêntrico e retrospectivo. O nível de significância adotado para a análise estatística foi 0,05. Resultados Dos 337 pacientes com diagnóstico confirmado de COVID-19, 297 (87.8%) tiveram alta do hospital e 40 pacientes (22,9%) morreram. A idade média foi de 58 anos (variando de 18 a 91 anos). Havia 112 (33,2%) pacientes diagnosticados com hipertensão no momento da internação (idade média, 65,0 anos [variação, 38-91 anos]; sendo 67 homens [59,8%, IC95%: 50,6%-69,0%], p=0,0209). Pacientes com hipertensão apresentaram uma porção significativamente maior de casos graves (69 [61,6%, IC95%: 52,5%-70,8%] vs. 117 [52,0%, IC95%: 45,4%-58,6%] em pacientes graves e 23 [19,3%, IC95%: 12,9%-28,1%] vs. 27 [12,0%, IC95%: 7,7%-16,3%] em pacientes críticos, p=0,0014) e maiores taxas de mortalidade (20 [17,9%, IC95%: 10,7%-25,1%] vs. 20 [8,9%, IC95%: 5,1%-12,6%, p=0,0202). Além disso, pacientes hipertensos apresentaram níveis anormais de vários indicadores, como linfopenia e inflamação, e nas funções cardíacas, hepáticas, renais e pulmonares no momento da internação. O grupo de pacientes com hipertensão também demonstrou níveis maiores de TNT e creatinina próximo da alta. Conclusão A hipertensão está altamente associada à gravidade ou mortalidade da COVID-19. Um tratamento agressivo deve ser considerado para pacientes hipertensos com COVID-19, principalmente com relação a lesões cardíacas e dos rins.

Abstract Background Coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has spread worldwide. Objective To investigate the association between hypertension and severity/mortality in hospitalized patients with COVID-19 in Wuhan, China. Methods A total of 337 patients diagnosed with COVID-19 at the Seventh Hospital of Wuhan City, from January 20 to February 25, 2020, were enrolled and analyzed in a retrospective, single-center case study. The significance level adopted in the statistical analysis was 0.05. Results Of the 337 patients with confirmed diagnosis of COVID-19, 297 (87.8%) were discharged from the hospital and 40 patients (22.9%) died. The median age was 58 years (range, 18-91 years). There were 112 (33.2%) patients diagnosed with hypertension at admission (median age, 65.0 years [range, 38-91 years]; 67 [59.8%, 95%CI: 50.6%-69.0%] men, p=0.0209). Patients with hypertension presented a significantly higher portion of severe cases (69 [61.6%, 95%CI:52.5%-70.8%] vs. 117 [52.0%, 95%CI: 45.4%-58.6%] in severe patients and 23 [19.3%, 95%CI:12.9%-28.1%] vs. 27 [12.0%, 95%CI: 7.7%-16.3%] in critical patients, p=0.0014) and higher mortality rates (20 [17.9%, 95%CI: 10.7%-25.1%] vs. 20 [8.9%, 95%CI: 5.1%-12.6%, p=0.0202). Moreover, hypertensive patients presented abnormal levels of multiple indicators, such as lymphopenia, inflammation, heart, liver, kidney, and lung function at admission. The hypertension group still displayed higher levels of TnT and creatinine at approaching discharge. Conclusion Hypertension is strongly associated with severity or mortality of COVID-19. Aggressive treatment may be considered for COVID-19 patients with hypertension, especially regarding cardiac and kidney injury.

