Taxa-4(5),11(12)-diene is the precursor for paclitaxel biosynthesis. The diterpenoid paclitaxel (marketed as Taxol), a plant secondary metabolite isolated from yew, is an effective drug widely used in the treatment of numerous cancers. However, further application of taxol has been restricted due to its low yield in plants and the difficulties in extraction. To increase the intact isoprene flux, we constructed the fusion gene plasmid pBgGGTS and individual cassette plasmid pBgGGgTS to enhance the expression levels of geranylgeranyl diphosphate synthase gene (ggpps) and a taxadiene synthase gene (ts) in Coprinopsis cinerea. These two plasmids were separately transformed into C. cinerea LT2 strain, resulting in several putative transformants. Putative transformants were determined by PCR technique, indicating that 5 out of 13 putative transformants transformed by pBgGGTS and 6 out of 13 putative transformants transformed by pBgGGgTS, respectively. Additionally, the Southern blotting analysis of these 10 transformants confirmed that both ggpps and ts gene were stably integrated into the genome of C. cinerea. Crude extracts from each of the transformants were analyzed. There is no difference in the mycelium extracts among the wild-type LT2 and two types of transformants. However, analysis of culture filtrates indicated that an additional GC peak was found at the retention time of 16.762 min which was absent in the wild type control. The mass fragmentation pattern of this peak had the same diagnostic ions with taxa-4(5),11(12)-diene. According to peak area, the amounts of taxa-4(5),11(12)-diene in each fermented broth were 44 ng/L (transformed with pBgGGgTS) and 30 ng/L (transformed with pBgGGTS), respectively. In conclusion, co-expression of the ggpps and ts gene could increase the taxadiene production in C. cinerea.
Agaricales ; metabolism ; Alkenes ; metabolism ; Diterpenes ; metabolism ; Farnesyltranstransferase ; genetics ; metabolism ; Genetic Engineering ; Isomerases ; genetics ; metabolism ; Paclitaxel ; Plasmids

Objective To observe the different changes of macular microstructure in patients with large idiopathic macular hole (IMH) treated with vitrectomy combined with internal limiting membrane (ILM) transplantation or not.Methods Forty eyes in 40 consecutive patients with giant IMH (≥500 μm) were included in the study.Twenty eyes received vitrectomy with ILM transplantation (ILM transplantation group) and others with ILM peel off (ILM removal group).During the operation,a proper size of the ILM was removed and filled in the bottom of the macular hole.The age,duration of disease and the ocular laterality of the two groups of patients were not statistically significant (P＞0.05).Minimum resolution angle in logarithmic (logMAR) best corrected visual acuity (BCVA) and frequency domain optical coherence tomography (SD-OCT) scan were examined.There was no statistically significant difference in logMAR BCVA,average defect diameter of photoreceptor ellipsoid (IS/OS) and average defect diameter of external limiting membrane (ELM) between two groups (t=0.128,1.452,1.321;P＞0.05).The logMAR BCVA and SD-OCT were examined on 1,3,6,12 months postoperatively.Results On 1 month after the surgery,there was no statistically significant difference in logMAR BCVA,average defect diameter of IS/OS and average defect diameter of ELM between two groups (t=1.226,1.435,1.018;P＞0.05).On 3,6,12 months after the surgery,compared with ILM removal group,the logMAR BCVA (t=2.059,2.871,2.415) increased and the average defect diameter of IS/OS (t =2.070,2.110,2.121) and ELM (t =2.034,3.647,3.556) significantly reduced in ILM transplantation group (P＜0.05).On 1 month after the surgery,there was statistically significant difference in CRT between two groups (t=2.113,P＜0.05).On 3,6,12 months after the surgery,there was no statistically significant difference in CRT between two groups (t=0.428,0.847,0.849;P＞0.05).Conclusion Compared with vitrectomy combined with ILM peeling surgery,the diameter of IS/OS and ELM defect were significantly decreased after vitrectomy combined with ILM transplantation in the patients with large IMH.

BACKGROUNDTo evaluate the activity and toxicity of paclitaxel as second-line treatment for advanced non-small cell lung cancer (NSCLC).

METHODSForty patients with recurrent advanced NSCLC were enrolled. Thirty-six patients were managed with regular regimen. Paclitaxel 135 mg/m², 3 h, on day 1; DDP 75 mg/m² or carboplatin 300-350 mg/m² on day 2. Four patients were managed with weekly regimen. Paclitaxel 60 mg/m²,3 h, on days 1,8,15; DDP 75 mg/m² on day 2. It was repeated every three or four weeks, up to two to four cycles.

RESULTSThirty-six cases were evaluated for response and 27 for survival. The objective response rate was 13.9% (5/36). At least one tumor-related symptom relief was observed in 21 patients (58.3%). The median survival duration was 26.4 weeks and 1-year survival rate was 8% (4/36). The main toxicities included myelosuppression, fatigue and myalgia-arthralgia neuropathy.

CONCLUSIONSPaclitaxel has advantage to be well-tolerated and improve tumor-related symptom. Further studies with standardization of dose and regimen will be needed to clarify its role in the second-line treatment.

