Antifungal Activity and Action Mechanism Study of Coumarins from Cnidium monnieri Fruit and Structurally Related Compounds.

The increasing resistance of plant diseases caused by phytopathogenic fungi highlights the need for highly effective and environmentally benign agents. The antifungal activities of Cnidium monnieri fruit extracts and five isolated compounds as well as structurally related coumarins against five plant pathogenic fungi were evaluated. The acetone extract, which contained the highest amount of five coumarins, showed strongest antifungal activity. Among the coumarin compounds, we found that 4-methoxycoumarin exhibited stronger and broader antifungal activity against five phytopathogenic fungi, and was more potent than osthol. Especially, it could significantly inhibit the growth of Rhizoctonia solani mycelium with an EC50 value of 21 µg mL-1 . Further studies showed that 4-methoxycoumarin affected the structure and function of peroxisomes, inhibited the ß-oxidation of fatty acids, decreased the production of ATP and acetyl coenzyme A, and then accumulated ROS by damaging MMP and the mitochondrial function to cause the cell death of R. solani mycelia. 4-Methoxycoumarin presented antifungal efficacy in a concentration- dependent manner in vivo and could be used to prevent the potato black scurf. This study laid the foundation for the future development of 4-methoxycournamin as an alternative and friendly biofungicide.

A multicenter retrospective study on survival rate and complications of very preterm infants. / æžæ—©äº§å„¿å­˜æ´»çŽ‡å’Œå¹¶å‘ç—‡çš„å¤šä¸­å¿ƒå›žé¡¾æ€§ç ”ç©¶.

OBJECTIVES: To study the survival rate and the incidence of complications of very preterm infants and the factors influencing the survival rate and the incidence of complications. METHODS: The medical data of the very preterm infants with a gestational age of <32 weeks and who were admitted to the Department of Neonatology in 11 hospitals of Jiangsu Province in China from January 2018 to December 2019 were retrospectively reviewed. Their survival rate and the incidence of serious complications were analyzed. A multivariate logistic regression analysis was used to evaluate the risk factors for death and serious complications in very preterm infants. RESULTS: A total of 2 339 very preterm infants were enrolled, among whom 2 010 (85.93%) survived and 1 507 (64.43%) survived without serious complications. The groups with a gestational age of 22-25+6 weeks, 26-26+6 weeks, 27-27+6 weeks, 28-28+6 weeks, 29-29+6 weeks, 30-30+6 weeks, and 31-31+6 weeks had a survival rate of 32.5%, 60.6%, 68.0%, 82.9%, 90.1%, 92.3%, and 94.8% respectively. The survival rate tended to increase with the gestational age (P<0.05) and the survival rate without serious complications in each gestational age group was 7.5%, 18.1%, 34.5%, 52.2%, 66.7%, 75.7%, and 81.8% respectively, suggesting that the survival rate without serious complications increased with the gestational age (P<0.05). The multivariate logistic regression analysis showed that high gestational age, high birth weight, and prenatal use of glucocorticoids were protective factors against death in very preterm infants (P<0.05), and 1-minute Apgar score ≤3 was a risk factor for death in very preterm infants (P<0.05); high gestational age and high birth weight were protective factors against serious complications in very preterm infants who survived (P<0.05), while 5-minute Apgar score ≤3 and maternal chorioamnionitis were risk factors for serious complications in very preterm infants who survived (P<0.05). CONCLUSIONS: The survival rate is closely associated with gestational age in very preterm infants. A low 1-minute Apgar score (≤3) may increase the risk of death in very preterm infants, while high gestational age, high birth weight, and prenatal use of glucocorticoids are associated with the reduced risk of death. A low 5-minute Apgar score (≤3) and maternal chorioamnionitis may increase the risk of serious complications in these infants, while high gestational age and high birth weight may reduce the risk of serious complications.

Bioassay-guided isolation of two antifungal compounds from Magnolia officinalis, and the mechanism of action of honokiol.

Magnolia officinalis, as a well-known herb worldwide, has been widely used to treat multiple diseases for a long time. In this study, the petroleum ether extract from M. officinalis showed effective antifungal activity against seven plant pathogens (particularly against R. solani with an inhibition rate of 100.00% at 250 µg/mL). Honokiol and magnolol, isolated by the bioassay-guided method, exhibited greater antifungal activity than tebuconazole (EC50 = 3.07 µg/mL, p ≤ 0.001) against R. solani, which EC50 values were 2.18 µg/mL and 3.48 µg/mL, respectively. We used transcriptomics to explore the mechanism of action of honokiol against R. solani. Results indicated that honokiol may exert antifungal effects by blocking the oxidative phosphorylation metabolic pathway. Further studies indicated that honokiol induced ROS overproduction, disrupted the mitochondrial function, affected respiration, and blocked the TCA cycle, which eventually inhibited ATP production. Besides, honokiol also damaged cell membranes and caused morphological changes. This study demonstrated that the lignans isolated from M. officinalis possess the potential to be developed as botanical fungicides.

