Objective To investigate the effects of butylphthalide injection on serum levels of inflammatory factors and oxidative stress products in patients with acute cerebral infarction. Methods A total of 120 patients with acute cerebral infarction were selected in our hospital from March 2014 to September 2016. Patients were divided into conventional treatment group (n=56) and butylphthalide group (n=64) according the therapeutic methods. The conventional treatment group was treated with improving microcirculation and resisting blood platelet aggregation, while the butylphthalide group was injected with butylphthalide (100 mL intravenous drip, twice a day) on the basis of conventional treatment. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-17, IL-23 and methane dicarboxylic aldehyde (MDA) were detected before the treatment and 7 days,14 days after the treatment. The clinical efficacy was also accessed. Results The TNF-α, IL-6, IL-17, IL-23 and MDA levels were significantly decreased 7 days and 14 days after the treatment than those before the treatment in two groups of patients (P<0.05). And the above indexes were also decreased in butylphthalide group than those of conventional treatment group (P < 0.05). No obvious adverse reactions were observed in two groups. Conclusion Butylphthalide can improve the clinical symptoms through the inhibition of inflammation and oxidative stress in patients with acute cerebral infarction.

Objective To analyze the expressions of thymidine kinase 1 (TK1) and nuclear-associated antigen Ki-67 in triple negative breast cancer (TNBC) and their clinical significances.Methods One hundred and twenty tumor tissue sections of patients with breast cancer who were performed breast conservation treatment or modified mastectomy in the Second Affiliated Hospital of Qingdao University Medical College from June 2009 to December 2010 were collected,and there were 60 cases with TNBC and 60 cases with non-TNBC.The expressions of TK1 and Ki-67 in different breast tissues were detected by immunohistochemistry.The relationships between the expression status and clinicopathologic features were analyzed.Results The positive expression rates of TK1 in TNBC and non-TNBC were 83.33％ and 51.67％ respectively,with a significant difference (x2 =13.713,P =0.000).The positive expression rates of Ki-67 expression in TNBC and nonTNBC were 68.33％ and 31.67％ respectively,with a significant difference (x2=16.133,P =0.000).In TNBC,the expression of TK1 was related to histological staging (x2 =6.125,P =0.013),but it was not related to onset age (x2 =0.809,P =0.369),menopausal stutas (x2 =1.615,P =0.204),tumor size (x2 =0.054,P =0.816) and lymphatic metastasis (x2 =0.672,P =0.412).In TNBC,the expression of Ki-67 was related to histological staging (x2 =13.145,P =0.000) and lymphatic metastasis (x2 =6.182,P =0.013),but it was not related to menopausal stutas (x2 =1.018,P =0.313),onset age (x2 =2.377,P =0.123) and tumor size (x2 =2.401,P =0.121).The expression of TK1 was positively correlated with that of Ki-67 (r =0.369,P =0.023).The results of survival analysis showed that the disease-free survival rates of 5-year were 28.20％ and 66.70％ in the TK1 positive group and TK1 negative group,and the disease-free survival rates of 5-year were 24.30％ and 64.30％ in the Ki-67 positive group and Ki-67 negative group,with significant differences (x2 =4.194,P=0.041;x2 =4.540,P =0.033).Conclusion TK1 and Ki-67 are highly expressed in TNBC,and their expressions are correlated with histological staging and survival,which are expected to become prognostic indicators.

Gene knockout, an important technology in molecular biology, has been broadly applied in industrial microbial metabolic engineering. From the basic mechanism of DNA recombination, we summarized and compared in this review different gene knockout strategies. Three most hot and important approaches, including the lambda Red recombination system using the linear dsDNA as recombination substrate, the single or double crossover homologous recombination using the circular plasmid DNA as substrate, and the transposase mediated transposition recombination, were summarized in detail. Developing frontiers and application prospects of gene knockout were further discussed.
Biotechnology ; methods ; Gene Knockout Techniques ; methods ; Industrial Microbiology ; methods ; Metabolic Engineering ; methods ; Metabolic Networks and Pathways

