Since its establishment 30 years ago, the discipline of metabolic engineering has developed rapidly based on its deep integration with molecular biology, systems biology and synthetic biology successively, which has greatly contributed to advancing and upgrading biotechnology industry. This review firstly analyzes the current status of academic research and China's competence in the area of metabolic engineering according to the data of papers published in SCI-indexed journals in the past 30 years. Subsequently, the article summarizes the development of systems biology methods and enabling technologies of synthetic biology and their applications in metabolic engineering in the past 10 years. Finally, the major challenges and future perspectives for the development of metabolic engineering are briefly discussed.
Biotechnology ; Industry ; Metabolic Engineering ; Synthetic Biology ; Systems Biology

Malignant peripheral nerve sheath tumors(MPNST) of the bladder is a very rare malignant tumor, usually secondary to neurofibromatosis type 1 (NF-1), with a few sporadic cases. This paper reports a case of 70-year-old man with bladder MPNST who underwent transurethral resection. Adjuvant intravesical instillation with gemcitabine was given after surgery, and there was no local recurrence and distant metastasis after 12-month follow-up. This paper also made a corresponding literature review.

Objective To investigate the clinical efficacy and side reaction of brucea javanica oil ( BJO) combined with 125I and chemotherapy on stageⅢ?Ⅳpatients with non?small cell lung cancer ( NSCLC) . Methods One hundred and twenty cases on stageⅢ?Ⅳpatients with NSCLC were randomly divided into two groups,60 cases received BJO combined with 125I and chemotherapy treatment(observation group),the other 60 cases received 125I combined with chemotherapy treatment(control group). Results The objective response rate(ORR) and disease control rate (DCR) were 71. 7%,86. 7% of observation group and 66. 7%,85. 0% of control group,there were no significant difference(χ2=0. 352,0. 069;P>0. 05) . The improvement rate of KPS score in observation group was significantly superior to that in control group, the difference was significant (76. 7% vs. 55. 0%;χ2=6. 261,P<0. 05) . The incidence of myelosuppression and gastrointestinal adverse e?vents in observation group was significantly lower that in control group ( 68. 3% vs. 83. 3%,41. 7% vs. 61. 7%;χ2=3. 883,4. 805;P<0. 05) . Conclusion BJO combined with 125I and chemotherapy for treating on stageⅢ?Ⅳ patients with NSCLC can reduce the toxicity and side effects caused by chemotherapy,and significantly im?prove the clinical symptoms and quality of life of patients.

Objective To investigate the antitumor effects and the possible mechanisms of dendrit-ic cells co-cultured with cytokine-induced killer cells(DC-CIK)in combination with sorafenib on two lung adenocarcinoma cell lines,A549 cells(harboring KRAS gene mutation)and PC-9 cells(harboring EGFR gene mutation). Methods DC and CIK cells were routinely generated in vitro by stimulating PBMCs isola-ted from lung cancer patients with different cytokines and then co-cultured after a week of culturing. Flow cy-tometry analysis(FCM)was used to analyze the phenotype of DC-CIK cells after 7 days of co-culturing. The 50% inhibitory concentration(IC50 )of sorafenib against tumor cells was detected by MTT assay. The tumor cells were treated with DC-CIK cells alone or in combination with sorafenib. The proliferation of tumor cells was tested by CCK-8 kit and dynamically monitored by real-time cellular analysis(RTCA). Annexin-V/ PI staining was used to examine the apoptosis rates in each group. Real-time fluorescent quantitative PCR and FCM were respectively performed to detect the expression of natural killer group 2 member D ligands (NKG2DLs)at mRNA and protein levels after the treatment with sorafenib for 24 h. Results There was no significant difference between the IC50 of sorafenib against A549 and PC-9 cells after a 24-hour exposure(P﹥0. 05). Compared with the DC-CIK biotherapy,treating the tumor cells with DC-CIK cells in combination with sorafenib significantly inhibited the cell proliferation and increased the total apoptosis rates of tumor cells(P﹤0. 05). Moreover,the inhibition rates to tumor cell proliferation were enhanced along with the in-crease of effect-to-target ratio(E/ T). Compared with the single-factor treatment groups,the normalized cell index(NCI)in the combined treatment group was significantly decreased. Blocking NKG2D could abate the inhibitory effect of DC-CIK cells on tumor cell proliferation(P﹤0. 05). The expression of NKG2DLs(inclu-ding ULBP1,UBLP2 and ULBP3)on tumor cells at mRNA and protein levels were increased to different ex-tent after treating with 5 μmol/ L of sorafenib for 24 h. Conclusion There was no significant different be-tween the inhibitory effects of sorafenib on the proliferation of lung adenocarcinoma cancer cells harboring KRAS or EGFR gene mutation. The antitumor effects of DC-CIK cells combined with sorafenib on lung ade-nocarcinoma cells might be induced by regulating the NKG2D-NKG2DLs pathway and enhancing apoptosis. Moreover,the antitumor effects of the combined treatment were better than those of single-factor treatments.