Structural Simplification of Cryptolepine to Obtain Novel Antifungal Quinoline Derivatives against Phytopathogenic Fungi.

A series of quinoline derivatives were designed and synthesized by the structural simplification of cryptolepine and evaluated for their fungicidal activity against six phytopathogenic fungi. Most of these compounds exhibited remarkable activities against Botrytis cinereain vitro. Among them, compounds A18 and L01 showed superior antifungal activity. Significantly, compared to cryptolepine, compound A18 exhibited broad-spectrum inhibitory activities against B. cinerea, Sclerotinia sclerotiorum, Rhizoctonia solani, Phytophthora capsica, Magnaporthe oryzae, and Fusarium graminearum with the respective EC50 values of 0.249, 1.569, 3.915, 0.505, 0.246, and 4.999 µg/mL. Compound L01 displayed the best antifungal activity against B. cinerea with an EC50 value of 0.156 µg/mL. Preliminary mechanistic studies showed that compound A18 could inhibit spore germination, affect the permeability of the cell membrane, increase the content of reactive oxygen species, and affect the morphology of hyphae and cells. Moreover, compound A18 showed excellent in vivo protective effect against B. cinerea, which was more potent than pyrimethanil and equitant to cryptolepine. These results evidenced that compound A18 displayed superior fungicidal activities and could be a potential fungicidal candidate against plant fungal diseases.

Design, synthesis and biological evaluation of novel evodiamine and rutaecarpine derivatives against phytopathogenic fungi.

Evodiamine and rutaecarpine are two alkaloids isolated from traditional Chinese herbal medicine Evodia rutaecarpa, which have been reported to have various biological activities in past decades. To explore the potential applications for evodiamine and rutaecarpine alkaloids and their derivatives, various kinds of evodiamine and rutaecarpine derivatives were designed and synthesized. Their antifungal profile against six phytopathogenic fungi Rhizoctonia solani, Botrytis cinerea, Fusarium graminearum, Fusarium oxysporum, Sclerotinia sclerotiorum, and Magnaporthe oryzae were evaluated for the first time. Furthermore, a series of modified imidazole derivatives of rutaecarpine were synthesized to investigate the structure-activity relationship. The results of antifungal activities in vitro showed that imidazole derivative of rutaecarpine A1 exhibited broad-spectrum inhibitory activities against R. solani, B. cinerea, F. oxysporum, S. sclerotiorum, M. oryzae and F. graminearum with EC50 values of 1.97, 5.97, 12.72, 2.87 and 16.58 µg/mL, respectively. Preliminary mechanistic studies showed that compound A1 might cause mycelial abnormalities of S. sclerotiorum, mitochondrial distortion and swelling, and inhibition of sclerotia formation and germination. Moreover, the curative effects of compound A1 were 94.7%, 81.5%, 80.8%, 65.0% at 400, 200, 100, 50 µg/mL in vivo experiments, which was far more effective than the positive control azoxystrobin. Significantly, no phytotoxicity of compound A1 on oilseed rape leaves was observed obviously even at a high concentration of 400 µg/mL. Therefore, compound A1 is expected to be a novel leading structure for the development of new antifungal agents.

The risk factors and prognosis of bilateral primary breast cancer: a comparative study with unilateral breast cancer.

This study was performed to determine the risk factors and evaluate the outcome of bilateral breast cancer (BBC). We reviewed the records of 170 patients with BBC and 1,677 with unilateral breast cancer (UBC), and compared their personal history, histopatholgical characteristics, clinical findings, and treatment, and postoperative follow-up records. The patients with UBC were more likely to develop contralateral cancer with the features including: young age at onset, especially younger than 40, premenopause, late primiparity, breast cancer family history, benign mammary disease history, and a tumor larger than 5 cm (p < 0.05). After adjustment by multivariate analysis, we concluded that breast cancer family history and age at onset younger than 40 years old were the independent risk factors for BBC. There were no significant differences for distant metastasis or overall survival between BBC and UBC (p > 0.05). We observed that 64.1% of the second breast cancer occurred within 5 years after the operation of the first cancer, and medical examination could improve the early diagnosis of the contralateral breast cancer. Contrary to common belief, our study showed that BBC and UBC had similar biological features and prognosis (p > 0.05). The excessive treatment and prophylactic measures may be unnecessary in this seemingly aggressive breast cancer. The patients with UBC younger than 40 or with breast cancer family history should have intensive contralateral breast followup, especially within 5 years after in the initial treatment.

Risk factors for respiratory assistance in premature infants.