Expression of hypoxia-inducible factor-1 in mice infected with Toxoplasma gondii during pregnancy / 中国血吸虫病防治杂志

Objective To investigate the expression and possible role of hypoxia-inducible factor-1 (HIF-1) at the maternal-fetal interface following Toxoplasma gondii infection during early pregnancy. Methods Twenty pregnant C57BL/6 mice, each weighing 16 to 20 g, were randomly divided into 4 groups, including the 12-d control group, 12-d infection group, 18-d control group and 18-d infection group. Mice in the 12-d and 18-d infection groups were injected intraperitoneally with 150 tachyzoites of the T. gondii PRU strain on day 6 of pregnancy, while mice in the 12-d control and 18-d control groups were injected with the same volume of phosphate buffered saline (PBS). Mice in the control and infection groups were sacrificed on days 12 and 18 of pregnancy, and the placental and uterine specimens of the pregnant mice in each group were sampled for pathological examinations. The mRNA expression of HIF-1α, HIF-1β and vascular endothelial growth factor (VEGF) was quantified using quantitative fluorescent real-time PCR (qPCR) assay in the placental and uterine specimens, and the correlation between HIF-1α and VEGF mRNA expression was examined. In addition, and the HIF-1α expression was detected using immunohistochemical staining in the placental and uterine specimens of pregnant mice. Results Compared with the 12-d and 18-d control groups, adverse pregnant outcomes were observed in mice in 12-d and 18-d infection groups, such as teratism and placental dysplasia. HE staining showed swelling and blood stasis of cells, sinusoid reduction and inflammatory cell infiltration in the labyrinth area of the placenta specimens of mice in 12-d and 18-d infection groups relative to 12-d and 18-d control groups, and columnar epithelial cell injury and inflammatory cell infiltration were seen in the mouse uterine specimens in both infection groups. qPCR assay detected significantly higher HIF-1α (F = 132.6, P < 0.05) and HIF-1β mRNA expression (F = 286.9, P < 0.05) in the placental specimens and lower HIF-1α (F = 111.5, P < 0.05) and HIF-1β mRNA expression (F = 55.2, P < 0.05) in the uterine specimens in the 12-d infection group than in the 12-day control group, and significantly lower HIF-1α and HIF-1β mRNA expression was detected in the placental and uterine specimens in the 18-d infection group than in the 18-day control group (F = 215.8, 418.9, 156.8 and 200.1; all P values < 0.05). Significantly lower VEGF-A (F = 426.2, P < 0.05), VEGF-B (F = 104.6, P < 0.05) and VEGF-C mRNA expression (F = 566.9, P < 0.05) in the placental specimens and higher VEGF-A (F = 426.2, P < 0.05), VEGF-B (F = 104.6, P < 0.05) and VEGF-C mRNA expression (F = 566.9, P < 0.05) in the uterine specimens were detected in the 12-d infection group than in the 12-d control group, and higher VEGF-A, VEGF-B and VEGF-C mRNA expression was found in the placental and uterine specimens in the 18-d infection group than in the 18-d control group (F = 521.9, 100.6, 275.9, 224.6, 108.2 and 333.4; all P values < 0.05). Immunohistochemical staining showed strongly and mildly positive HIF-1α expression in the mouse placental labyrinth area in the 12-d and 18-d infection groups relative to 12-d and 18-d control groups, while no HIF-1α expression was detected in mouse uterine specimens. Conclusions HIF-1α expression appears a tendency towards a rise in the second trimester and a reduction in the third trimester in mice following T. gondii infection during early pregnancy, which is contrary to the changing tendency of VEGF-A, VEGF-B, and VEGF-C expression. It is hypothesized that HIF-1α inhibits placental angiogenesis in mice during pregnancy through suppressing VEGF expression, resulting in adverse pregnant outcomes.

PD-1/PD-L1 expression and its interaction with interferon-γ in Toxoplasma gondii-infected mice at middle and late pregnancy / 中国血吸虫病防治杂志