Humans ; Multiple Myeloma ; Rectum

Objective To evaluate the effect of intrathecal resveratrol on activation of Ca2+/ calmodulin-dependent protein kinase Ⅱ (CaMK Ⅱ) in spinal dorsal horn neurons of rats with bone cancer pain.Methods Thirty-two adult female Sprague-Dawley rats,weighing 180-220 g,were equally randomized into 4 groups using a random number table:sham operation group (group S),bone cancer pain group (B group),and bone cancer pain + solvent control group (BD group).Walker 256 mammary gland cancer cell suspension (4× 105 cells/ml) 5 μl was injected into the medullary cavity of the right tibia in B,BR and BD groups.Normal saline 5 μl was given in group S.On 12,13 and 14 days after injection of mammary gland cancer cells,resveratrol 200 μg/10 μl was injected intrathecally once a day in group BR,and 1％ dimethyl sulfoxide 10 μl was intrathecally injected once a day in group BD.Before injection of mammary gland cancer cells (T0) and on 3,5,7,10,12 and 14 days after injection of mammary gland cancer cells (T1-6),mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured.The rats were then sacrificed and L4,5 segments of the spinal cord were removed for confirmation of the location of phosphorylated CaMK Ⅱ (p-CaMK Ⅱ) in spinal dorsal horn neurons (by immunofluorescence) and for detection of p-CaMK lⅡ expression (using Western blot).Results Compared with group S,MWT and TWL were significantly decreased at T2-6,and p-CaMK Ⅱ expression was upregulated at T6,and p-CaMK Ⅱ was mainly co-expressed with neurons in B,BR and BD groups.Compared with group B,MWT and TWL were significantly increased at T5,6,and p-CaMK Ⅱ expression was down-regulated at T6 in group BR.There was no significant difference in MWT,TWL,and p-CaMK Ⅱ expression at each time point between group B and group BD.Conclusion Resveratrol can alleviate hyperalgesia in rats with bone cancer pain and inhibited activation of CaMK Ⅱ in spinal dorsal horn neurons may be involved in the mechanism.

Objective To evaluate the role of chemokine CXCL12 in the spinal cord in the development of bone cancer pain (BCP) in rats and the relationship with microglial activation.Methods Thirty-two female Sprague-Dawley rats,weighing 180-220 g,were equally randomized into 4 groups (n =8 each) using a random number table:sham operation group (group S),BCP group (group B),BCP + CXCL12 neutralizing antibody group (group BC),and BCP + IgG control antibody group (group BI).BCP was induced by injecting Walker 256 mammary gland cancer cell suspension (4 × 105 cells/ml) 5 μl into the bone marrow of the right tibia of rats anesthetized with chloral hydrate in B,BC and BI groups,while the equal volume of normal saline was injected instead in group S.On 12,13 and 14 days after injection of mammary gland cancer cells,CXCL12 neutralizing antibody 10 μg/15 μl was intrathecally injected once a day in group BC,while IgG control antibody 10 μg/15 μl was intrathecally injected once a day in group BI.Before injection of mammary gland cancer cells (T0) and on 3,5,7,10,12 and 14 days after injection of mammary gland cancer cells (T16),paw withdrawal threshold to mechanical stimulation (PWMT) was measured.The rats were then sacrificed and L4,5 segments of the spinal cord were removed for determination of Iba-1 (pan-microglial marker) expression in spinal dorsal horn using immunofluorescence after PWMT measurement at T6.Results Compared with S group,PMWT was significantly decreased at T2-6,and Iba-1 expression was up-regulated at T6 in B,BC and BI groups (P ＜ 0.01).Compared with B group,PMWT was significantly increased at T5,6 and Iba-1 expression was down-regulated at T6 in BC group (P ＜ 0.01).Conclusion Chemokine CXCL12 in the spinal cord is involved in the development of BCP,and microglial activation is involved in the mechanism.

