Anidulafungin is an effective antifungal medicine, which can inhibit activities of candida in vitro and in vivo. Echinocandin B (ECB) is the key precursor of Anidulafungin, thus the price and market prospect of Anidulafungin is directly due to the fermentation titer of ECB. In this study, Aspergillus nidulans was used for ECB fermentation, and the influence of adding microparticles on ECB fermentation was studied, such as talcum powder, Al2O3, and glass beads. The particle size and concentration were the key factors for mycelium morphology and ECB production, and ECB production could reach 1 262.9 mg/L and 1 344.1 mg/L by adding talcum powder of 20 g/L (d50 = 14.2 μm) and 7 glass beads (6 mm), an increase by 33.2% and 41.7%, respectively. The results indicated that the mycelium morphology of filamentous microorganisms and the product yield of fermentation could be improved by adding microparticles remarkably, and it provide an important method for the fermentative optimization of filamentous microorganisms.
Antifungal Agents ; metabolism ; Aspergillus nidulans ; metabolism ; Echinocandins ; biosynthesis ; Fermentation ; Fungal Proteins ; biosynthesis ; Industrial Microbiology ; methods

Objective To investigate the efficacy and safety of warfarin anticoagulation in Chinese elderly patients based on vitamin K epoxide reductase complex 1 (VKORC1) and cytochrome P450 2C9 (CYP2C9) genetic polymorphisms.Methods Clinical data of 41 elderly patients with initial anticoagulation therapy in our emergency department and respiratory department were collected.Patients were divided into observation group (n=20,patients treated with warfarin based on genetic polymorphisms) and control group (n =21,patients treated based on clinical experience).The international normalized ratio (INR),the time of INR stabilized within target range (2.0-3.0) and the incidence of bleeding episodes in 6-month follow up were compared between groups.Results INR within target range at day 3,4,5 and 7 were 0.0％,42.1％,52.6％,68.4％ in observation group and 0.0％,10.0％,25.0％,35.0％ in control group,respectively.There were significant differences in INR within target range at day 4,7 between the two groups (both P＜0.05),while no significant difference was found in INR within target range at day 5 (P＞0.05).The time of INR stabilized within target range was shorter in observation group than in control group [(9.5±2.4) d vs.(12.3± 4.8) d,P＜0.05].Bleeding complication occurred in 3 patients in observation group and 5 patients in control group,and there was no significant difference between the two groups.Conclusions Warfarin therapy based on VKORC1 and CYP2C9 gene polymorphisms may shorten the time of first INR reaching the target value and INR within target range in elderly patients.However,the risk of bleeding complications should be alerted.

Objective To investigate the effect of tripterygium glycosides (TG) contained serurn on the pathological boneforming related inflammatory markers and miR-21.Methods Previous isolated and cultured AS fibroblasts were stimulated using IL-1 of 1ng/ml for 24h,different concentrations of blank serum (5％,10％,and 15％) and TG contained serum (5％,10％,and 15％) were added for 48h.PGE-2,IL-17,IL-22,IL-23,CCL19 and CCL21 proteins were examined by Western blot.The osteogenesis marker BMP and microRNA-21 mRNAs were tested.Results 48 h after intervention,the expressions of inflammatory markers were obviously inhibited by TG contained serum;the boncforming related inflammatory markers,expressions of BMP-2 and miR-2t were all inhibited in a dose-dependent manner.Conclusions TG could inhibit the expressions of boneforming related inflammatory markers,BMP-2 and miR-21,thus providing theoretical basis to treat AS pathological boneforming.

Objective To evaluate the feasibility and outcome of laparoscopic cholecystectomy as a overnight procedure in China.Methods The data of 59 consecutive patients who had undergone outpatient laparoscopic cholecystectomy between Januaiy 2013 and January 2015.All the patients were operated in the morning hours and discharged during 24 hours after operation.They were contacted by telephone 3 days subsequent to surgery and were seen in the outpatient unit 7 days after.Results Fifty-nine laparoscopic cholecystectomies were performed.No Conversion to open surgery case.The average operation time was 25 minutes,and restore semi-liquid diet 6 hours after the operation All the 59 patients were discharged during 24 hours after operations.None of the patients had an emergency readmission.None of the patients had complications 7 days after discharged.The average payment was 8 240 yuan.Conclusion These results suggest that laparoscopic cholecystectomy can be routinely performed as a overnight procedure.