Objective To explore the dynamic expression of programmed cell death-1 (PD-1) and its ligand PD-L1 at the maternal-fetal interface of mice post-infection with Toxoplasma gondii at early pregnancy and examine its interaction with interferon-γ (IFN-γ). Methods A total of 20 mice at day 0 of pregnancy were randomly assigned into 4 groups, including the 12-day pregnancy control group (12 dpn group), 12-day pregnancy and infection group (12 dpi group), 18-day pregnancy control group (18 dpn group) and 18-day pregnancy and infection group (18 dpi group), respectively. On the 6th day of the pregnancy, mice in the 12 dpi and 18 dpi groups were injected intraperitoneally with 150 tachyzoites of the T. gondii PRU strain, while mice in the 12 dpn and 18 dpn groups were injected with the same volume of PBS. All mice in the four groups were sacrificed on 12th and 18th day of the pregnancy, and the number of placenta and fetus was counted and the weight of placenta and fetus was measured. Then, the placental and uterine tissues of the pregnant mice in each group were sampled for pathological examinations. The mRNA expression of PD-1, PD-L1, T. gondii surface antigen SAG-1 and IFN-γ genes was quantified using a quantitative real-time PCR (qPCR) assay, and the correlation between PD-1 and IFN-γ expression was examined. In addition, the 12 dpn group, 12 dpi group, 18 dpn group, 18 dpi group, PBS negative control of the 12 pdi group and PBS negative control of the 18 dpi group were assigned, and the PD-1 expression was determined in the uterine and placenta tissues of the pregnant mice. Results Adverse pregnant outcomes were seen in mice in the 12 dpi and 18 dpi groups, including placental dysplasia and fetal maldevelopment, and the placental weights and fetal body weights were significantly lower in mice in the 12 dpi and 18 dpi groups than those in the 12 dpn and 18 dpn groups (t = 5.52, 11.44, 12.63 and 11.67, all P < 0.01). The histopathological examinations showed that the decidua and junctional regions of the placental tissues were loosely connected in the 12 dpi and 18 dpi groups, and a large number of inflammatory cells infiltration and congestion were seen in the placental and uterine tissues. qPCR assay detected significant differences in PD-1, PD-L1, IFN-γ and SAG-1 expression in the placental and uterine tissues among the 12 dpn, 12 dpi, 18 dpn and 18 dpi groups (F = 22.48, 51.23, 9.61, 47.49, 16.08, 21.52, 28.66 and 238.90, all P < 0.05), and the PD-1, PD - L1, IFN - γ and SAG - 1 expression was all significantly higher in the placental and uterine tissues of mice in the 12 dpi group than in the 12 dpn group (all P values < 0.05). The PD-1 and PD-L1 expression was significantly lower in the placental tissues of mice in the 18 dpi group than in the 18 dpn group (all P values < 0.05), and the IFN-γ and SAG-1 expression was significantly higher in the placental and uterine tissues of mice in the 18 dpi group than in the 18 dpn group (all P values < 0.05), while the PD-1 and PD-L1 expression was significantly lower in the placental and uterine tissues of mice in the 18 dpi group than in the 12 dpi group (all P values < 0.05). Immunohistochemical staining showed PD-1 expression in the inflammatory cells of the placental tissues of mice in the 12 dpi group, and no apparent PD-1 expression in the 18 dpi group, while strongly positive PD-1 expression was found in the uterine epithelium of mice in the 12 dpi group, and mildly strong expression was in the 18 dpi group. In addition, the IFN-γ mRNA expression was positively correlated with the PD-1 mRNA expression in placental (rs = 0.99, P < 0.01) and uterine tissues of mice in the 12 dpi group (rs = 0.97, P < 0.01) and in placental (rs = 0.82, P < 0.01) and uterine tissues of mice in the 18 dpi group (rs = 0.81, P < 0.01). Conclusions Following T. gondii infection at early pregnancy, the PD-1 and PD-L1 expression shows a remarkable rise at middle pregnancy and a reduction at late pregnancy in placental and uterine tissues of mice, which appears the same tendency with IFN-γ expression during the same time period, and PD-1 expression positively correlates with IFN-γ expression. The dynamic expression of PD-1 and PD-L1 on the maternal-fetal interface of mice may be mutually mediated by IFN-γ induced by T. gondii infection.

