Objective To investigate the usefulness of calcium and phosphorus supplementation in parenteral nutrition for extremely low-birth-weight (ELBW) infants.Methods According to the inclusion criteria,66 ELBW infants hospitalized after birth in neonatal ward of Tongji Hospital from June 2009 to December 2012 were divided into three groups with random number table:28 infants in the first group were treated with parenteral nutrition without calcium and phosphorus supplementation,21 infants in the second group were treated with parenteral nutrition with calcium supplementation only,and 17 infants in the third group was treated with parenteral nutrition with calcium and phosphorus supplementation.The blood levels of ionic calcium,blood phosphorus,and urine calcium and creatinine were determined once a week (total of 4 times).The speed of sound (SOS) was detected for shin bones by quantitative ultrasound at the date of admitting and the end of 4weeks.Results After administrations of supplementation,the blood levels of ionic calcium in the second group were significantly higher than those in the first group [on the 14th day,(1.82 ± 0.35) mmol/L vs.(1.14 ±0.47) mmol/L,t=5.800,P=0.005;onthe21stday,(1.77±0.45) mmol/Lvs.(1.07±0.43) mmol/L,t=5.492,P=0.004; on the 28th day,(1.61±0.58) mmol/Lvs.(0.92±0.44) mmol/L,t=4.556,P=0.025].The blood levels of ionic calcium in the third group were also significantly higher than those in the first group [on the 14th day,(1.55 ± 0.30) mmol/L vs.(1.14 ± 0.47) mmol/L,t =3.570,P =0.001 ; on the 21st day,(1.58 ±0.38) mmol/L vs.(1.07 ±0.43) mmol/L,t =4.151,P=0.000; on the 28th day,(1.55 ±0.35) mmol/L vs.(0.92 ±0.44) mmol/L,t =5.302,P =0.003].The blood levels of phosphorus were significantly elevated in the third group compared with those in the first group [on the 14th day,(1.86 ±0.10) mmol/L vs.(1.65 ±0.17) mmol/L,t=5.217,P=0.012; on the21st day,(1.88 ±0.14) mmol/Lvs.(1.61 ±0.13) mmol/L,t =6.442,P=0.003; on the 28th day,(1.89 ±0.15) mmol/L vs.(1.58 ±0.14) mmol/L,t =6.891,P =0.000] and the second group [on the 14th day,(1.86 ± 0.10) mmol/L vs.(1.53 ±0.15) mmol/L,t =8.100,P=0.000; on 21st day,(1.88 ±0.14) mmo/Lvs.(1.57 ±0.14) mmol/L,t =6.787,P =0.000; on the 28th day,(1.89 ± 0.15) mmol/L vs.(1.62 ± 0.18) mmol/L,t =5.043,P =0.000].The calcium-to-phosphorus ratios markedly increased in the second group compared with those in the first group (on the 14th day,0.69 ±0.18 vs.0.33 ±0.14,t =7.601,P =0.000; on the 21st day,0.66±0.16 vs.0.37 ±0.14,t =6.62,P=0.001 ; on the 28th day,0.62 ±0.15 vs.0.39 ±0.12,t =5.776,P =0.005) while declined in the third group (on the 14th day,0.14 ± 0.10 vs.0.33 ± 0.14,t =5.294,P =0.010; on the 21st day,0.13 ± 0.12 vs.0.37 ± 0.14,t =6.102,P =0.002; on the 28th day,0.12 ± 0.11 vs.0.39 ± 0.12,t =7.711,P =0.000).The third group showed significantly increased SOS values than those in the first and second groups [(381 ± 87) m/s vs.(135 ± 87) m/s,t =9.815,P =0.000;(381 ±87) m/s vs.(146 ±68) m/s,t =9.774,P=0.000].Conclusions Proper supplementation of calcium and phosphorus via parenteral nutrition can achieve increased bone mineral contents and stable blood ionic calcium and phosphorus levels,and avoid hypercalciuria in ELBW infants.PN therapy with calcium and phosphorus supplementation in ELBW infants needs further studies.

