Objective To investigate the effects of antisense phosphothioate oligodeoxynucleotides (AS-ODNs) W086 on drug-resistant gene CTX-M expression in Escherichia coli producing extended-spectrum β-lactamases(ESBLs).Methods AS-ODNs liposome was introduced into the purpose bacteria B052.The total colony forming unit(CFU) was counted.The bacteria growth curve was drawn by microplate reader.The inhibition effects of AS-ODNs on the expressions of drug-resistant gene CTX-M were observed by RT-PCR in B052.The minimal inhibitory concentration(MIC)was determined by fluid dilution method.Results significant growth inhibition of cells treated with W086 was observed as compared with those in cells in control treated bacteria.The number of B052 colonies significantly decreased in all W086 treated groups in a concentration dependent manner (P ＜ 0.05),while CFU of B052 was not influenced in simple liposome group,simple W086 group and controlled chain group.The expression of CTX-M was selectively inhibited.Conclusion Efficiently and specificly blocking expression of CTX-M mRNA,AS-ODNs reverses the multiple drug resistance of B052.It indicates that AS-ODNs provides a new viable strategy to reverse antibiotic resistance problem.

Objective To explore the value of procalcitonin (PCT) in diagnosis and treatment of neonatal ventilator associated pneumonia(VAP).Methods Sixty neonates needed invasive mechanical ventilation admitted to our NICU from Jan 2012 to Dec 2013 were divided into VAP group (30 cases) and nonVAP group (30 cases),depending on whether the neonates developed to VAP or not in 7 days.The VAP group then was randomly divided into PCT group (15 cases) and control group(15 cases) according to random number table.The levels of serum PCT,C-reactive protein(CRP) and WBC count were detected when the neonates suspected VAP before and after mechanical ventilation.Besides,the sensitivity,specificity,positive and negative predictive values for PCT,CRP and WBC were calculated in the diagnosis of VAP.Results The serum levels of PCT in VAP group had significant difference before and after mechanical ventilation (t =1.58,P =0.000),the levels of PCT were (0.37 ± 0.25) μg/L and (2.17 ± 1.46) μg/L respectively.The levels of PCT in non-VAP group had no significant difference (t =3.67,P =0.055).The levels of CRP and WBC showed no significant differences between VAP and non-VAP group before mechanical ventilation (P ＞ 0.05).Taking PCT ≥0.40 μg/L,CRP≥ 28 mg/L and WBC ≥ 10 × 109/L as the cut off value,the sensitivity of the three indicators diagnosis of VAP were 93.3％,73.3％ and 66.7％ respectively.PCT had the highest sensitivity 93.3 ％,specificity 73.3％,positive predictive 77.8％ and negative predictive 91.7％.The antibiotic duration of PCT group was (12.6 ± 5.6) d compared with (15.1 ± 9.1) d in control group (P =0.018).Conclusion Serum PCT has high sensitivity and specificity in the diagnosis of VAP,PCT combined with clinical pulmonary infection score can improve the diagnostic accuracy of VAP,early diagnosis and timely monitoring of serum PCT is helpful in VAP.PCT guidance substantially reduces antibiotic use in VAP,so that timely surveillance of PCT is necessary for patients on ventilator.

ObjectiveTo observe the effect of the combined therapy of ginkgo-dipyridimolum and mecobalamin for diabetic peripheral neuropathy (DPN).MethodsA total of 102 DPN cases were randomly divided into treatment group(n =50) and control group(n =52).The treatment group was given ginkgo-dipyridimolum and mecobalamin, and the control group was given only mecobalamin for 4 weeks.We observed the change of symptom and sign of both groups before and after treatment,and the MNCV and SNCV of median nerve,ulnar nerve and peroneal nerve were detected.ResultsIn the treatment group, the improvement of DPN symptoms (x2 = 10.090, P ＜ 0.01) and nervous conducting speed were much more obvious than that in the control group(MNCV of median nerve, ulnar nerve and peroneal nerve was(49.12 ± 3.32) m/s, (52.11 ± 3.12) m/s and(47.32 ± 3.21) m/s respectively; SNCV of median nerve,ulnar nerve and peroneal nerve was(47.13 ± 2.56) m/s, (55.12 ± 2.31) m/s, (46.78 ± 2.66) m/s respectively,all P ＜0.05).ConclusionThe combined therapy of ginkgo-dipyridimolum and mecobalamin could effectively improve the therapeutic outcome of DPN, and the treatment was safe and worthy of widespread use.

