Genetic interactions between the Golgi Ca2+/H+ exchanger Gdt1 and the plasma membrane calcium channel Cch1/Mid1 in the regulation of calcium homeostasis, stress response and virulence in Candida albicans.

The Golgi-localized Saccharomyces cerevisiae ScGdt1 is a member of the cation/Ca(2+) exchanger superfamily. We show here that Candida albicans CaGdt1 is the functional homolog of ScGdt1 in calcium sensitivity, and shows genetic interactions with CaCch1 or CaMid1 in response to ER stresses. In addition, similar to ScCCH1 and ScMID1, deletion of either CaCCH1 or CaMID1 leads to a growth sensitivity of cells to cold stress, which can be suppressed by deletion of CaGDT1. Furthermore, deletion of CaCCH1 leads to a severe delay in filamentation of C. albicans cells, and this defect is abolished by deletion of CaGDT1. In contrast, CaGDT1 does not show genetic interaction with CaMID1 in filamentation. Interestingly, C. albicans cells lacking both CaMID1 and CaGDT1 exhibit an intermediate virulence between C. albicans cells lacking CaCCH1 (non-virulent) and C. albicans cells lacking CaGDT1 (partially virulent), while C. albicans cells lacking both CaCCH1 and CaGDT1 are not virulent in a mouse model of systemic candidiasis. Therefore, CaGdt1 genetically interacts with the plasma membrane calcium channel, CaCch1/CaMid1, in the response of C. albicans cells to cold and ER stresses and antifungal drug challenge as well as in filamentation and virulence.

Genetic interactions between Rch1 and the high-affinity calcium influx system Cch1/Mid1/Ecm7 in the regulation of calcium homeostasis, drug tolerance, hyphal development and virulence in Candida albicans.

The high-affinity calcium influx system (HACS) consisted of CaCch1, CaMid1 and CaEcm7 controls calcium influx into the cell in response to environmental stimuli. The plasma membrane protein CaRch1 is a negative regulator of calcium influx in Candida albicans. In this study, we show that deletion of any of the HACS components suppresses the calcium hypersensitivity of, and the elevated activation level of calcium/calcineurin signaling in, C. albicans cells lacking CaRCH1. In contrast, CaRCH1 is epistatic to the HACS system in the tolerance of antifungal drugs. In addition, cells lacking CaRCH1 are sensitive to tunicamycin, show a delay in in vitro filamentation and an altered colony surface morphology, and are attenuated in virulence in a mouse systemic model. Cells lacking CaCCH1 and CaMID1, but not CaECM7, are sensitive to tunicamycin. Deletion of CaRCH1 increases the tunicamycin sensitivity of cells lacking CaECM7 or CaMID1, but not CaCCH1. Furthermore, deletion of CaRCH1 suppresses the defect in hyphal development due to the deletion of CaCCH1 or CaECM7, and increases the virulence of cells lacking any of the HACS components. Therefore, CaRch1 genetically interacts with the HACS components in different fashions for these functions.

Exosomes derived from mesenchymal stem cells containing berberine for ulcerative colitis therapy.

Berberine (Ber), an isoquinoline alkaloid, is a potential drug therapy for ulcerative colitis (UC) because of its anti-inflammatory activity, high biological safety, and few side effects. Nevertheless, its clinical application is hindered by its limited water solubility and low bioavailability. Currently, compared to synthetic nanocarriers, exosomes as carriers possess advantages such as low toxicity, high stability, and high specificity. Human placental mesenchymal stem cell-derived exosomes (HplMSC-Exos) have emerged as a promising drug delivery system, offering intrinsic anti-inflammatory and antioxidant activities. Therefore, we engineered MSC-Exos loaded with Ber (Exos-Ber) to enhance the solubility and bioavailability of Ber and for colon targeting, revealing a novel approach for treating UC with natural compounds. Structurally and functionally, Exos-Ber closely resembled unmodified Exos. Both in vitro and in vivo investigations confirmed the antioxidant and anti-inflammatory properties of Exos-Ber. Notably, Exos-Ber exhibited reparative effects on injured epithelial cells and reduced cellular apoptosis. Furthermore, Exos-Ber concurrently demonstrated anti-inflammatory and antioxidant activities, contributing to the mitigation of UC, possibly through its modulation of the MAPK signaling pathway. Overall, our findings demonstrate the potential of Exos-Ber as a promising therapeutic option for alleviating UC, highlighting its capacity to enhance the clinical applicability of Ber.