Study on the mechanisms of the intestinal tight-junction destruction caused by Blastocystis hominis infection in rats / 中国血吸虫病防治杂志

Objective To explore the mechanism of the intestinal barrier damage caused by Blastocystis hominis infections in rats. Methods Thirty SD rats were randomly divided into the control group, and the 1-, 3-, 6- and 9-week-infection groups, of 6 rats in each group. Rats in each infection group were orally infected with B. hominis trophozoites at a density of 2 × 108 parasites per rat, and the control group was given an equal volume of phosphate buffered saline solution. The 7-hour urine samples were collected 1, 3, 6 and 9 weeks post-infection for the measurement of the intestinal permeability. Then, rats were sacrificed using the cervical dislocation method, and the cecum specimens were collected for the detection of the intestinal epithelial cell permeability. The expression of tight junction-related Occludin and Claudin - 1 genes and apoptosis-related Bcl - 2 and Bax genes was quantified in cecum epithelial cells using the real-time fluorescent quantitative PCR (qPCR) assay, and cell apoptosis was detected in the rat cecum using the TdT-mediated dUTP nick-end labeling (TUNEL) assay. Results The median urinary lactolose to mannitol ratios were 0.29, 0.72, 0.44, 0.46 and 0.38 in the control group, and the 1-, 3-, 6- and 9-week-infection groups, respectively, and the difference was statistically significant (H = 12.09, P < 0.05). B. hominis invasion and epithelial injury were observed in intestinal epithelial cells of rats infected with B. hominis, and transmission electron microscopy displayed the destruction of tight junctions between intestinal epithelial cells. The relative expression of Occludin, Claudin-1, Bcl-2 and Bax genes was 1.04, 0.62, 0.71, 0.68 and 0.96; 1.03, 0.61, 0.63, 0.76 and 0.86; 1.08, 0.70, 0.75, 0.74 and 1.03; and 1.00, 1.57, 1.33, 1.35 and 1.10 in the control group and the 1-, 3-, 6- and 9-week-infection groups, respectively, and all differences were statistically significant (F = 2.86, 2.85, 3.37 and 4.45, all P values < 0.05). The median number of positive staining cells were 1.00, 13.00, 9.00, 3.50 and 1.00 in rat cecum specimens in the control group, and the 1-, 3-, 6- and 9-week-infection groups, respectively, and the difference was statistically significant (H = 22.95, P < 0.01). Conclusion B. hominis infection may cause an increase in the rat intestinal permeability through triggering the apoptosis of intestinal epithelial cells to destroy the tight junction between intestinal epithelial cells, thereby destroying the intestinal barrier function.

Novel coronavirus pneumonia (COVID-19) after pulmonary surgery: A case report / 中国胸心血管外科临床杂志

@#There was a male novel coronavirus (2019-nCoV, SARS-CoV-2) pneumonia (COVID-19) patient after pulmonary surgery at age of 61 years. The patient had no clear history of contact COVID-19 patient before surgery. He developed transient fever on the 4th day after surgery. The body temperature returned to normal on the 5th day after antibiotic adjustment. The patient developed fever and fatigue again on the 6th day after surgery. A chest CT scan revealed postoperative pneumonia. The patient was treated by ganciclovir and moxifloxacin hydrochloride. The patient's temperature gradually decreased on the 7th to 9th days after the operation. CT scan on the 10th day after surgery showed viral pneumonia, so we immediately raised the level of protection. The novel coronavirus nucleic acid test was positive. The patient was immediately transferred to the designated hospital for treatment. The patient was treated by arbidol, moxifloxacin, human immunoglobulin (PH4), ambroxol and other nutritional symptomatic and supportive treatment. The patient's condition is currently stable. Ten people in close contact with the patient developed symptoms, and their CT scans showed viral pneumonia. Six of them were positive in nucleic acid tests, and the others were still under quarantine observation. This shows that it is easy to confuse the imaging manifestations of pneumonia with novel coronavirus pneumonia after lung surgery. We should perform nucleic acid detection as soon as possible in the early diagnosis of CT and reformulate the treatment protocol.