Background Giant idiopathic macular hole (IMH) severely affects visual acuity and increases operative difficulty during the surgery,so modifying or optimizing the operation method is helpful for improving the prognosis.Objective This study aimed to evaluate the efficacy and safty of vitrectomy combined with free internal limiting membrane transplantation for large IMH.Methods A prospective serial cases-observational study was carried out under the informed consent of each patient.Forty-two eyes of consecutive 42 patients with IMH of mean diameter (814.31±112.95)μm were included in the Second Hospital of Hebei Medical University from January 2013 to November 2015.All the eyes received vitrectomy combined with free internal limiting membrane transplantation and 12％ C3F8 filling.The best corrected visual acuity (BCVA) (LogMAR),inner segment/outer segment (IS/OS) defect range,external limiting membrane defect range,retinal thickness at macular fovea were measured with slit lamp microscope,indirect ophthalmoscope and spectral-domain optical coherence tomography (SD-OCT) before surgery and 1,3,6 and 12 months after surgery.Results IMH complete closure in 97.6％ eyes (41/44) at 12 months after surgery.BCVA was improved after operation and showed a significant difference among various time points (F =28.032,P＜0.001).The IS/OS defect range was (1 112.00±45.44),(859.00±84.55),(649.00±52.47),(486.00±46.88) and (320.00±45.13) μm before surgery and 1,3,6 and 12 months after surgery,showing a significant difference among different time points (F=38.761,P＜0.001),and the IS/OS defect range was gradually shrinked after operation compared with that before operation (all at P＜0.05).The mean defect range of external limiting membrane was (1 038.00 ±39.63),(748.00±64.12),(585.00±48.88),(438.00±42.84) and (265.00±28.97)μm before surgery and 1,3,6 and 12 months after surgery,with a significant difference among various time points (F=36.459,P＜0.001),and the mean defect range of external limiting membrane was evidently reduced (all at P＜0.05).The foveal retinal thickness value was increased at 3,6 and 12 months after surgery,which was significantly higher than that 1 month after surgery (all at P＜0.05).The high reflect signal was faded away at 3 months after operation,indicating that implanted internal limiting membrane was decomposed and metabolized.Conclusions Vitrectomy combined with autologous internal limiting membrane transplantation seems to be safe and effective for large IMH.

BACKGROUNDTo evaluate the efficacy, side-effects, median survival duration and survival rate of vinorelbine (NVB) combined with cisplatin (DDP) in the treatment of non-small cell lung cancer (NSCLC).

METHODSA total of 220 patients with inoperable NSCLC received NVB and DDP combined chemotherapy: NVB 25-30 mg/(m²*d) on days 1 and 5 (or 8), DDP 60-80 mg/(m²*d) on day 2. The schedule was repeated every 28 days. The efficacy and side-effects were analysed and followed-up after at least two cycles of chemotherapy.

RESULTSThe overall response rate (CR+PR) was 30.9% (68/220). The response rate was 31.3% (51/163) in initial treatment group, and 29.8% (17/57) in retreatment group. The median survival duration was 8.3 months. The 1-, 2- and 3-year survival rates were 39.23%, 19.31% and 6.32%, respectively. The main side-effects were myelosuppression and digestive tract reactions.

CONCLUSIONSVinorelbine plus cisplatin is an effective and well-tolerated regimen for non small cell lung cancer and myelosuppression is its dose-limiting toxicity.

Objective:To explore the relationship between thyroid parenchymal echogenicity abnormalities and thyroid function evolution in health examination population.Methods:It was a cohort study. According to the inclusion and exclusion criteria, participants were selected from individuals who underwent thyroid color ultrasound and thyroid function tests at the Health Management Center of Tianjin Medical University General Hospital from January to December 2017. Data including age, gender, smoking history, alcohol consumption history, personal medical history, and thyroid function re-examination information were collected. Follow-up was conducted until the occurrence of thyroid function abnormalities or until the end of the follow-up period. The cumulative incidence rate and incidence density of thyroid function abnormalities were evaluated. Cox regression analysis was used to analyze the relationship between thyroid parenchymal echogenicity abnormalities and thyroid function evolution.Results:A total of 6 754 participants were included in this study, with an average age of (45.80±12.12) years, and females accounted for 42.7%. The mean follow-up time was 1.82 years, with a cumulative follow-up duration of 12 263 person-years. During the follow-up period, 154 new cases of thyroid function abnormalities occurred, with a cumulative incidence rate of 2.28% (95% CI: 1.94%-2.66%) and an incidence density of 12.56/1 000 person-years (95% CI: 10.66/1 000-14.69/1 000 person-years). The multivariate adjusted Cox regression analysis showed that individuals with thyroid parenchymal echogenicity abnormalities had a significantly increased risk of developing isolated thyroid stimulating hormone abnormalities and thyroid function abnormalities, with HR(95% CI) of 3.09 (2.02-4.73) and 2.92(1.96-4.33), respectively, both P<0.001. Stratified analysis showed that, except for body mass index <18.5 kg/m 2, current smoking, and current alcohol consumption, all other stratified factors showed a significant increase in the risk of thyroid function abnormalities in individuals with thyroid parenchymal echogenicity abnormalities. Conclusion:Thyroid parenchymal echogenicity abnormalities are important risk factors for the evolution of thyroid function abnormalities. Continuous monitoring of thyroid function should be given due attention.