[Effects of ulinastatin on coagulation function and deep vein thrombosis in patients undergoing hip joint replacement].

OBJECTIVE: To investigate the effects of ulinastatin (Uti) and low-molecular-weight heparin (Lmwh) on coagulation function and deep vein thrombosis (DVT) in patients undergoing hip joint replacement. METHODS: From March to December 2010 150 ASAI-II patients with average age of 72.5 (65 - 85) years undergoing hip joint replacement were randomly divided into 3 groups (n = 50 each): normal saline (NS) control group (Group C), Uti group (Group U) and Lmwh group (Group L). Group U received intravenous infusion of ulinastatin (10 000 U/kg) at preoperative, perioperative and after operation 1, 2 and 3 d, respectively. Group C received the same volume of NS instead of Uti. Group L were injected Lmwh subcutaneously (3200 U/d) at preoperative, after operation 1, 2 and 3 d. Blood samples were taken before operation (T(0)), at the end of surgery (T(1)), 1 d (T(2)), 2 d (T(3)) and 3 d (T(4)) after operation for determination the values of R, K, α angle, MA and CI, using thromboelastography, and the DVT were also examined through color Doppler ultrasonography at 3 d after operation. RESULTS: Compared with T(0), R, K were shorter, α angle, MA and CI were larger in group C, the values at T(2) were up to the peak then declined at T(4). Compared with group C, the value of R, K were larger, the value of α angle, MA and CI were shorter in group U and group L. The DVT checked by ultrasonography were found in 20 cases in group C, 1 case in group U, and zero case in group L. The differences were no statistically significant between group U and group L. CONCLUSION: Intravenous infusion of Uti during the period of operation can correct the hypercoagulability of blood and decrease the incidence of DVT after operation.

[Effect of ulinastatin on thromboxane B2 and deep vein thrombosis in elderly patients after hip joint replacement].

OBJECTIVE: To determine the effect of ulinastatin on plasma thromboxane B(2) and deep vein thrombosis(DVT) in elderly patients after hip joint replacement. METHODS: Eighty ASAI-IIpatients aged 65-81 years undergoing hip joint replacement were randomly divided into 4 groups (n=20): Group U1 (ulinastatin 5 000 U/kg);Group U2 (ulinastatin 10 000 U/kg); Group U3 (ulinastatin 20 000 U/kg); and Group C (the same volume of saline as control).The blood samples were collected at 5 time points: preoperation (T(1)), immediately after the operation (T(2)), 1 d (T(3)), 2 d (T(4)) and 3 d after the operation (T(5)), respectively. Thromboxane B(2) was detected, and DVT was also examined through color Doppler ultrasonography 3 d after the operation. RESULTS: Compared with T(1), the level of thromboxane B(2) significantly increased in Group C at T(2)-5, in Group U1 at T(2-4), in Group U2 and U3 at T(2) (P<0.01). Compared with Group C, the concentration of thromboxane B(2) decreased in Group U1 at T(2-3), in Group U2 and U3 at T(2-4) (P<0.01). Compared with Group U1, thromboxane B(2) significantly decreased in Group U2 and U3 at T(2-4) (P<0.01).The incidence rate of DVT was 40% in Group C, 10% in Group U1. There was no incidence of DVT in the Group U2 and U3 (P>0.05). CONCLUSION: Ulinastatin can inhibit blood thromboxane B(2) level in dose dependent manner and prevent DVT in elderly patients after hip joint replacement.

Mechanism of the anti-inflammatory activity by a polysaccharide from Dictyophora indusiata in lipopolysaccharide-stimulated macrophages.

Dictyophora indusiata polysaccharides (DIP) shows antioxidant, anti-tumor and immunostimulatory activities. However, the anti-inflammatory roles of DIP in NLRP3 inflammasome in LPS-stimulated macrophages are still not well defined. In this study, we investigated the mechanism of the anti-inflammatory activity of DIP in LPS-primed RAW264.7 macrophages. Our data showed that DIP inhibited NF-κB signal pathway via modulating TLR4 expression, phosphorylation of IκBα and nuclear translocation of NF-κB-p65 subunit. Meanwhile, DIP reduced inflammasome activation via decreasing NLRP3 expression in cytoplasmic pools, limiting self-assembly of NLRP3 inflammasome, as well as the subsequent activation of caspase-1 and the secretion of IL-1ß and IL-18. For the first time, we show that the anti-inflammatory activity of DIP is mediated by inhibiting TLR4/NF-κB signal pathway and NLRP3 inflammasome activation during LPS-induced acute inflammation in RAW264.7 macrophages.