Objective To observe the effects of angiostatin on the activity of extra-cellular signal-regulated protein kinase (ERK) of retinal microvascular endothelial cells of mice. Methods Angiostatin was separated and purified by l-lysine sepharose 4B from human plasma. The primary retinal microvascular endothelial cells were divided into 4 groups: the control group, vascular endothelial growth factor (VEGF) 10 ng/ml group, angiostatin 130 ?g/ml group, and VEGF (10 ng/ml) + angiostatin (130 ?g/ml) group. The expression of ERK-1 was assayed by Western-blotting method 1, 2, 5, 10, 15, and 30 minutes after the treatment of angiostatin. Results Compared with the control group, the expression of ERK-1 reduced 1 minute after treatment, reduced markedly after 10 minutes. After 30 minutes, no differences of the expression of ERK were seen between the control group and angiostatin group. The activation of ERK-1 of retinal microvascular endothelial cells occurred after stimulated by VEGF, and at the pitch at the peak after 5 minutes. The level of ERK in VEGF group increased 210% than that in the control ( P

One hundred and forty-nine patients with moderate and severe organophosphorus poisoning were treated in Fangshan District Hospital, Beijing during July 2005 to July 2007, 77 cases of them with intravenous atropine and intramuscular pyraloxime methylchloride and 72 with intramuscular penehyclidime hydrochloride plus pyraloxime methylchloride. Time of atrepinization, number of medication injection, time of symptom disappearance, recovery time of cholinesterase, length of hospital stay in those with penehyclidime hydrochloride plus pyraloxime methylchloride were all better than those with atropine and pyraloxime methylchloride (P<0.05). Percentage of atropine poisoning, atropine dependence, organophosphorus poisoning rebound and adverse reaction in those with penehyclidime hydrochloride plus pyraloxime methylchloride were lower than those in those with atropine and pyraloxime methylchloride (P< 0.05).

Objective To express and detect the antigenicity of human laminin alpha4 LG3-4 module (hLN?4LG3-4) protein by gene engineering techniques. Methods The cDNA encoding hLN?4LG3-4 was amplified by RT-PCR from human placenta, then inserted into pMD-18T vector by T/A cloning and sequenced. Prokaryotic expression vector pET-28a-LG3-4 was constructed by recombinant DNA technique. The hLN?4LG3-4 fusion protein expressed in BL21(DE3)/pET system was identified by SDS-PAGE, purified by Ni-NTA resin, and assayed by Western blotting. Results The cDNA fragment of hLN?4LG3-4 was cloned successfully. While BL21 (DE3)/pET28a-LG3-4 bacterium was induced with IPTG, a new protein band with a relative molecular weight of 44 000 was shown on SDS-PAGE profile. hLN?4LG3-4 fusion protein of high purity (95%) was obtained and specific protein band was detected by Western blotting. Conclusion The hLN?4LG3-4 fusion protein was successfully expressed.

Objective To analyze the genetic characteristics of measles virus strains causing two outbreaks in Guizhou province from November 2014 to March 2015. Methods Throat swab samples collect-ed from measles cases in two outbreaks were inoculated into Vero/SLAM cells. Viral RNAs were extracted from positive cultures. Nucleoprotein genes were amplified by using reverse transcription-polymerase chain reaction ( RT-PCR) and the PCR products were sequenced and analyzed. Results Eleven strains of wild-type measles virus were isolated from the two measles outbreaks and all of them belonged to H1a sub-geno-type. Phylogenetic analysis showed that those strains were clustered into two distinct branches. Differences in nucleotide and amino acid genetic distances between the 11 strains of measles virus and the WHO reference strain of H1a sub-genotype (Chin9322) were 1. 1%-1. 6% and 0. 7%-3. 4%, respectively. Compared with the reference strain Chin9322 and Guizhou epidemic strains in recent years, six strains showed amino acid sequence mutations in 47 ( G to S) , 82 ( S to G) and 122 ( R to K) sites and two strains had a mutation in 98 ( P-L) site. Conclusion H1a sub-genotype measles virus was the predominant pathogen causing two measles outbreaks in Guizhou province during 2014 to 2015. Moreover, it was also a predominant sub-geno-type circulating in China and Guizhou province. Different measles virus strains of H1a sub-genotype contin-ued to be prevalent in Guizhou province. This study provided some scientific data for the control and elimina-tion of measles in Guizhou province.