Comparative study on the application of non-intubated anesthesia in thoracoscopic lobectomy / 中国胸心血管外科临床杂志

@#Objective To analyze the feasibility and advantages of non-intubated anesthesia in thoracoscopic lobectomy. Methods The clinical data of 59 patients with thoracoscopic lobectomy and non-intubated anesthesia in the Department of Thoracic Surgery, Tongji Hospital from January 2015 to December 2017 were retrospectively reviewed, including 24 males and 35 females, aged 56.86±7.13 years (an observation group); 59 patients with thoracoscopic lobectomy undergoing general anesthesia with tracheal intubation in the same period were randomly selected, as a control group, including 27 males and 32 females, aged 55.37±6.86 years. Complications such as airway injury, refractory cough, pharyngalgia, nausea and vomiting were compared between the two groups. Postoperative inflammatory factor levels, postoperative hospital stay, and intraoperative and postoperative hospitalization costs were also compared. Results There was no difference between the two groups in general conditions such as age, gender, body mass index. There was also no difference in operation time, intraoperative bleeding volume or lymph node dissection. But the observation group had lower levels of procalcitonin and C reactive protein at postoperative 1 d (0.12±0.51 ng/ml vs. 0.14±0.70 ng/ml, P=0.03; 11.30±3.60 mg/L vs. 13.33±4.41 mg/L, P=0.01), lower rate of postoperative complications of refractory cough, pharyngalgia, nausea and vomiting (3.38% vs. 15.25%, P=0.03; 5.08% vs. 20.33%, P=0.01; 3.38% vs. 15.25%, P=0.03), less retain time of thoracic duct, postoperative hospital stay, and lower intraoperative and postoperative hospitalization costs (5.89±1.37 d vs. 7.00±1.73 d, P=0.00; 10.01±1.85 d vs. 11.37±2.45 d, P=0.00; 53 810.94±5 745.44 yuan vs. 58 223.16±6 445.08 yuan, P=0.00). Conclusion Thoracoscopic lobectomy with non-intubated anesthesia can avoid traditional airway injury caused by endotracheal intubation, reduce postoperative symptoms such as refractory cough, pharyngalgia, nausea and vomiting caused by general anesthesia, reduce or even avoid lung injury caused by one-side lung ventilation, promote recovery after surgery, reduce antibiotic use, and shorten hospital stay, which is more consistent with the requirements of the concept of overall minimal invasiveness and enhanced recovery.

Effect of cordycepin on apoptosis and autophagy of tongue cancer cells in vitro and the molecular mechanism / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of cordycepin on cell cycle, apoptosis and autophagy of human tongue cancer TCA-8113 cells and explore the mechanism of cordycepin for inhibiting the occurrence of tongue cancer.</p><p><b>METHODS</b>CCK-8 method was used to assess the inhibitory effect of cordycepin on TCA-8113 cell proliferation in vitro. The cell cycle and cell apoptosis of TCA-8113 cells treated with different concentrations of cordycepin were analyzed using flow cytometry. The expressions of apoptosis-related genes caspase-3, caspase-9, Bcl-2, and Bax were examined using quantitative real-time PCR and Western blotting, and immunohistochemistry was used to detect the expressions of autophagy-related proteins LC-3β, P62, p-mTOR, and AMPK.</p><p><b>RESULTS</b>CCK-8 assay showed that cordycepin significantly inhibited the proliferation of TCA-8113 cells in a concentration-dependent manner with an IC of 3.548 mg/mL at 24 h and an IC of 1.185 mg/mL at 48 h. Flow cytometric analysis showed that cordycepin caused cell cycle arrest at S phase and dose-dependently increased the apoptotic rate of TCA-8113 cells. Treatment of the cells with cordycepin enhanced the expressions of Bax, caspase-3 and caspase-9 at both the mRNA and protein levels and inhibited the expression of the antiapoptotic gene Bcl-2. Immunohistochemistry demonstrated that cordycepin promoted the expression of LC-3β and AMPK and inhibited the expression of P62 and p-mTOR.</p><p><b>CONCLUSION</b>Cordycepin inhibits the proliferation and induces apoptosis of HCT-116 cells through the mitochondrial pathway and induces autophagy via the AMPK/mTOR pathway.</p>