[Case-control study on the effect of ulinastatin on postoperative complications in elderly patients undergoing hip joint replacement].

OBJECTIVE: To evaluate the preventive effects of ulinastatin (Uti) on postoperative complications in elderly patients undergoing hip joint replacement. METHODS: From Angust 2009 to June 2010, 160 elderly patients undergoing selective hip joint replacement with ASA I to II were assessed according to American Society of Anesthesiologists classification, including 81 males and 79 females ranging in age from 65 to 83 years (mean 73.9 years). All the patients were divided into 2 groups according to random number table (80 patients in each group): control group (group C) and ulinastatin group (group U). The patients in Group U received intravenous injection of ulinastatin with a dose of 10,000 U/kg before skin incision,and then with dose of 5000 U/kg respectively at 1, 2 and 3 days after operatio. The patients in Group C received the same volume of normal saline instead of ulinastatin. Blood samples were taken preoperatively,at the end of surgery and 1, 2, 3 days after operation for determination of ALT, AST, Scr, BUN and Plasma D-dimer. Deep vein thrombosis and postoperative cognitive dysfunction (POCD) were also examined through color Doppler ultrasonography and neuroeognirive assessment on the postoperative 3 days respectively. RESULTS: Compared with the preoperative values, the contents of ALT, AST, Scr, BUN and plasma D-dimer in each group all increased. Compared with group C,the values of ALT, AST, Scr, BUN and plasma D-dimer decreased markedly (P < 0.05). The incidence rate of DVT and POCD was 0 and 3.75% in group U, which were lower than those of patients in the group C (40%, 27.5%) respectively. CONCLUSION: Intravenous infusion of ulinastatin during operation can protect important organ function, correct blood hypercoagulability, lower the occurrence of DVT and POCD, and prevent the postoperative complications in some degree.

[Effect of ulinastatin on post-operative cognition disorders in elderly patients undergoing hip joint replacement].

OBJECTIVE: To evaluate the effect of ulinastatin on post-operative Cognition disorders in elderly patients undergoing hip joint replacement. METHODS: Forty ASA I or II elderly patients undergoing selective hip joint replacement, aged > or = 65 years, were randomly divided into 2 groups (n = 40 each): control group and ulinastatin group. Ulinastatin group received iv infusion of ulinastatin (10,000 u/kg) after skin incision, (5,000 U/kg) after operation 1, 2, 3 d respectively, included 21 males and 19 females with an average age of (75.00 +/- 7.81) years old. Control group received the same volume of normal saline instead of ulinastatin, included 20 males and 20 females with an average age of (72.80 +/- 7.25) years old. Neuroeognitive testing was performed on the preoperative day and on the 3th postoperative day and post-operative cognition disorders was defined as 1 SD decline from baseline on neurocognitive assessment. Serum S100beta protein were measured before operation, at the end of surgery, 3, 24 h and 3 d after operation. RESULTS: The incidence rate of postoperative cognition disorders was 2.5% in ulinastatin group, there were lower than those of patients in the control group (25%) (P < 0.05); In control group, the scales for MMSE before and after operation were (25.2 +/- 2.1), (22.6 +/-2.5) scores and the level of serum S100beta protein at T0-4 were (0.041 +/- 0.012), (0.125 +/- 0.031), (0.178 +/- 0.036), (0.142 +/- 0.038), (0.048 +/- 0.015) microg/L. As well in ulinastatin group, above date were (25.9 +/- 2.4), (24.8 +/- 2.1), (0.040 +/- 0.013), (0.095 +/- 0.021), (0.116 +/- 0.017), (0.087 +/- 0.019) and (0.043 +/- 0.012) respectively. Compared with preoperative, MMSE evaluation scale was decreased on the 3th postoperative day and the S100beta was increased markedly at T1-3 in control group (P < 0.05); Compared with control group, MMSE evaluation scale was increased and the S100beta was decreased markedly at T1-3 in ulinastatin group (P < 0.05 ). CONCLUSION: Intravenous infusion of ulinastatin during operation can prevent the occurrence of POCD in elderly patients.