OBJECTIVE:To observe clinical efficacy and safety of nitric oxide (NO) combined with sildenafil in the treatment of neonatal pulmonary hypertension. METHODS:66 cases of neonatal pulmonary hypertension were selected and randomly divided in-to observation group and control group,with 33 cases in each group. 2 groups of patients were given Dopamine injection 5 μg/(kg· min),ivgtt. Control group was given NO 5-20 ppm;observation group received oral or nasal feeding of Citric acid sildenafil tab-lets,0.5-1.0 mg/kg,96 h,on the basis of control group. Both group received 3 d of treatment. Clinical efficacy of 2 groups were observed as well as the level of PaO2,PaCO2,SaO2,PAP,NO,ET-1,HIF-1α,EPO and Ca2+before and after treatment. The inci-dence of ADR was compared between 2 groups. RESULTS:Total effective rate of observation group was 96.97%,which was sig-nificantly higher than that of control group(75.76%),with statistical significance(P<0.05). There was no statistical significance in the levels of PaO2,PaCO2,SaO2,PAP,NO,ET-1,HIF-1α,EPO and Ca2+between 2 groups before treatment(P>0.05);com-pared to before treatment,PaO2,SaO2,No and Ca2+ levels of 2 groups increased significantly after treatment,while PaCO2,PAP, EPO,HIF-1,ET-1 significantly lowered and the observation group was better than the control group,with statistical significance (P<0.05). No obvious ADR was found in 2 groups. CONCLUSIONS:Sildenafil combined with NO is effective in the treatment of neonatal pulmonary hypertension and can effectively improve pulmonary hypertension and blood gas indexes with good safety.

Objective To study the mRNA expression and function of TGF-?1, TGF-?RⅡ and CD105 in the lesional, non-lesional skin of psoriasis and of normal human skin. Methods The RNA of skin tissue was extracted using single-step method of RNA isolation by acid guanidinium thiocyanate. Reverse transcription polymerase chain reaction (RT-PCR) was used to measure the expression of mRNA. Results The mRNA expression of TGF-?1 and TGF-?RⅡ was lower in the epidermis of psoriatic lesion than that of non-lesional psoriatic and normal human skin(P

Objective To approach the significance of serum protein fingerprinting on the identification of different courses of acute myocardial infarction. Methods For 153 patients of acute myocardial infarction, including 45 cases of just admitted patients, 12 cases of 3h admitted patients, 22 cases of 6h admitted patients, 24 cases of 9h admitted patients, 24 cases of 12h admitted patients, 16 cases of 24h admitted patients and 10 cases of 48h admitted patients. The relative contents of serum proteins were detected by IMAC3 chip and proteinchip reader (CipherGen Inc., VS). Results On the M/Z values ranged from 4KDa to 12KDa, ten protein contents were obviously different between the different courses of acute myocardial infarction. All the patients in different courses were diagnosed correctly, the accuracy was 100%(153/153). Both sensitivity and specificity were also 100% in learning mode. Conclusion Different courses of acute myocardial infarction patients can be quickly and correctly diagnosed by this method with high sensitivity and specificity. That will be widely used in clinical application

Objective To establish a serum protein fingerprinting technique with a pattern matching algorithm to distinguish Dukes A stage from colorectal cancer of Dukes B,C,D stage. Methods Serum samples were collected from both reserved group and test group, each of the groups comprised 10 patients of colorectal cancer in Dukes A stage and 68 patients in Dukes B,C,D stage. The sera of the reserved group was combined with the surface of the IMAC3 proteinchip. Then the data of SELDI TOF MS was read and analyzed by BioMarker Wizard software and BioMarker Pattern software to get a classificatory pedigree tree, which is a standard configuration that can distinguish the sera of colorectal cancer patients of Dukes A from colorectal cancer patients of Dukes B,C,D, and the standard was confirmed by double blind test in the test group. Results At the M/Z values of 8 320 Da, 8 604Da, 8 867Da and 15 872Da, the protein contents in reserved group are obviously different between the two classes of patients. The accuracy of classification was 97 4%(76/78), corresponding sensitivity was 100%(10/10) and corresponding specificity was 97 1%(66/68). By double blind examination in test group, the corresponding accuracy was 100%(10/10), the corresponding sensitivity was 94 1%(64/68) and the corresponding specificity was 94 1%(64/68). Conclusion Colorectal cancer of Dukes A can be quickly and exactly diagnosed by this method with high sensitivity and specificity. That will be widely used in clinical application

Objective:To establish the bacterial endotoxin test method and the abnormal toxicity test method for reduced glutathi -one for injection.Methods:According to the requirements and methods in Chinese Pharmacopoeia (2015 edition, part IV), the bacte-rial endotoxin test and the abnormal toxicity test for reduced glutathione were studied .Results:The limit of bacterial endotoxin for re-duced glutathione was 0.125 EU· mg-1 , and the limit of abnormal toxicity was 1.0 g· kg-1 .Conclusion: The bacterial endotoxin test method and the abnormal toxicity test method are feasible .The abnormal toxicity should be supplemented in the quality standard for reduced glutathione , and the bacterial endotoxin test can replace the pyrogen test .