Premature infants are prone to dyspnea after birth due to immature development, and some infants require respiratory assistance. However, the risk factors for respiratory assistance in premature infants are rarely reported. The present study enrolled 3,394 premature infants (665 infants had been provided with respiratory assistance and 2,729 had not used respiratory assistance) to retrospectively analyze the risk factors associated with respiratory aid. The multivariate logistic regression analysis demonstrated that placental abnormality [odds ratio (OR)=1.284; P=0.048], the male sex (OR=0.696; P=0.001), delivery via cesarean section (OR=1.538; P<0.001), low 1-min Apgar score (OR=0.727; P<0.001), low birth weight (OR=0.999; P=0.005) and low gestational age (OR=0.616; P<0.001) were independent risk factors for respiratory assistance in premature infants. Overall, a number of risk factors, including placental abnormality, cesarean section, low 1-min Apgar score, low birth weight and small gestational age, were identified for respiratory assistance in premature infants. By conducting a risk assessment of risk factors at birth and using this information to provide timely respiratory assistance, the survival rates of premature infants may increase.

[Research on the postoperative complications and its risk factors of breast reconstruction with abdominal flaps].

OBJECTIVE: To study the postoperative complications and its risk factors in patients underwent breast reconstruction with abdominal flaps. METHODS: The clinical data of 115 cases underwent breast reconstructions with abdominal flaps from May 2001 to October 2008 was reviewed. The postoperative complications included total flap necrosis, partial flap necrosis, fat necrosis, hernia, bulge, fat liquefaction and infection. The risk factors of complication rates were also evaluated. RESULTS: The total postoperative complications rate was 17.4% (20/115). No severe complications was found, such as total flap necrosis, hernia and bulge. The most common complications of flap was fat necrosis which occurred in 6 cases (5.2%), partial flap necrosis in 5 cases (4.3%) and infection in 1 case (0.9%). The donor-site complications included fat liquefaction which occurred in 8 cases (7.0%) and infection in 3 cases (2.6%). No significant relation was found between patient's age, body mass index (BMI), timing of surgery and the postoperative complication rate. The postoperative complications occurred more frequently in active smokers, patients with radiotherapy history, or reconstructions with pedicled transverse rectus abdominis myocutaneous (TRAM) flaps. But no significant difference was found in those factors. CONCLUSIONS: Fewer complications happens in patients with a reconstruction with deep inferior epigastric perforator (DIEP) flap. Abdominal flap should be performed with more consideration in active smokers or patients with a radiotherapy history. Age and obesity should not be contraindications to breast reconstruction with abdominal flaps.

[Performances Analysis of an Upflow Anaerobic Filter for Domestic Sewage Treatment].

Upflow anaerobic filter (UAF) with actual domestic wastewater were examined in this study. The Impacts of hydraulic retention time (HRT) on the performance of a UAF and a primary methanogen group were investigated at mesophilic conditions. The chemical oxygen demand (COD) removal rate was more than 75% after 28 days acclimation at 35℃ and HRT of 24 h. With a gradual decrease in the HRT, the COD removal rate first increased and then decreased. When the HRT was 5 h, the COD removal rate was the highest, with an average 81.71% and a maximum of 87.18%. When the HRT decreased to 2.5 h, the average COD removal rate decreased to 75.12%. The methane produced per unit mass of substrate consumed (CH4/CODre) and volume fraction increased with a decrease in HRT. When the HRT was 2.5 h, it reached 0.30 L·g-1, and the volume fraction of methane was maintained at about 73%. The energy generated by the system met the energy demands of the peristaltic pump. Quantitative analysis of the primary methanogen group in the system indicates that Methanosarcinales is the dominant in the system. With a decrease in HRT, the abundances of acetoclastic and hydrogenotrophic methanogens increased significantly.

[Cox proportional hazard model analysis of prognosis in patients with carcinoma of esophagus and gastric cardia after radical resection].

OBJECTIVE: To investigate the factors affecting the long-term survival of patients with carcinoma of esophagus and gastric cardia after curative resection. METHODS: The clinical data of 906 patients with carcinoma of esophagus and gastric cardia treated by radical resection in 1996 - 2004 were analyzed retrospectively. Twelve clinicopathological factors possibly influencing survival were encoded and assessed by Cox regression analysis. RESULTS: The 1-, 3- and 5-year cumulative survival rates were 89.8%, 75.4% and 71.7%, respectively. The univariate analysis showed that age, length of tumor, pathological differentiation, number of metastatic lymph nodes, depth of invasion, involvement of adjacent organs and the TNM stage influenced the prognosis significantly (P < 0.01). However, multivariate analysis showed that pathologic differentiation, number of metastatic lymph nodes, involvement of adjacent organs and TNM stage were independent prognostic factors (P < 0.05). CONCLUSION: The independent prognostic factors of the patients with carcinoma of esophagus and gastric cardia are pathologic differentiation, TNM stage, number of metastatic lymph nodes, and involvement of adjacent organs. The other factors influencing survival are age, length of tumor and depth of invasion. Furthermore, invasion of adjacent organs suggests worse prognosis, and should be followed-up closely.