Objective To evaluate the role of spinal IKK2/NF-κB pathway in the maintenance of bone cancer pain (BCP) in rats.Methods Twenty-eight unmated adult female Sprague-Dawley rats,weighing 160-200 g,were randomly divided into 4 groups (n =7 each) using a random number table:sham operation group (group S),BCP group (group BP),BCP + normal saline group (group BN),and BCP + BMS345541 group (group BB).BCP was induced by injecting Walker 256 mammary gland cancer cell suspension 5 μl (4 × 105 cells/ml) into the bone marrow of the right tibia of rats anesthetized with chloral hydrate in BP,BN and BB groups,while the equal volume of normal saline was injected in group S.On 10-12 days after operation,selective IKK2 inhibitor BMS345541 (50 μg/10 μl) was intrathecally injected once a day in group BB,and the equal volume of normal saline (10μl) was given once a day in group BN.The mechanical paw withdrawal threshold (MWT) was measured before intra-tibia injection (T0),on 7 days after intra-tibia injection (T1),at 1 h before drug administration and 1,2,4,12 and 24 h after drug administration on day 10 after operation,and at 4 h after drug administration on day 12 after operation (T2-8).The rats were sacrificed after MWT was measured at Ts and L4-6 segments of the spinal cord were removed for determination of phosphorylated NF-κB (p-NF-κB) expression (using Western blot analysis).Results Compared with group S,MWT was significantly decreased at T1-2,and the expression of p-NF-κB was up-regulated in BP,BN,and BB groups.Compared with group BP,MWT was significantly increased at T4-6,and the expression of p-NF-κB in the spinal cord was down-regulated in group BB.Conclusion Spinal IKK2/NF-κB signaling pathway is involved in the maintenance of bone cancer pain in rats.

Objective To evaluate the role of chemokine receptor CXCR4 in spinal dorsal horn in the development of morphine tolerance in rats with bone cancer pain (BCP).Methods Forty female Sprague-Dawley rats,weighing 180-220 g,were equally randomized into 5 groups using a random number table:sham operation group (group S),BCP group (group B),BCP + AMD3100 (specific CXCR4 antagonist) group (group BA),BCP + morphine group (group BM),BCP + morphine + AMD3100 group (group BMA).BCP was induced by injecting Walker 256 mammary gland cancer cell suspension (4 × 105 cells/ml) 5 μl into the bone marrow of the right tibia of rats anesthetized with chloral hydrate.On 6 days after injection of mammary gland cancer cells,AMD3100 2 mg/kg was intraperitoneally injected twice a day for 9 days in BA group,and morphine 10 mg/kg was subcutaneously injected twice a day for 9 days in BM group.AMD3100 was intraperitoneally injected and morphine was subcutaneously injected as previously described at the corresponding time point in BMA group.Before injection of mammary gland cancer cells (T0) and on 4,6,8,10,12 and 14 days after injection of mammary gland cancer cells (T1-6),paw withdrawal threshold to von Frey hair mechanical stimulation (PWMT) was measured.The rats were then sacrificed and L3-5 segments of the spinal cord were removed for determination of c-fos expression in spinal dorsal horn using immunofluorescence.Results Compared with S group,PMWT was significantly decreased at T2-6 in B and BA groups and at T4-6 in BM group,and c-fos expression was up-regulated at T6 in BM group (P ＜0.01).PMWT was significantly higher at T3-5 in BM group and at T3-6 in BMA group than in group B (P ＜ 0.01).Compared with BM group,PMWT was significantly increased at T5,6 and c-fos expression was down-regulated at T6 in BMA group (P ＜ 0.01).Conclusion Chemokine receptor CXCR4 in spinal dorsal horn is involved in the development of morphine tolerance in rats with BCP and the mechanism may be related to activation of c-fos.

A total of 1512 adult inhabitants were randomly recruited in Zhaiji district of Guiyang city in September2009.The levels of triglyceride (TG),systolic blood pressure,diastolic blood pressure,and the prevalences of abdominal obesity and hypertension increased significantly in the subclinical hypothyroidism group conpared to the euthyroid group (P＜0.05).The prevalences of high TG,low high density lipoprotein-cholesterol,and metabolic syndrome (MS) in the subgroup Ⅳ were higher than the subgroup Ⅰ (P＜0.05).Correlation analysis revealed that TSH was positively related to TG (P＜0.05).Logistic regression demonstrated that TSH was a risk factor for MS.Either in the euthyroid or total subjects serum TSH levels in the MS group were significantly higher than those in non-MS group(P＜0.05).