The relationship between M3 cholinergic receptor agonist (carbachol) hyperstimulation-induced pancreatic acinar cellular injury and trypsinogen activation or NF-kappaB activation in rats was studied in vitro. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, the active protease inhibitor (pefabloc), and NF-kappaB inhibitor (PDTC) in vitro. Intracellular trypsin activity was measured by using a fluorogenic substrate. The cellular injury was evaluated by measuring the leakage of LDH from pancreatic acinar cells. The results showed that as compared with control group, 10(-3) mol/L carbachol induced a significant increase of the intracellular trypsin activity and the leakage of LDH from pancreatic acinar cells. Pretreatment with 2 mmol/L pefabloc could significantly decrease the activity of trypsin and the leakage of LDH from pancreatic acinar cells (P < 0.01) following the treatment with a high concentration of carbachol (10(-3) mol/L) in vitro. The addition of 10(-2) mol/L PDTC didn't result in a significant decrease in the activity of trypsin and the leakage of LDH from pancreatic acinar cells treated with a high concentration of carbachol (10(-3) mol/L) in vitro (P > 0.05). It was concluded that intracellular trypsinogen activation is likely involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro. NF-kappaB activation may not be involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro.
Carbachol/*pharmacology ; Cholinergic Agonists/pharmacology ; NF-kappa B/*metabolism ; Pancreas/metabolism ; Pancreas/*pathology ; Rats, Wistar ; Receptor, Muscarinic M3/agonists ; Trypsinogen/*metabolism

Objective To investigate the role of tumor suppressor gene PTEN in colorectal cancer. Methods Using Northern blot,immunohistochemistry, colorectal carcinoma was examined in 47 cases. ResultsKG1 The expression of PTEN mRNA in colorectal cancer was lower than that in paired para-carcinoma tissues( P

Objective To explore the pathologic factors influencing the long-term survival of patients with relapsing and/or metastatic colorectal cancer (CRC) after radical operation for founding a basis for the clinical treatment protocols.MethodsThe clinical data were collected of 132 patients who were with relapsing and/or metastatic colorectal cancer (CRC) after radical resection,hospitalized from the beginning of 2002 to the end of 2007,and had integrated hospitalization and follow-up data.The relationship between the clinic pathologic factors and long-term survival was retrospectively analyzed.ResultsUnivariate analysis showed that the survival rate was higher in the patients with single recurrence and/or metastasis than in those with multiple recurrences and/or metastases,in the patients with relapsing and/or metastatic rectal cancer than in those with relapsing and/or metastatic colon cancer,and in patients received secondary operation than in those received non-operative treatment.Cox multivariable regression model analysis showed that the treatment modality for recurrence and/or metastasis,the number of recurrence and/or metastasis sites,the invasive depth of primary tumor,and lymph node metastasis were correlated with the long-term survival of patients.Among them,the treatment modality was the most important factor influencing the prognosis.ConclusionsThe recurrence and/or metastasis rate is higher in the patients with rectal cancer than in those with colon cancer,but the prognosis of the former is better than that of the latter.Secondary operation,which is helpful for the long-term survival,is suggested for the patients with relapsing and/or metastatic CRC.

Objective To explore the serum proteomics characteristics of early carcerization of post-hepatitic cirrhosis.MethodsThe serum protein profiles were detected of 62 patients with post-hepatitic cirrhosis and 100 patients with hepatic carcinoma in Ⅰ-Ⅱ stage (T1-2N0M0) and cirrhosis by surface-enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS) IMAC30-Cu2+ ProteinChip array (Ciphergen Biosystems Inc.,USA),the distinct proteins were analyzed and a classification tree model was established by using Biomarker Patterns software.The diagnostic efficacy of the model was blindly tested using the serum protein from both groups.ResultsThe serum protein profiles of post-hepatitic cirrhosis and hepatic carcinoma complicating with cirrhosis were analyzed and a classification tree model was established including 4 distinct proteins with different M/Z.In the training mode,the accuracy was 99.4% (161/162) on differential diagnosis to differentiate post-hepatitic cirrhosis and hepatic carcinoma complicating with cirrhosis,the sensitivity and specificity were 99.0% (99/100) and 100.0% (62/62),respectively,on the diagnosis of hepatic carcinoma complicating with cirrhosis.In the testing mode,the accuracy of differential diagnosis was 94.4% (153/162),the sensitivity and specificity were 95.0% (95/100) and 93.5% (58/62) on the diagnosis of hepatic carcinoma complicating with cirrhosis.ConclusionSELDI-TOF-MS ProteinChip technique is of convenient and rapid,and with high sensitivity and specificity on the diagnosis of early carcerization of post-hepatitic cirrhosis.

The recombinant defective adenovirus vector carrying human PTEN tumor suppressor gene was constructed by using AdEasy-1 system and its expression was detected in human breast cancer cell line MDA-MB-468. Human PTEN cDNA was cloned into adenovirus shuttle plasmid pAdTrack-CMV to generate a recombinant plasmid pAdTrack-CMV-PTEN, then homologeous recombination was carried out in the E. coli BJ5183 by contransforming linearized shuttle vector with adenovirus backbone plasmid pAdEasy-1. The newly recombined defective adenovirus vector Ad-PTEN containing green fluorescent protein (GFP) was packaged and propagated in 293 cells. After being purified by cesium chloride gradient centrifugation, the adenovirus was transfected into human breast cancer cell line MDA-MB-468 in vitro. The expression of PTEN mRNA and protein in infected human breast cancer cell line MDA-MB-468 was detected by RT-PCR and Western blot respectively. The recombinant defective adenovirus vector carrying PTEN gene was constructed successfully. The viral titer of purified adenovirus was 2. 5 X 10(10) pfu/mL, and about 70% breast cancer cells were infected with Ad-PTEN when multiplicity of infection (MOI) reached 50. The exogenous PTEN mRNA and protein were expressed in MDA-MB-468 cells infected with Ad-PTEN by RT-PCR and Western blot. The recombinant defective adenovirus vector of PTEN gene was constructed successfully using AdEasy-1 system rapidly, which paved a sound foundation for gene study of breast cancer.