Lung metastasis is one of the most common metastases.Metastasectomy is only indicated for selected patients,and most patients are unsuited to surgery.The main treatment is systemic chemotherapy,however,the long-term survival is limited.With the development of precision radiotherapies such as threedimensional conformal radiation therapy (3D-CRT) and stereotactic body radiotherapy (SBRT),it is proved that radiotherapy is favorable for the patients with lung metastasis,especially for limited lung metastasis.SBRT can be obtained better survival,but still need large prospective studies.

1,2,4-Butanetriol (BT) is an important non-natural chemical with a variety of industrial applications. A recombinant Escherichia coli biosynthesizing BT from D-xylose was constructed by heterologously expressing xdh and mdlC, and knocking out competing pathway genes including xylA, xylB, yjhE, yagH and ycdW. To optimize BT synthesis pathway, the third catalytic step that catalyzes the decarboxylation reaction of 3-deoxy-D-glycero-pentulosonic acid was identified as a potential bottleneck. Consequently, 2-keto acid decarboxylases from three different microorganisms were screened, and the kivD gene from Lactococcus lactis was found to increase BT titer by 191%. The improved strain BW-025 reached a final BT titer of 2.38 g/L under optimized transformation conditions. Attempts on synthetic pathway optimization were also made by fine-tuning the expression levels of each enzyme involved in the whole pathway based on BW-025. As a result, an xdh overexpressed recombinant strain, BW-074 was finally generated, with 48.62% higher BT production than that of BW-025.
Butanols ; metabolism ; Escherichia coli ; metabolism ; Gene Knockout Techniques ; Genetic Engineering ; Industrial Microbiology ; methods ; Metabolic Networks and Pathways

Objective To investigate the effects of adhesion mediated by bone nlalTow stroma celh from leukemia patient on chemotherapeutics sensitivity and cell cycle of Jurkat cells in the co-cultured model.Methods Bone mw stroma cells were isohted and cultured from leukemia patients routinely.To construct the co-cultured model.Jurkat cells were co-cultured with BMSCs the irradiated layer by 60Co,and the model was observed with scanning electron microscope.The IC50 values of Jurkat cells expesured to DNR were quantified by MTT.The cell cycles of Jurkat cells after 24h-adhesion in the co-cultured model were analyzed by Facs.The expression of cyclin A,cyclin E and p27 in Jurkat cells adhered to BMSCs for 4h.24h and 48h were detected by Western blot.Results Jurkat ceUs in the co-cultured model showed a decreased sensitivity to DNR.ICSO values for normal BMSCs,leukemic BMSCs and non-adhered control were of 1.78Ixmol/L,2.30pznol/L and 0.45p,mol/L,respectively.The percentages of Go-Gl phase for leukemic BMSCs group and non-adhered control group were of 48.74％±8.77％and 27.83％壬1.86％.Respectively.The percentages of Gz-M phase for leukemic BMSCs group and non-adhered control group were of2.01％±1.17％and 20.33％±1.84％。Respectively.Compared with eomrol group,the 24h-ad- hesion mediated by BMSCs from leukemia patients up-regulated the percentage of Go-G1 phase of Jurkat cells(P

Objective To explore the feasibihty of TRAIL to be used to remove the leukemia cells from the autologous hemopoietic stem cell transplants. Methods The expression of decoy receptor 1 and decoy receptor 2 on the bone marrow mononuclear cell were routine-ly isolated and observed by fluorescence microscope after PE-DcR1 or PE-DcR2 stain. The apeptosis rates of mononuclearcell and Jurkat cells interfered by 200ng/ml TRAIL for 18h were determined by flow cytometry after AnnexinV/Pl stain. The interfered mononuclear cells were cultured to count the number to form colony-forming unit-fibroblast(CFU-F). The Jurkat cells labeled by green fluorescent protein were in-corporated into the mononuclear ceils and affected by 200ng/ml TRAIL for 24h. The incorporated cells were cultured in the system with GM-CSF and EPO and the numbers of the CFU-GM, BFU-E and fluorescence colony were counted on the seventh day. Results Both decoy re-ceptor land decoy receptor 2 of TRAIL can be detected on membrane or in cytoplasm of the bone marrow mononuclear cell. The apoptosis rate of mononuclear cell interfered by 200ng/ml TRAIL for 18h was (5.95±1.23)%, which was markedly lower than that of Jurkat cells (33.42±2.28) %. The number of CFU-F of TRAIL group and control group were 235.67 ~ 33.56 and 249.33±42.72, respectively. No marked difference can be found between the mentioned two groups. Moreover, TRAIL decreased the number of fluorescence colony formed by Jurkat cells without significant decreasing the number of CFU-GM and BFU-E formed by bone marrow mononuclear cells. Conclusion TRAIL can selectively induce apoptosis in Jurkat cells without marked toxic effect on the bone marrow mononuclear cells, which means that TRAIL can be used to remove the leukemia cells from the autologous hemopeietic stem cell transplants in vitro.