Changes in C-reactive protein and procalcitonin levels in neonates with necrotizing enterocolitis and their clinical significance / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the changes in C-reactive protein (CRP) and procalcitonin (PCT) levels in neonates with necrotizing enterocolitis (NEC) and their clinical significance.</p><p><b>METHODS</b>According to the modified Bell's staging criteria, 142 neonates with NEC were divided into stage I group (n=40), stage II group (n=72), and stage III group (n=30). All the 18 neonates who underwent surgical treatment had stage III NEC, and among the 124 neonates who underwent conservative treatment, 12 had stage III NEC and the others had stage I or II NEC. CRP and PCT were measured before treatment, on the next day after treatment, and during the recovery stage.</p><p><b>RESULTS</b>Before treatment, on the next day after treatment, and during the recovery stage, the stage III group had a higher level of CRP than the stage I and stage II groups (P<0.05). On the next day after treatment, the stage II and stage III groups had an increase in CRP (P<0.05), and the stage III group had an increase in PCT (P<0.05). The stage II and stage III groups had lower CRP and PCT in the recovery stage than before treatment and on the next day after treatment (P<0.05). The stage III group had higher incidence rate of respiratory failure and rate of mechanical ventilation than the stage I and stage II groups (P<0.05), and the stage III group had a higher incidence rate of sepsis than the stage II group (P=0.010). Gastrointestinal perforation and intestinal stenosis were observed in 10 and 8 neonates respectively in the stage III group. CRP on the next day after treatment had a value in predicting stage III NEC (P<0.05), and CRP before treatment and on the next day after treatment had a value in predicting the need for surgery (P<0.05).</p><p><b>CONCLUSIONS</b>Levels of CRP and PCT and their changes can help with the early diagnosis of Bell stage II/III NEC, and CRP can be used to predict the development of stage III NEC and the need for surgery.</p>

Down-regulation of p110β Expression Increases Chemosensitivity of Colon Cancer Cell Lines to Oxaliplatin / 华中科技大学学报（医学）（英德文版）

This study examined the synergetic effect of class IA Phosphoinositide 3-kinases catalytic subunit p110β knockdown in conjunction with oxaliplatin treatment on colon cancer cells.Down-regulation of p110β by siRA interference and oxaliplatin treatment were applied in colon cancer cell lines HT29,SW620 and HCT116.MTT assay was used to measure the inhibitory effect of p110 knockdown on the proliferation of colon cancer cell lines.SubG1 assay and Annexin-V FITC/PI double-labeling cytometry were applied to detect cell apoptosis.And cell cycle was evaluated by using PI staining and flow cytometry.The expression of caspase 3,cleaved PARP,p-Akt,T-Akt and p 110β was dctermined by western blotting.The results suggested that down-regulation of p110β expression by siRNA obviously reduced cell number via accumulation in G0-G1 phase of the cell cycle in the absence of notablely increased apoptosis in colon cancer cell lines HT29 and SW620 (S phase arrest in -HCT116).Moreover,inhibition of p110β expression increased oxaliplatin-induced cell apoptosis and cell cycle arrest in HT29,HCT116 and SW620 cell lines.In addition,increases of cleaved caspase-3 and cleaved PARP induced by oxaliplatin treatment were determined by immunoblotting in p110β knockdown group compared with normal control group and wild-type group.It is concluded that down-regulated expression of p110β could inhibit colon cancer cells proliferation and result in increased chemosensitivity of colorectal cancer cells to oxaliplatin through augmentation of oxaliplatin-induced cell apoptosis and cell cycle arrest.