[The trend of clinical characteristics and prognosis of women's breast cancer 1981 - 2000].

OBJECTIVE: To investigate the trend of clinical characteristics and prognosis of breast cancer in women during the past 2 decades (1981 - 2000). METHODS: 1678 cases were sampled randomly from the 11400 breast cancer patients who were hospitalized in Tianjin Medical University Cancer Hospital 1981 - 2000. Retrospective analysis of the clinical characteristics and prognosis between different eras was conducted. RESULTS: The numbers of hospitalized female breast cancer patients during the periods 1981 - 1985, 1986 - 1990, 1991 - 1995, 1996 - 2000 were 209, 316, 449, and 704 respectively with an increase of 237% within 20 years and an annual increase rate of 11.8%. The mean onset age remained 46 - 47. The detection rate of breast cancer at early stages (stages 0 approximately I) was 15.5% during 1996 - 2000, higher than that during 1981 - 1985 (12%), and the detection rate of breast cancer at stage III during 1996 - 2000 was 16.5%, lower than that during 1981 - 1985 (24.9%). The proportion of carcinoma in situ during 1996 - 2000 was 4.4%, higher than that during 1981 - 1985 (3.7%). The axillary lymph node positive rate during 1996 - 2000 was 46.3%, lower than that during 1981 - 1985 (51.8%s), and the local recurrence/distance metastasis rate during 1996 - 200 was 13.5%, lower than that during 1981 - 1985 (27.8%). The 5-year and 10-year survival rates during 1996 - 2000 were 86% and 74%, both higher than those during 1981 - 1985 (76% and 63% respectively). CONCLUSION: During the past 2 decades, the incidence of breast cancer shows an uptrend. The proportion of early stage breast cancer increases gradually and the proportion of axillary lymph node positive rate and metastasis rate show a downtrend. The prognosis of the breast cancer patients has been improved during these periods.

Substrate specificity of human kallikreins 1 and 6 determined by phage display.

The human tissue kallikrein (KLK) family contains 15 secreted serine proteases that are expressed in a wide range of tissues and have been implicated in different physiological functions and disease states. Of these, KLK1 has been shown to be involved in the regulation of multiple physiological processes such as blood pressure, smooth muscle contraction, and vascular cell growth. KLK6 is overexpressed in breast and ovarian cancer tissues and has been shown to cleave peptide derived from human myelin protein and Abeta amyloid peptide in vitro. Here we analyzed the substrate specificity of KLK1 and KLK6, by substrate phage display using a random octapeptide library. Consistent with earlier biochemical data, KLK1 was shown to exhibit both trypsin- and chymotrypsin-like selectivities with Tyr/Arg preferred at site P1, Ser/Arg strongly preferred at P1', and Phe/Leu at P2. KLK6 displayed trypsin-like activity, with the P1 position occupied only by Arg and a strong preference for Ser in P1'. Docking simulations of consensus peptide provide information on the identity of the enzyme residues that are responsible for substrate binding. Bioinformatic analysis suggested several putative KLK6 protein substrates, such as ionotropic glutamate receptor (GluR) and synphilin.

[A seven-year follow-up analysis on prognosis factors of stroke patients].

OBJECTIVE: To describe survival conditions of patients with stroke and to analyze the factors associated with survival, a seven-year follow-up study was carried out. METHODS: From Nov. 1995 to Dec. 1996, 189 stroke patients were selected in the Department of Neurology, General Hospital, Tianjin Medical University. Cases were followed up since the onset of stroke. Data collected would include case history, illness and survival conditions. Kaplan-Meier methods were used for survival description. Cox regression was used for prognostic factors analyses. RESULTS: A total number of 82 patients had been dead during the period of study and among them,58 cases died from stroke. The survival rate was 79.86% in one year, 65.46% in three years and 57.46% in seven years. Factors with statistical significance that associated with survival would include: age (RR = 1.065, P < 0.001), physical exercises before stroke (RR = 0.308, P<0.001), hypertension history (RR = 1.785, P < 0.05) and stroke history (RR =2.493, P < 0.001) while factors associated with severity of the illness were: area of cerebral lesion, conditions when discharged from the hospital, rehabilitative treatment of post-discharge and recurrence. We also found that social-psychical factors as rehabilitative confidence, repression, negative event, support from relatives and friends were related to survival of stroke. CONCLUSIONS: Histories on hypertension, stroke and brain injury condition were related to the rate survival on stroke. Patients persisting physical exercises before stroke had better prognosis. The survival rate of patients with recurrence was lower than those without while social psychic factors might be related to survival.