BACKGROUND:According to the thrust of document issued by State Drug Administration, the clinical experiment was carried onfufang duzhongjiangu keli (compound) (Bo Si Zhuang) in treatment of knee joint osteoarthritis.OBJECTIVE: To evaluate the improvement of the compound in treatment of knee joint osteoarthritis and its safety.DESIGN: Zhuanggu guanjie wan (bolus) was taken as controlled drug and double blind, double-simulation randomized method was designed.SETTING: Fujian Institute of Chinese Medicine, Guananmen Hospital of China Academy of Traditional Chinese Medicine, Institute of Orthopedics and Traumatology of China Academy of Traditional Chinese Medicine and Beijing Hospital of Chinese Medicine.PARTICIPANTS: Clinical experiment Ⅱ was performed since December 19, 1999, in which, 200 cases of knee joint osteoarthritis were observed and divided into compound group (100 cases) and bolus group (100 cases).From December 1999 to March 2000, clinical experiment Ⅲ was performed to observe 400 cases of knee joint osteoarthritis, in which, 300cases were divided in compound group and 100 cases in bolus group. All of cases were diagnosed by X-ray test and differentiated in Chinese medicine as insufficiency of liver and kidney and stasis of tendons and vessels. All of patients were in the known of experiment.METHODS: In compound group, fufang duzhong zhuanggu keli (1bag/time, 3 times/day) + simulated dosage of zhuanggu guanjie wan were administrated. In bolus group, fufang duzhong zhuanggu keli simulated dosage + zhuanggu guanjie wan (1bag/time, twice/day) were administrated.Double blind and double-simulation randomized control experiment was given in one-month treatment to observe clinical therapeutic effects.MAIN OUTCOME MEASURES: Evaluation on clinical indexes of joint function ,clinical therapeutic effect, syndrome score in Chinese medicine and adverse reaction.RESULTS: Totally 600 cases employed had all accomplished datum collections, no dropped-off case. ① The total effective rate of compound group was superior remarkably to bolus group (92.%, 82%). ② The result of joint function in compound group was superior remarkably to that of bolus group. ③ Concerning to improvement of syndromes in Chinese medicine, the result in compound group was superior to that of bolus group (the decreased integrals were 7.03±3.38 and 5.43±3.16 respectively). ④No obvious harmful effect presented during experiment.CONCLUSION: Fufang duzhong jiangu keli improves the symptoms of osteoarthritis of knee safely and effectively.

Objective To investigate the characteristics and risk factors for allogeneic transfusion after unilateral total knee arthroplasty (TKA).Methods 852 patients (663 female and 189 male) underwent primary unilateral TKA from January 2014 to December 2014 were included.Average age of included patients were 64.9±7.9 years old (22-87).829 patients were osteoarthritis,others rheumatoid arthritis.The ASA score,BMI,doctor groups,diabetes,hypertension,thrombus (duplex color Doppler ultrasonography),pre-HGB,pre-HCT,pre-TP,pre-Cr,pre-BUN,pre-PT,operation time,starting MABP of the operation,anesthesia and TXA were collected.Potential risk factors for allogeneic transfusion were analyzed statistically via univariate and multivariate regression analysis.Results The preoperative hemoglobin level in 71 (8.3％) patients were lower than that in normal (male ＜120 g/L,female ＜110 g/L).The hematokrit in 27 (3.2％) patients were lower than that in normal (male ＜40％,female ＜37％).TXA was used in 740 (86.9％) patients during the operation.Allogeneic transfusion was performed in 202 (23.7％) the patients after TKA.The differences in the following items within two groups were statistically significant via univariate analysis (P＜0.05),female and male,≥70 and ＜ 70 years,pre-HGB normal and low,pre-HCT normal and low and with and without TXA.Female [OR=2.283,95％CI (1.405,3.711)],patient age of 70 years or older [OR=2.048,95％CI (2.064,4.292)],preoperative hemoglobin level low [male ＜ 120 g/L,female ＜ 110 g/L,OR=1.506,95％CI (1.376,4.427)] and preoperative hematokrit below normal [male ＜ 40％,female ＜ 37％,OR=3.412,95％CI (1.086,6.591)] were independent predictors for postoperative allogeneic transfusion in multivariate regression analysis.Conclusion The allogeneic transfusion rate after unilateral TKA was 23.7％.Female,older than 70 years and preoperative anemia were independent predictors for postoperative allogeneic transfusion after TKA.TXA can effectively decrease the postoperative allogeneic transfusion rate and the amount of transfusion.