Objective To validate the possibility of CD133 CD34 CD44 be served as biomarkers in cancer stem cell of human lung adenocarcinoma. Methods Two kinds of culturing methods were performed to generate adhesive tumor cells and floating aggregates, and the differences of expression of CD133 CD34 CD44 between 2 kinds of cultured cells were observed by immunofluorescence. Results Floating aggregates grew more slowly, kept activity for longer period than adhesive cells (72.5% vs 47.5%,P＜0.05). Floating aggregates expressed higher level of CD133, CD34 and CD44 than adhesive cells (68.97%,82.76%,93.10% vs 5.26%,15.79%,5.26%,P＜0.01). Conclusions The combination of CD133, CD34 and CD44 probably can be used as surface markers of cancer stem cells for human lung adenocarcinoma.

Objective To discuss the influence of eldepryl on the expressions of glial fibrillary acidic protein (GFAP) and cd11b in substantia nigra and striatum in the rats with Parkinson’s disease (PD),and to clarify the regulatory role of eldepryl in the gliacytes.Methods 72 SD rats were randomly divided into control group,PD model group and eldepryl group,and each group was divided randomly into 4 d and 8 d subgroups (n=12)after the success of model preparation.The PD rat models were established by injecting rotenone in subcutaneous.The number of GFAP and cd11b positive cells and the expressions of GFAP and cd11b were detected by immunohistochemistry and Western blotting method.Results The GFAP and cd11b positive cells were all in a resting state in control group, the GFAP-positive cell body was slender and irregular and had elongated protrusions;the cd1 1 b-positive cell body was small and branch-like,and it had more slender protrusions.The GFAP and cd11b positive cells were all in a active state in model group, the GFAP-positive cell body was hypertrophy, the proj ections increased thickening;the cd1 1 b-positive cell body was more bigger, the proj ections were shorter and thicker, and the number was increased.Compared with model group, the GFAP-positive cell body and protrusions were more slender, the CD11b-positive cell body was more smaller,the projections were more slender,and the number was decreased in eldepryl group.There were a small amount of expression of GFAP and cd11b positive cells in substantia nigra and striatum in the rats in control group,and there was no significant difference between 8 d group and 4 d group(P>0.05). The number of GFAP and cd11b positive cells and the protein expression levels were significantly increased in model group compared with control group(P<0.01);there was more expression in 8 d group compared with 4 d group,but there was no significant difference(P>0.05).The number of GFAP and cd11b positive cells and the protein expression levels in eldepryl group were significantly reduced compared with model group(P<0.01);there were less expression in 8 d group compared with 4 d group, and there was significant difference (P<0.05 ). Conclusion There are activation and proliferation of the gliacytes in substantia nigra and striatum in the rats with PD,and eldepryl can inhibit the activation and proliferation of gliacytes.

Objective To analyze the efficacy and safety of hypofractionated stereotactic radiotherapy ( FSRT ) combined with temozolomide ( TMZ ) for large brain metastases ( BMs ) in a prospective phaseⅡclinical trial.Methods From 2010 to 2015, a total of 33 patients were enrolled as subjects.The median Karnofsky Performance Status scores before and after treatment were 70 and 80, respectively.The major primary tumor was non-small cell lung cancer (57.6%).The brain metastasis had a diameter of≥3 cm or a volume of ≥6 cm3 .The radiation dose was 52 Gy in 13 fractions or 52.2 Gy in 15 fractions.Patients received TMZ at a dose of 75 mg/m2 per day concurrently.The radiotherapy was followed by 6 cycles of adjuvant treatment with TMZ (150 mg/m2, days 1-5, 28 days per cycle).Patients were reexamined by magnetic resonance imaging ( MRI) during the treatment.The radiation field would be shrunk if the gross target volume ( GTV) was reduced by≥20%.The treatment outcomes were evaluated by MRI at 2-3 months after treatment.Results The total numbers of tumors and GTVs were 95 and 38, respectively. Twenty-four (63%) out of the 38 GTVs had a volume larger than 10 cm3 and the median GTV was 15.3 cm3 (5.7-142.8 cm3).Twenty-two (67%) out of the 33 patients achieved field shrinking during the treatment, and the median reduction rate of GTV was 44%( 21%-88%) .The median total dose was 59.5 Gy, and 100%and 21.2%of patients completed the concurrent and adjuvant treatment with TMZ, respectively.In all patients, the overall response rate was 97.0%;the 1-year local control, intracranial progression-free
survival, and overall survival rates were 97%, 70%, and 62%, respectively;the median survival time was 15.3 months.The main adverse reactions were grade 1-2 nausea and vomiting.One patient got grade 3 liver function impairment.Conclusions FSRT combined with TMZ is a safe and effective approach for treating large BMs.More than 50%of patients can achieve field shrinking to shorten treatment duration and reduce toxicity.Clinical Trial Registry ClinicalTrials.gov,registration number:NCT02654106.