Objective To evaluate the effect of caffeine citrate on oxygen metabolism in brain and intestine in premature infants with neonatal respiratory distress syndrome (NRDS).Methods Preterm infants aged 30-34 weeks with NRDS admitted in Guangzhou Women and Children's Medical Center during May 2015 and April 2016 were enrolled.They were administrated with maintainance dose of caffeine citrate at the 2nd day after birth.The oxygen metabolism in brain and intestine 1 h before,during and after the administration were recorded by near infrared spectroscopy.Results The cerebral oxygen saturation (ScO2) showed a less significant change before,during and after treatment.While a marked increase was seen in intestinal oxygen saturation (SsO2) during and after caffeine citrate administration,as well as the change of SsO2/ScO2 than before.SsO2/ScO2 was dramatical higher in infants with nasal continuous positive airway pressure (nCPAP) than in those with incubator oxygen supply and mechanical ventilation during and after caffeine citrate treatment than before.Conclusion Caffeine citrate may improve the delivery of oxygen and may increase the oxygenation in local tissues for preterm infants with NRDS,especially for infants with nCPAP.

Objective:With selenomethionine as reference,we investigated the bioavailability and toxicity of methylselenocysteine. Method:Selenium retention,glutathione peroxidase and thioredoxin reductase activities were tested to compare bioavailability. Acute toxicity and short-term selenium-tolerance were studied to compare toxicity. Results:Selenomethionine and methylselenocysteine were comparable in ability of increasing selenoenzymes. Selenomethionine caused much higher selenium retention,especially at higher levels and lower toxicity indicated by acute toxicity and short-term selenium-tolerance. Conclusion:Compared with selenomethionine,methylselenocysteine has equal efficacy on selenoenzymes with the characterization of less retention,but higher toxicity.

Objective:To detect the characteristics of bacteria in the feces of patients with rheumatoid arthritis (RA) and to further discover the relationship between intestinal flora and the status of peripheral cytokine, which might be able to provide new ideas for clinical treatment.Methods:The bacterial diversity and abundance of 111 RA patients and 100 age-and gender-matched healthy controls (HC) were detected by 16S high-throughput sequencing platform and compared. Based on the 16S rDNA high-throughput sequencing platform, the 16S rDNA V3 region in the participants' fecal specimens were analyzed and compared to screen for different bacterial groups. Alpha diversity was analyzed by the mothur software and the screening for different flora was tested by using Mann-Whitney, and the relationship between intestinal flora and peripheral cytokines were analyzed, too.Results:There was no significant difference in gender ( χ2=0.005, P=0.947) and age ( t=0.728, P=0.467) between the two groups. Patients with RA had a lower chao1 index ( Z=-2.188, P=0.029) and ACE index ( Z=-2.078, P=0.038) of species richness, and the Shannon index ( Z=-2.064, P=0.039) and Simpion index ( Z=-2.064, P=0.039) of diversity index in the feces compared with those of HC. At the genus level, the relative abundance of Bifidobacterium ( Z=-2.388, P=0.017), Lactobacillus ( Z=-2.543, P=0.011), Clostridium sensu stricto ( Z=-3.842, P<0.01), Blautia ( Z=-2.064, P=0.039) , Clostridium Ⅺ ( Z=-2.682, P<0.01), Turicibacter ( Z=-2.437, P=0.015), Phascolarctobacterium ( Z=-3.524, P<0.01), Megasphaera ( Z=-2.87, P<0.01), Veillonella ( Z=-2.472, P=0.013), Citrobacter ( Z=-3.263, P<0.01) and Escherichia/Shigella ( Z=-4.265, P<0.01) in RA were significantly higher than those of HC ( P<0.05), Butyricimonas ( Z=-3.071, P=0.002), Odorbacter ( Z=-2.257, P=0.024), Blautia ( Z=-2.064, P=0.039), Clostridium_ⅩⅣb ( Z=-2.901, P<0.01), Lachnospiracea_incertae sedis ( Z=-2.159, P=0.031), Acetivibrio ( Z=-2.995, P<0.01), Butyricicoccus ( Z=-2.162, P=0.031) and Gemmiger ( Z=-2.949, P<0.01) relative abundance were significantly decreased in RA patients ( P<0.05). LEfSe analysis showed γ-proteobacteria and Lachnospiraceaehad the most significant difference between the two groups. Further, patients with high inflammatory cytokines such as IL-17 and TNF-α hada higher relative abundance of Prevotella. Conclusion:The diversity and abundance of intestinal flora in RA patients are significantly different from those of healthy population, which is closely related to the levels of inflammatory cytokines, suggesting imbalance of intestinal flora might be involved in the occurrence and development of RA.