Objective To investigate the association and difference between patent foramen ovale(PFO), atrial septal aneurysm(ASA), atrial septal defect(ASD) in normal controls and cryptogenic ischemic stroke(CS) in youth diagnosed by double source CT.Methods A total of 168 CS patients and 180 controls matched age and gender were included in the present study.The two groups were diagnosed by double source CT and clinical materials.The incident rate of PFO, ASA, ASD, the degrees of PFO, ASD,the lengths of PFO and difference between CS in two groups were analysed.Results The incident rates of PFO, ASA, ASD were 40.6%,10.7%,6.5% and 15.6%,3.3%,2.2% in CS groups and controls respectively(P<0.001).The incident rates of PFO merged ASA and ASA merged ASD were 6.0%,3.0% and 0%,0% respectively between two groups(P<0.001).The diameters of PFO, ASD were (1.71±0.62) mm,(3.42±0.72) mm and (0.85±0.51) mm,(2.45±0.42) mm in two groups respectively(P<0.001).The lengths of PFO were (14.6±3.8) mm and (8.2±2.3) mm in two groups(P<0.001).The correlations between stroke in two groups were no difference(P>0.001).Conclusion PFO,ASA and ASD are important to CS.While PFO,ASA and ASD can accurately be diagnosed by double source CT.

Objective To evaluate the additional efficacy of local anesthetic injection (LAI) as a part of multimodal anal?gesia in patients undergoing total knee arthroplasty (TKA) with respect to pain, narcotic use, knee function and complications. Methods A multicenter randomized, controlled, double blind study was performed. A total of 101 patients undergoing unilateral TKA in two centers were randomly divided into injection group and control group. Injection group (50 cases) received local anes?thetic injection of ropivacaine (200 mg), fentanyl (1μg) and epinephrine (1∶1 000, 0.25 mg) in operation and control group (51 cas?es) did not. All patients received standardized general anesthesia and postoperative intravenous patient controlled analgesia (PCA). Preoperative baseline data, surgery?related conditions, postoperative pain (on a 0 to 10 scale), knee function, time of open?ing PCA, narcotic dosage in PCA and complications were compared respectively. Results The time of opening PCA in injection group (4-10 h, M=8 h) was longer than that in control group (2-5 h, M=4 h) (P<0.05). The 12 h, 24 h and total narcotic use of PCA in injection group (8.62±3.601 ml, 21.22±9.220 ml, 38.52±7.764 ml) was less than that in control group (18.43±9.671 ml, 35.30± 11.414 ml, 55.52±12.405 ml) (P<0.05). At post anesthesia care unit the mean VAS in injection group (2.40±1.927) was lower than that in control group (3.06 ± 2.073) (P<0.05). There was no difference in mean VAS at other time points, knee function, length of stay between two groups (P>0.05). Conclusion LIA in TKA can relieve pain early after TKA, prolong the time of opening PCA and reduce narcotic use compared with patients without it. It is simple and safe to use.