Objective To investigate the effect of sterol regulating element binding protein (SREBP1c) and its ac-tive form (SREBP1cm) on human protein kinase R-like endoplasmic reticulum kinase (PERK).Methods Re-porter victors of PERK promoter and its truncations were constructed with pGL 3-basic and co-transfected with internal reference pRL-SV40 into cell and luciferase activity was detected .pcDNA3.1 ( -)-SREBP1c or pcDNA3.1 ( -)-SREBP1cm was co-transfected with PERK promoter transcriptional activity core regions and the detection of dual -lu-ciferase reporter gene was used to analyze the regulation of SREBP 1c/1cm on PERK promoter transcriptional activity . The expression level of PERK mRNA and protein were detected by RT-PCR and Western blot .Results PERK pro-moter and truncations were successfully constructed into pGL 3-basic, and PERK promoter core area of transcription-al activity had determined;Dual-luciferase report gene showed that SREBP 1c inhibited PERK promoter transcrip-tional activity and SREBP1cm promoted PERK promoter transcriptional activity .RT-PCR and Western blot showed that SREBP1c decreased PERK mRNA and protein expression , but SREBP1cm increased PERK mRNA and protein expression, which was consistent with the detection of dual-luciferase report gene .Conclusions SREBP1c and SREBP1cm have a opposite regulation effect on PERK promoter transcriptional activity and its expression .

Objective To investigate the safety and feasibility of reoperatively laparoscopic technique in treatment for locally recurrent rectal cancer.Methods The study enrolled 17 patients with locally recurrent rectal cancer between February 2004 and September 2009 from Shanghai Minimally Invasive Surgery Center.The patients were divided into two groups according to their pelvic recurrence types:central recurrence group (n =14) and anterior recurrence group (n =3).Demographic,surgical data and survival outcomes between two groups were compared.Results The outcomes of demographic data between two groups were not different(P＞ 0.05 ).Compared with central recurrence group,anterior recurrence group had longer operating time (P =0.028).However,the differences of operative blood loss,complications,postoperative rehabilitative outcomes and ratio of R0 resction between groups were not significant ( P ＞ 0.05 ).The overall 5- year survival rate of all the patients was 36％.And the median survival time was 42 months without significant difference between two groups (x2 =1.641,P =0.200).Conclusions Reoperatively laparoscopic technique in treatment for locally recurrent rectal cancer is safe and feasible.Selected patients,specialist operation and higer ratio of R0 resection are the key factors conducive to better short-and long-term outcomes.

Objective To compare the safety and efficiency of laparoscopic transabdominal preperitoneal (TAPP) and totally extraperitoneal (TEP) procedure for herniorrhaphy. Methods Between Jan 1997 and Jan 2005, 235 patients (274 hernias) underwent laparoscopic inguinal hernia repair with 163 TAPP in 139 patients and 111 TEP procedures in 96 patients. Postoperative patients were followed-up for 6-24 months. Results No conversion was necessary. There were no significant differences between TAPP and TEP when considering operating time, postoperative hospital stay, VAS and time to return to daily activity. The recurrence rate was 1. 8% and 0. 9% respectively in TAPP and TEP( P = 0. 524). The total incidence of postoperative complications were 15. 3% and 10. 8% respectively ( P = 0. 282 ) , with seroma, transient neurapraxia and urinary retention being 5. 5% vs. 4. 5% ( P = 0. 707 ) , 5. 5% vs. 3. 6% ( P = 0.464) and 3. 1% vs. 2.7%(P = 1.000). The inhospital cost was significantly higher in TAPP than in TEP(P=0.000). Conclusion TAPP and TEP are both safe and efficient tension-free technique. Surgeons' experience was more important than choice of approach to laparoscopic repair for groin hernias.