The effect of nateglinide or sequential treatment with metformin on glycemic stability in newly diagnosed type 2 diabetes was investigated. Thirty-four cases of newly diagnosed type 2 diabetes received nateglinide therapy, or sequential treatment with metformin according to fasting and postprandial blood glucose, and were classified into isolated nateglinide therapy group(n=14) and sequentially treated with metformin group(n=20). Glycemic stability, reflected by mean amplitude of glycemic excursions(MAGE) and HbA1C, was determined in all patients before and after therapy for three months. HbA1C and MAGE in two groups were all improved after treatment(P＜0.05). The therapy of nateglinide alone or combined with metformin can significantly improve glycemic stability in newly diagnosed type 2 diabetes.

Objective To review curative effect of coronary artery bypass grafting(CABG).Methods 10 patients are off-pump CABG at normal body temperature.The rest 222 were done with CPB under general anesthesia,hypothermia.Before operation cardiopulmonary function Was adjusted.During operation the function of heart,brain and renal Was carefully protected.After operation blood slucose was controlled,inflammation was prevented and pulmonary physical therapy was given to all patients.Results 226 patients discharged recovery and free of angina pectoris completely.3 patients with CABG died of low cardiac output after operation.1 patient died of effects of protamine allergy.2 patients died of multiple organ failure.Follow-up has been done in 211 patients for 2 months to 3 years,all leading normal life and working.Conclusion Careful and standardized preoperative and pefioperative treatment is one of the factors of SUCCeSSful coronary artery bypass grafting.

BACKGROUND:Adrenomedulin gene transfection can strength the anti-apoptotic ability of bone marrow mesenchymal stem cels under ischemia and hypoxia, but its mechanism is not yet clear.
OBJECTIVE:To investigate the effect of adrenomedulin on the expression of apoptosis-related proteins, Bax, Bcl-2 and Caspase-3, in bone marrow mesenchymal stem cels under hypoxia and ischemia.
METHODS:Bone marrow mesenchymal stem cels of Sprague-Dawley rats were isolated, cultured and purified, and then cultured in serum-free medium under hypoxic condition for 0, 3, 6, 9, 12 hours. Then, western blot assay was employed to detect the expression of Bax, Bcl-2 and Caspase-3 so as to determine the optimal hypoxia time that was determined at 6 hours of hypoxia. Depending on whether adrenomedulin pretreatment was done, the cels were divided into control group (with no adrenomedulin pretreatment before hypoxia and ischemia) and adrenomedulin groups with different concentrations (1, 10, 100 μg/L). Afterwards, the expression of Bax, Bcl-2 and Caspase-3 was detected by using western blot assay.
RESULTS AND CONCLUSION:(1) After cultured in serum-free medium under hypoxia for 0, 3, 6, 9, 12 hours, the expression of Bax, Bcl-2 and Caspase-3 in bone marrow mesenchymal stem cels were increased (P < 0.05);at 6 hours of hypoxia, the Bax/Bcl-2 ratio and Caspase-3 expression reached the minimum value (P < 0.05). (2) At 6 hours of hypoxia, the expression of Bax and Caspase-3 protein as wel as Bax/Bcl-2 ratio became the lowest in the 100 μg/L group compared with the 1 and 10 μg/L groups, but the expression of Bcl-2 protein reached the peak (P < 0.05). These findings indicate that adrenomedulin can reduce the expression of Bax/Bcl-2 ratio and Caspase-3 protein in bone marrow mesenchymal stem cels cultured in serum-free medium under hypoxic conditions, which is in a dose-dependent manner.