Comparison of transurethral detaching resection and plasmakinetic transurethral resection of the prostate for benign prostatic hyperplasia / 中华男科学杂志

<p><b>OBJECTIVE</b>To compare the clinical effects of transurethral detaching resection (TUDRP) and plasmakinetic transurethral resection of the prostate (PKRP) for benign prostatic hyperplasia (BPH).</p><p><b>METHODS</b>We collected by the single blind method the clinical data of 81 cases of BPH treated by PKRP (n=41) and TUDRP (n=40), and compared the two groups of patients in their age, preoperative prostate volume, weight of the resected gland, operation time, intraoperative bleeding, time of postoperative Foley's catheter retention, and pre- and post-operative IPSS.</p><p><b>RESULTS</b>The post-operative IPSS was significantly higher in the PKRP than in the TUDRP group (9.95 +/- 1.54 vs. 8.70 +/- 1.13, t = 0.0029, P = 0.0059).</p><p><b>CONCLUSION</b>TUDPR has a better clinical effect than PKRP in the treatment of BPH.</p>

High mobility group box-1 stimulates proinflammatory cytokine production in endothelial cells via MAP kinases / 南方医科大学学报

<p><b>OBJECTIVE</b>To examine the synergistic effect of recombinant human high mobility group box 1 (HMGB1) protein and lipopolysaccharides (LPS) on the release of interleukin-8 (IL-8) and monocyte chemotactic protein 1 (MCP-1) in human umbilic vein endothelial cells (HUVECs), and explore the role of mitogen-activated protein kinases (MAPK) signal transduction in cytokine release.</p><p><b>METHODS</b>HUVECs were incubated with recombinant HMGB1 (0-75 ng/ml) for 24 h and the culture medium supernatant was harvested for detection of IL-8 and MCP-1 with LiquiChip system. At 0, 1, 3, 6, 12 and 24 h after stimulation with 15 ng/ml HMGB1 or 15 ng/ml HMGB1 plus 10 ng/ml LPS, the levels of IL-8 and MCP-1 in the HUVECs were examined. To test the effect of MAPK inhibitors, HUVCs were pretreated with the inhibitors SB203580 (20 mol/L), PD98059 (20 mol/L), and JNK inhibitor II (50 nmol/L) 1 h before HMGB1 and LPS stimulation.</p><p><b>RESULTS</b>The levels of IL-8 and MCP-1 were significantly increased in the HUVECs stimulated with HMGB1 protein at the concentrations of 3, 15 and 75 ng/ml in comparison with the control levels (P<0.01). Since 3-6 h after the stimulation with HMGB1, the levels of IL-8 and MCP-1 began to increase gradually, and steadily increased at 12 and 24 h, all significantly higher than those of the control group (P<0.01). Stimulation of the HUVECs with LPS (10 ng<ml) or HMGB1 (15 ng/ml) alone resulted in significantly increased levels of IL-8 and MCP-1 (P<0.01), which were further increased after costimulation with LPS and HMGB1, suggesting a synergistic effect between HMGB1 and LPS (P<0.01). This synergistic effect was significantly inhibited by pretreatment with MAPK signaling kinases inhibitors, especially the p38 MAP kinase inhibitor SB203580, and the cocktail of MAP kinase inhibitors almost totally blocked the expression of these chemokines in HUVECs treated with HMGB1 and LPS.</p><p><b>CONCLUSION</b>HMGB1 protein can activate HUVECs to produce the chemokines IL-8 and MCP-1 in a dose- and time-dependent manner. HMGB1 also acts synergistically with LPS to induce IL-8 and MCP-1 release, which might play an important role in the development of sepsis. MAPK signal transduction plays an important role in HMGB1 and LPS-induced IL-8 and MCP-1 release.</p>

Construction of a eukaryotic expression vector for alpha-1-antitrypsin and the localization of the expression product in NIH 3T3 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a eukaryotic expression vector for alpha-1-antitrypsin (AAT) and detect its expression and localization in NIH 3T3 cells.</p><p><b>METHODS</b>The total RNA was extracted from the liver tissue of BALB/c mice, and the corresponding coding sequences for mouse AAT (GenBank accession No. NM_009243) were amplified by RT-PCR and cloned into hemagglutinin (HA)-tagged vector pcDNA3-HA. The construct was then transfected into NIH 3T3 cells, which were observed under fluorescence microscope.</p><p><b>RESULTS</b>The recombinant plasmid was verified by PCR, enzyme digestion and sequence analysis, and the fusion protein was highly expressed in NIH 3T3 cells. Under fluorescence microscope, the fusion protein was found to distribute mainly in the cytoplasm.</p><p><b>CONCLUSION</b>The expression vector for AAT-HA fusion protein has been successfully constructed and effectively expressed in mammalian cells to allow future functional study of AAT in mammalian cells.</p>