BackgroundHuman retinal gliocytes play an important role in proliferative diseases,which are the basis of in vitro studies.Researchers have cultured human retinal gliocytes in the past.In our study,we found that the cells we cultured presented a unique shape different from those by other researchers.ObjectiveThis study was to design to produce a new culture and purification method for retinal gliocyte in vitro.Methods Retinal tissue was isolated from human eyeballs and digested using the two-step digestion method (2％ pancreatin and 0.133％collagenase Ⅵ) to harvest the retinal glio cytes.The cells were collected and cultured in endothelial cell-targeted nutrient culture containing 10％ fetal calf serum and supplemented with β-endothelial cell growth factor (ECGF) and sodium heparin,and the culture dishes were coated with fibronectin(FN) to promote the attachment of retinal gliocyte.During the culturing process,the gliocytes were identified by the observation of morphological characteristic and regular histological examination.The identification of the cells also was performed by immunochemistry targeting glial fibrillary acidic protein (GFAP),Vimentin,neuron specific enolase ( NSE ),S-100,CD34,and Ⅷ factor.Results Retinal gliocytes were isolated successfully from the human retina by the two-step digestion method.Primary cultured cells attached after 72 hours and achieved confluency between day 9 and 10 that were aligned petaliform in shape.Regular histological examination after H&E staining showed blue cell nuclei and light red cytoplasm.The target cells presented with strong responses for GFAP and Vimentin and no response for NSE,S-100,CD34 and Ⅷ factor.ConclusionsLarge amount of purified human retinal gliocytes can be obtained by two-step digestion and cultured in endothelial cells-targeted culture medium supplemented with β-ECGF and sodium heparin in plates coated with FN.The cultured cells expressed markers for retinal gliocytes.However,specific features of these cells remain to be further elucidated.

AIM: To investigate the expression of connective tissue growth factor (CTGF) and α-SMA in human lens epithelium cell (HLEC) line B3 after transfection by liposome-coated siRNA targeting CTGF.METHODS: HLECs were transfected with small interfering RNA (siRNA) targeting CTGF,labeled with 5`-fluorescein isothiocyanate (5`-FITC) and coated with lipofectamine.The transfection ratio was evaluated via fluorescence intensity.Cell counting kit-8 (CCK-8) assay was performed to assess cytoviability of both non-transfected and transfected HLECs.Quantitative RT-PCR,cell immunochemistry and Western blot analysis were conducted to detect the expression changes of CTGF and α-SMA after transfection.RESULTS: A highly effective transfection ratio was observed in siRNA co-transfected with lipofectamine.The transfection ratio reached 95% at 24h.The proliferation of HLECs was inhibited by siRNA after 72h transfection.The expression of CTGF and α-SMA significantly decreased in HLECs after transfected by CTGF siRNA for 24h.This effect was not found in negative control siRNA.CONCLUSIONS: SiRNA targeting CTGF decreased CTGF and α-SMA expression in HLECs,which is a potential therapeutic strategy for posterior capsular opacification.

Objective Pemetrexed and β-elemene can inhibit the growth of tumor cells .This study was to investigate the effect of pemetrexed combined with β-elemene on the proliferation and apoptosis of cervical cancer HeLa cells. Methods Cervical cancer HeLa cells were treated with pemetrexed at the concentrations of 38, 76, 152, 228, and 304μg/mL, and at 24 and 48 hours of treatment subjected to MTT for detection of their proliferation .The experiment included four groups , with the cells treated with β-elemene ( 125μg/mL) , pemetrexed ( 76 μg/mL ) , β-elemene ( 125 μg/mL ) +pemetrexed (76μg/mL), and nothing (blank control) for 24 hours, followed by determination of their proliferation and apoptosis by MTT and flow cytometry, respectively. Results Pemetrexed at 38, 76, 152 and 228μg/mL inhibited the proliferation of the HeLa cells in a concentration-dependent manner, with the inhibition rates of (7.24 ±3.78), (7.94 ±4.37), (11.10 ±2.86) and (15.88 ± 3.38)%at 24 hours, and (16.69 ±0.95), (22.54 ±1.53), (24.48 ±0.92) and (25.54 ±3.61)%at 48 hours, both with statis-tically significant differences between any two groups (P<0.05).Significant differences were also found in the proliferation rate of the same concentration of pemetrexed at the two time points (P<0.05).The combination of pemetrexed and β-elemene showed an inhibi-tion rate of (49.95 ±5.76)%at 24 hours, remarkably higher than (24.36 ±5.59)%in theβ-elemene group and (10.69 ±1.37)%in the pemetrexed group (P<0.01). Conclusion Pemetrexed combined with β-elemene can significantly inhibit the proliferation and synergistically accelerate the apoptosis of HeLa cells .