OBJECTIVE: To evaluate the detection of humen-lung-specific X protein (LUNX) gene in micrometastases of patients with non-small cell lung cancer. METHODS: The expression of LUNX gene in tumor tissue, lung and lymph nodes was detected by reverse transcriptase-polymerase chain reaction(RT-PCR) both in 43 non-small-cell lung cancer patients (the experimental group) and 15 lung benign patients (the control group). LUNX mRNA expression in clinic pathology,stage of cancer cell differentiation, clinic stage, age, sex, smoking history, and 4 lung cancer blood markers (CEA,CA125,NSE, and CYFRA211) were evaluated. RESULTS: The expression of LUNX gene was positive in the 2 groups. LUNX gene expression was positive in 33 of the 87 lymph nodes of the 43 patients in the experimental group (37.93%), and in 2 of the 26 lymph nodes in the control group (7.69%). The LUNX mRNA positive in the lymph nodes was closely related to the pathological type, cancer cell differentiation and clinic stage(r=0.660,0.500,0.460; P=0.011,0.017,0.022, all P<0.05), while not closely related to age, sex, smoking history and 4 lung cancer blood markers (CEA,CA125, NSE, and CYFRA211) (r=0.111, 0.135,0.083,0.354; P=0.739,0.714,0.773,0.125,all P>0.05). CONCLUSION The LUNX mRNA expression detected by RT-PCR is more sensitive than by traditional ways. The expression of LUNX gene mRNA in the lymph nodes is a valuable index for the detection of micrometastases in patients with non-small cell lung cancer.
Adult ; Aged ; Biomarkers, Tumor ; biosynthesis ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; Female ; Glycoproteins ; genetics ; metabolism ; Humans ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Lymph Nodes ; metabolism ; Lymphatic Metastasis ; Male ; Middle Aged ; Phosphoproteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective:To evaluate the safety and efficiency of a modified double-tract reconstruction procedure─proximal gastrectomy with piggyback interposed jejunal single-channel reconstruction (PJIR-STR) for early SiewertⅡ adenocarcinoma of esophagogastric junction (AEG).Method:Data of 8 SiewertⅡ AEG patients at Shanxi Tumor Hospital and undergoing PJIR-STR from May 2018 to Oct 2019 were retrospectively analyzed. The gastroesophageal reflux disease questionnaire (GerdQ) was used to score the patients at 3, 6, 12, and 18 months after surgery. The severity of postoperative reflux esophagitis was assessed by gastroscopy at 3, 6 months after surgery, using the Los Angeles Classification criteria.Result:All patients recovered well after surgery without serious complications. No obvious gastroesophageal reflux was observed in all patients at different periods (All of the GerdQ scores were <8 points.) The results of gastroscopy showed that 1 patient was diagnosed as grade B reflux esophagitis at 3, 6 months after surgery, which was responsive to conservative treatment, and the other 7 patients had no grade B or above reflux esophagitis.Conclusion:PJIR-STR is a feasible, safe reconstruction with excellent efficiency of dual anti-reflux for the SiewertⅡ AEG.