BACKGROUND:Previous studies have demonstrated that hepatocyte growth factor (HGF) gene transfection can improve the effectiveness of bone marrow mesenchymal stem cel transplantation, but the mechanism is stil unclear. OBJECTIVE:To observe the effects of HGF gene transfection on c-MET, Bax, Bcl-2, Caspase-3 of bone marrow mesenchymal stem cel s cultured under hypoxia and serum-free conditions. METHODS:(1) Bone marrow mesenchymal stem cel s were isolated and amplified in vitro by differential adhesion method. The infection efficiency of recombinant adenovirus Ad-HGF in bone marrow mesechymal stem cel s was tested by x-gal staining. (2) Bone marrow mesenchymal stem cel s were cultured under hypoxia and serum-free conditions for 0, 3, 6, 9, 12 hours. RT-PCR and western blot assays were used to evaluate the expression of Bax, Bcl-2, Caspase-3. (3) Bone marrow mesenchymal stem cel s were cultured under hypoxia and serum-free conditions for 6 hours, and RT-PCR and western blot assays were adopted to detect HGF, c-Met, Bax, Bcl-2 and Caspase-3. (4) Cel scratch test was used to detect the effect of HGF transfection on the migration of bone marrow mesenchymal stem cel s cultured under hypoxia and serum-free conditions for 6 hours. RESULTS AND CONCLUSION:(1) Transfection efficiency of bone marrow mesenchymal stem cel s was increased with multiplicity of infection in a dose-dependent manner. When the multiplicity of infection was 150, the transfection efficiency was 96.4%. (2) Expressions of Bax and Bcl-2 were gradual y increased with hypoxia time (P<0.05). The Bax/Bcl-2 ratio and Caspase-3 expression reach the minimum at 6 hours of hypoxia (P<0.05). (3) Compared with the control and Ad-LacZ groups, the expressions of HGF, c-Met, Bcl-2 increased, and the expressions of Bax and Caspase-3 decreased in the Ad-HGF group after 6 hours of culture under hypoxia and serum-free conditions (P<0.05). There was no significant difference between the control and Ad-LacZ groups. (4) The mobility of bone marrow mesenchymal stem cel s was higher in the Ad-HGF group than the control group and Ad-LacZ groups after 6 hours of culture under hypoxia (P<0.05). These findings indicate that transfection of HGF in bone marrow mesenchymal stem cel s can increase the expression of c-Met, Bcl-2 and decrease the expression of Bax, Caspase-3 under hypoxia and serum-free conditions, which also enhance the mobility of bone marrow mesenchymal stem cel s under hypoxia and serum-free conditions.

Objective To investigate the change of peripheral blooe T cell subsets,NK cells ane serum soluble interleukin-2 receptor(sIL-2R)concentration in patients with gastric cancer before ane after surgery,ane to uneerstane immune function status ane changes of perioperative gastric cancer patients. Methods One huneree ane thirty-five perioperative gastric cancer patients were selectee as our subjects who hospitalizee from May 2009 to May 2011 in Tumor Hospital of Shanxi Province,ane they were servee as treatment group,while 50 healthy subjects were selectee as controls. The number of CD3 + ,CD4 + ,CD8 + T cells,rate of CD4 + / CD8 +ane the NK cell ratio in blooe cells were eetectee by flow cytometry. ELIAS was appliee to measure serum sIL-2R concentration. Observee the above ineexes of control group at the same perioe,ane comparee the ineexes before operation of 1 e of the treatment group. Results The rate of NK cells,CD3 + T cells ane CD4 + / CD8 +ratio in patients at pre-operation were(10. 11 ± 3. 64)% ,(55. 60 ± 9. 61)% ,(30. 22 ± 6. 17)% ,1. 14 ± 0. 35,respectively,lower than that of control group(( 28. 39 ± 5. 81 )% ,( 68. 65 ± 7. 39 )% ,( 47. 87 ± 4. 85)% ,1. 82 ± 0. 24 respectively;t = - 5. 9,8. 6,8. 2,12. 7;P < 0. 01). CD8 + T positive rate of cells increasee from(27. 05 ± 7. 86)% to(34. 26 ± 6. 23)%(t = - 6. 5,P < 0. 01). At 14th eay after surgery,the cell immune function of the patients recoveree graeually,ane there were statistically significant eifferences in the above ineexes comparee with pre-operation(P < 0. 05). The eramatic changes were seen among patients with the late Gastric cancer TNM staging. Comparee with patients with stage Ⅳ,all above ineex were significant eifferent from that of patients with stageⅠ,Ⅱ(P < 0. 05),ane no significant eifferences was seen in patients with stageⅢ(P > 0. 05). The concentration of serum sIL-2R in patients with gastric cancer before operation was(575. 71 ± 34. 77)U/ L,higher than that of healthy persons((428. 26 ± 21. 77)U/ L,t = - 7. 9,P < 0. 01),ane serum sIL-2R levels in patients with stage Ⅲ,Ⅳ was lower than that of patients with stage Ⅰ,Ⅱ patients with low(P< 0. 05). Conclusion The immune function of patients with gastric carcinoma is relatee to tumor loae size ane eifferent pathological staging. The ineex of the ratio of NK cells ane T lymphocyte subsets,serum sIL-2R levels can be servee as ineicators for monitoring perioperative evaluation of prognosis of gastric cancer.