Construction of Recombinant Adenovirus Carrying GRIM19 and Its Effect on SW480 Cells / 华中科技大学学报（医学）（英德文版）

In order to examine the effect of GRIM 19 on colon cancer cell SW480, the recombinant adenovirus carrying GRIM19 gene was constructed and transfected into SW480 cells. GRIMI9 cDNA was amplified by PCR with the template pcxn2-GRlMl9 and cloned into the shuttle plasmid pAdTrack-CMV. The plasmid pAdTrack-CMV-GRIM19 was linearized by PmeI and homologously recombined with bone plasmid pAdEasy-1 in BJ5183, followed by identification by enzyme diges- tion. After transfection of linearized pAd-GRIM19 with PacI into HEK293 cells, Ad-GRIMI9 was obtained and amplified by 3 circles. SW480 cells were infected with Ad-GRIM19. The apoptosis rate was detected by flow cytometry. Agarose electrophoresis revealed the bands of recombinant plasmids identified by enzyme digestion were in the right range corresponding with expectation. Under the fluorescent microscopy, the package of Ad-GRIM19 in HEK293 cells and the expression of Ad-GRIM19 in SW480 cells were observed. The transfection of Ad-GRIM19 into SW480 cells in-creased the apoptosis rate of SW480 cells as compared with controls. It was concluded that Ad-GRIM19 was successfully constructed and the overexpression of GRIM19 in colon cancer cell lines could promote the apoptotic cell death.

Effects of acupuncture plus acupoint sticking on bone mass density and pain in the patient of primary osteoporosis / 中国针灸

<p><b>OBJECTIVE</b>To observe effects of acupuncture plus acupoint sticking of Migudan on bone mass density (BMD) and pain in the patient of primary osteoporosis.</p><p><b>METHODS</b>By computer random number generating method, the patients were randomly divided into a treatment group treated with acupuncture plus acupoint sticking of Migudan, and a control group treated with Gaitianli, 48 cases in each group. Changes of their BMD and cumulative scores of pain after treatment were investigated.</p><p><b>RESULTS</b>Acupuncture plus acupoint sticking of Migudan could relieve cumulative score of bone pain in the patient of primary osteoporosis as compared with the control group (P 0.01), with 95% CI= 4.05 to approximately 4. 31 points; acupuncture plus acupoint sticking of Migudan could increase significantly BMD in the patients as compared with the control group (P < 0.05), L2 to approximately L4 BMD 95% CI = 0.029 to approximately 0.073 g/cm2, the neck of femur BMD 95% CI = 0.013 to approximately 0.047 g/cm2.</p><p><b>CONCLUSION</b>Acupuncture plus acupoint sticking of Migudan has definite therapeutic effect on primary osteoporosis.</p>

Effect of microbial elicitors on sclerotia biomass and carotenoid yield in surface cultures of Penicillium sp.PT95 / 生物工程学报

Six kinds of elicitors were prepared respectively from Neurospora crassa, Monascus purpureus, Sporobolomyces roseus, Rhodotorula rubra, Nocardia sp. N89 and Actinoplanes sp. A05. When Penicillium sp. PT95 was incubated in Czapek's agar plates containing appropriate amounts of elicitors, both its sclerotia biomass and carotenoid content accumulated in sclerotia were enhanced significantly (P < 0.01). Among tested elicitors, the elicitors from the fungi N. crassa, M. purpureus, S.-roseus and R. rubra were more effective than those from the actinomycetes Nocardia sp. N89 and Actinoplanes sp. A05; the elicitor from M. purpureus gave the highest carotenoid yield of 599 micrograms/plate, 2.76 times higher than that of control. Every one of elicitors except that from M. purpureus could increase significantly the proportion of beta-carotene in total carotenoids (P < 0.01).