Objective To quantify the setup errors measured with kV cone-beam CT (CBCT) using breast board with or without a thermoplastic head mask in breast cancer patients who received whole breast and supraclavicular nodal region irradiation.Clinical target volume (CTV) to planning target volume (PTV) in 3 directions were also calculated.Methods The study included thirty patient receiving both whole breast and supraclavicular nodal region irradiation on Elekta Versa HD linear accelerators (Elekta Oncology Systems,Crawley,UK) between June 2016 and January 2018.The setup error data were retrospectively analyzed.All the patients were immobilized in the supine position on a breast board system (Carbon fibre breast board,Civco,Iowa,USA) with both arms raised.Twenty of the patients added an extra thermoplastic head mask to immobilize the neck.A CBCT scan was taken before treatment at the first 3 to 5 fractions and then once every week.Registration with the planning CT was performed and adjusted to match the target volume of the supraclavicular nodal region and the cervical vertebra by experienced medical staff,the position of the arms and the breast were also considered.For all patients the couch shifts in left-right (x),superior-inferior (y),anterior-posterior (z) were recorded.Statistical analysis included two-tails significance tests (t-Student and Manne-Whitney test for means,x2 test for variances).Population of the two groups was compared in terms of distribution of the mean shift (systematic error) and their standard deviations (random error).Van Herk's setup margin defined as MPTV =2.5Σ + 0.7δ was calculated in patients with or without mask immobilization.Results Altogether,56 images and 109 images were acquired in breast board only group and head mask group,respectively.Shifts of the breast board only group and the head mask group in x,y,z were 0.212±0.174 cm vs.0.272±0.242 cm (P=0.070);0.364±0.246 cm vs.0.242±0.171 cm (P=0.001);0.423±0.302 cm vs.0.364±0.269 cm (P=0.204).Proportion of the shift less than 5 mm in the breast board only group and the head mask group were 91.07％ vs.85.32％ (P=0.294);67.86％ vs.89.91％ (P=0.001);67.86％ vs.74.31％ (P=0.381).The CTV to PTV margin in x,y,z were 0.645 cm,0.981 cm,1.317 cm in breast board only group and 0.873 cm,0.709 cm,0.961 cm in head mask group,respectively.Setup error in the x direction was significantly correlated with BMI (P=0.001).Conclusions For the alignment and immobilization of patients who received whole breast and supraclavicular nodal region irradiation,using a breast board in combination of a thermoplastic head mask may significantly help to reduce the shift variance in superior-inferior direction compared to using breast board only.The anterior-posterior error is relatively large,other immobilization device or patient alignment methods are needed to be further explored to improve the accuracy.

Objective To retrospectively analyze and compare the clinical efficacy and safety between fractionated stereotactic radiotherapy (FSRT) combined with and without temozolomide in the treatment of large brain metastases.Methods Between 2009 and 2017,84 patients with large brain metastases (tumor size ≥ 6 cm3) were recruited and assigned into the CRT group (concurrent TMZ and FSRT,n=42) and RT group (FSRT alone,n=42).The radiation dose was 52.0 Gy in 13 fractions or 52.5 Gy in 15 fractions.Patients were reexamined by magnetic resonance imaging (MRI) during treatment.The radiation field would be shrunk if the gross target volume (GTV) was reduced.The clinical efficacy was evaluated at postoperative 2 to 3 months.The primary end-point event was local recurrence-free survival (LRFS) and the secondary end-point events included intracranial progression-free survival (IPFS),progression-free survival (PFS),overall survival (OS),brain metastasis-specific survival (BMSS) and adverse events.The survival rates were assessed with Kaplan-Meier method and log-rank test and monovariate analysis.Results The median GTV in the CRT and RT groups was 16.9 cm3 and 15.7 cm3.During the treatment,75％ of the lesions in the CRT group were reduced compared with 34％ in the RT group (P=0.000).The local control (LC) rate in the CRT and RT groups was 100％ and 98％.The median follow-up time was 16.1 months (range,2.1-105.7 months).In the CRT group,the LRFS (P=0.040),IPFS (P=0.022),PFS (P=0.045),OS (P=0.013) and BMSS (P=0.006) were significantly better than those in the RT group,respectively.In the CRT group,the incidence of grade Ⅰ-Ⅱ gastrointestinal adverse events was 33％,significantly higher compared with 26％ in the RT group (P=0.006).No grade Ⅳ-Ⅴ adverse events occurred in both groups.Conclusion Combined application of temozolomide and FSRT can further enhance the LC and survival rates and do not increase the risk of severe adverse events in patients diagnosed with large brain metastases.