Objective To observe the effects of tumor necrosis factor-or (TNF-α) and platelet derived growth factor (PDGF) on vascular neointimal hyperplasia on matrix metalloproteinase 9/2 gene knockout (MMP9/2-/-) mice and explore related mechanisms.Methods Mice of control group,MMP9-/-group,MMP2-/-group and MMP9/2-/-group were studied.Femoral artery was injured by transluminal wire,the mRNA expression levels of TNF-o and PDGF on femoral artery were detected by RT-PCR;the protein expression of MMP9 and MMP2 were assessed by Western blot on day 0,1,3,7,14and 28 post injury.Mice in control group received TNF-α(5 ng/ml,0.10 ml),TNF-α(0.05 ml) + MMP inhibitor SB-3CT (0.50 ng/ml,0.05 ml) injection,or PDGF-bb (10 ng/ml,0.10 ml) and PDGF-bb (0.05 ml) + SB-3CT(0.05 ml) injection around injured artery,intimal hyperplasia at 2 and 4 weeks after injury was observed.Intimal hyperplasia at 2 and 4 weeks after injury was also observed in MMP9/2-/-mice.TNF-α(5 ng/ml,0.10 ml) was injected to MMP2-/-mice,PDGF-bb (0.1 ml) was injected to MMP9-/-mice around injured artery,intimal hyperplasia at 2 and 4 weeks after injury was observed.The degree of neointimal hyperplasia were observed by the Elastica-van Gieson staining and the area of neointima and media of the arteries were measured by SigmaPlot and intima ratio was calculated.Vascular smooth muscle cell (VSMC) mediums of MMP9-/-and MMP2-/-mice were stimulated by TNF-α and PDGF-bb,respectively,and migration assay,and proliferation assay were performed,relative migration and proliferation cells numbers were counted.Results (1) mRNA expression of TNF-o (235.33 ± 23.68) and PDGF-bb (3.30 ±0.56) in femoral arteries peaked at 1 day after injury,while MMP9 or MMP2 protein expression peaked at 7 or 28 days after injury.(2) In control mice,TNF-α intervention significantly enhanced intimal hyperplasia at 2 weeks after injury (2.21 ±0.05 vs.1.55 ±0.03 in blank control group,P ＜ 0.05),while PDGF-bb intervention significantly enhanced intimal hyperplasia at 4 weeks after injury (2.60 ± 0.07 vs.1.89 ± 0.04,P =0.03).(3) Intima hyperplasia was significantly higher in control group than in MMP9/2-/-group at 2 weeks (1.63 ± 0.05 vs.0.46 ± 0.01,P =0.008) and 4 weeks (2.24 ±0.06 vs.0.51 ±0.01) after injury(P =0.005).(4) TNF-o intervention stimulated intimal hyperplasia in MMP2-/-mice (intimal ratio at 2 weeks after injury:1.73 ± 0.05 vs.1.23 ± 0.03,P =0.02) and PDGF-bb intervention stimulated intimal hyperplasia in MMP9-/-mice(intimal ratio at 4 weeks after injury:2.32 ± 0.06 vs.1.35 ± 0.03,P =0.03).(5) Reduced VSMC migration was evidenced in MMP9-/-mice post TNF-α stimulation (1.45 ±0.03 vs.2.16 ±0.04 in control group,P =0.03),while reduced VSMC proliferation post PDGF was seen in MMP2-/-group (1.15 ± 0.02 vs.1.82 ± 0.04 in control group,P =0.03).Conclusions TNF-o induced MMP9 activation plays a major role on promoting VSMC migration at the first 2 weeks after vascular injury,while PDGF induced MMP2 activation plays a crucial role on VSMC proliferation on the following 2 weeks after vascular injury in this mice model.Thus,the axis of TNF-α-MMP9-VSMC migration axis and PDGF-MMP2-VSMC proliferation axis are the two major working mechanisms responsible for intimal hyperplasia post vascular injury.

Objective:To investigate the clinical characteristics of psoriasis and status quo of medical care for patients in county areas of China.Methods:This study was a cross-sectional investigation. Based on the “Qianxian Wuyin” Project (a national project for upgrating ability for psoriasis care at county level), an online questionnaire survey was conducted in the dermatology departments of 459 county hospitals in 404 pilot administrative counties across China from February to June 2023. The questionnaire included demographic information of patients (gender, ethnicity, age, place of residence, education, marital status), and clinical characteristics of psoriasis (disease course, type, comorbidities, body surface area (BSA) and previous treatment. The Dermatology Life Quality Index (DLQI) and Psoriasis Area and Severity Index (PASI) were applied for assessing the quality of life and disease severity, and completed by patients or guardian and doctors, respectively.Results:A total of 16 935 patients completed the questionnaire. The age of patients was 1-102(44.17±11.58)years, and 71.0% (12 036/16 935) were 30-59 years old. The ratio of male to female was 2.21∶1; 24.3%(4 117/16 935) of patients had high school education; there were 9 940 patients(58.7%) with previous or current smoking and/or alcohol use; 42.8%(7 218/16 855) of patients had a disease course of 1-5 years. There were 15 630 patients(92.3%) with DLQI≥10, 8 346 patients(49.7%) with PASI≥10, 15 017 patients(89.2%) with BSA≥10%. The plaque type was the most common disease type ( n=14 965, 88.7%), and spotting type ranked the second ( n=1 141, 6.8%). The most common initial site was the trunk ( n=12 309, 72.9%). Among the comorbidities, hypertension was the most common one ( n=1 681, 10.0%). There were 7 650 reports of treatment response to conventional topical drug therapy and 3 112 reports of treatment response to systemic drug therapy, with 6 269 (81.9%) and 2 493 (80.1%) reporting poor or no response, respectively. Conclusions:The survey shows that in the county areas of China, the majority of psoriasis patients are severe patients with short course of disease, plaque type is the most common type, and hypertension is the most common comorbidity; and the conventional treatment is less effective for most patients.