Objective To investigate the pathogen and incidence of ventilator-associated pneumonia (VAP) under different pH value of gastric juice,and to explore the endogenous pathogenesis of VAP.Methods Ninety-eight cases were admitted in our neonatal intensive care unit from Dec 2008 to Dec 2010.According to whether VAP happened,they were divided into VAP group (52 cases) and non-VAP group (46 cases).The relation between pH value of gastric juice and the species of pathogenic bacteria in stomach was analyzed.Results The incidence of VAP was 53.1％ (52/98).The pH value of gastric juice affected the species of pathogenic bacteria in stomach.When pH ≤ 4,the major species of pathogenic bacteria were Candida and Gram-positive bacillus; and pH ＞ 4,the major species were Gram-negative bacillus.The incidence of VAP was 34.5％ (10/29) when pH≤4;on the contrary,the incidence was 60.9％ (42/69) when pH ＞ 4.The incidence of VAP at the time of pH ＞ 4 was significantly higher than that of pH ≤4 ( x2 =5.71,P =0.017).Conclusion The pH value of gastric juice is an important factor of the species of pathogenic bacteria in stomach,especially for Gram-negative bacillus,including multi-drug resistant extended-spectrum β-lactamase-producing bacteria.The higher of pH value,the incidence of VAP is higher.

Objective To investigate the changes of serum myelin basic protein (MBP) and S100B orotein (S100B) in premature infants with periventricular leukomalacia (PVL) and their outcomes.Methods Seventy-eight premature infants with PVL (PVL group)and 43 normal infants (control group)who were hospitalized in our hospital from Nov 2007 to Jul 2008 were enrolled in the study.The infants were sampled for MBP and S100B levels on 1st,3rd,7th and 14th d after birth.Thirty normal infants and 69 infants with PVL were followed up every three months as they discharged until they were one year corrected age and their development quotients(DQ) were measured using Gesell development schedules.Results ( 1 ) The serum MBP levels increased on day 1 [ (7.61 ± 1.78 ) μg/L ],peak on day 3 [ ( 14.53 ± 3.12 ) μg/L],and then decreased.The serum MBP levels in infants with PVL group were significantly higher than those of control group at 1st,3rd,7th and 14th d after birth ( P ＜ 0.05 ).(2) The serum S100B levels increased on day 1,day 3 and day 7 [ (3.82 ±0.68),(4.41 ±0.91,),(5.78 ± 1.54) μg/L],peaked on day 7,and then decreased.The S100B levels of infants in PVL group were significantly higher than those of control group at 1st,3rd and 7th d after birth (P ＜0.05) ;and decreased on day 14 (P＞0.05).(3) Infants whose MBP and S100B levels increased at 7th day after birth had significantly decreased DQ than those of normal infants ( P ＜0.05 ).Conclusion The serum MBP and S100B levels in infants with PVL are correlated with the severity of central nervous system injury.If the serum S100B and MBP levels of PVL infants continues to rise more than 7 d,the DQ are lower,and the outcomes are poor.