Objective To evaluate the detection of culture filtrate protein 10 (CFP10) and 6000 early secretory antigenic target (ESAT-6) in cerebrospinal fluid to be used in diagnosing tuberculous meningitis. Methods Dot enzyme linked immunosorbent assay ( Dot ELISA) method that was improved by applying concentrated cerebrospinal fluid was used to detect CFP10 and ESAT-6 in cerebrospinal fluid to analyze small protein antigen secreted by M. tuberculosis. Cerebrospinal fluid of 111 subjects were collected,in which 58 specimens were clinically diagnosed as tuberculous meningitis and 53 as non-tuberculous.CFP10 and ESAT-6 were detected in cerebrospinal fluid using Dot ELISA method and the results were analyzed. Results The sensitivities of detecting CFP10 and ESAT-6 antigen were 93.1% and 91.4% respectively, and the specificities were 92. 5% and 94. 3% respectively. The sensitivities and specificities are generally higher compared with the other methods of detecting M. tuberculosis or materials of M. tuberculosis by acid-fast staining or mycobacterium tuberculosis culture and polymerase chain reaction.Conclusions Using Dot ELISA method to detect CFP10 and ESAT-6 in cerebrospinal fluid to diagnose tuberculous meningitis has a high sensitivity and specificity. Our study provided the evidence of detecting the specific antigen of M. tuberculosis to be used in diagnosing tuberculosis.

BACKGROUND:Up to date,there is not an acceptable method for isolating,culture and amplifying human adipose-derived mesenchymal stem cells (hADSCs).OBJECTIVE:To explore the most effective way to obtain highly homogenous and undifferentiated hADSCs.DESIGN,TIME AND SETTING:The in vitro cytology experiment was performed at the Key Laboratory of Pathobiology,Ministry of Education,Jilin University from June to December 2008.MATERIALS:Human abdominal adipose tissue resected in the surgery was supplied by the Affiliated Hospital of Jilin University,The informed consent was obtained from patients.METHODS:Human adipose tissue was removed connective tissue and blood vessel,followed by incubation in 0.1% type I collagenase for 60 minutes at 37℃,filtrated then centrifuged.Consequently,the subnatant precipitation was cultured with LG-DMEM containing 10% fetal bovine serum,incubated at 6-well plate with density of 1×10~9/L,and placed in incubator of 5% CO_2 at 37 ℃.The cultured cells were passaged when the cells reached 80%-90% confiuency,and the 3~(rd) passage of cells were induced to osteogenic and adipogenic differentiation.MAIN OUTCOME MEASURES:Morphological characteristics of hADSCs were observed by laser scanning confocal microscope.Immunophenotypes,cell cycle and growth curve of hADSCs were detected by flow cytometry and immunofiuorescent techniques.In addition,the multiple differentiation potential of hADSCs was detected.RESULTS:hADSCs presented fibroblast-like morphological feature with a flocked array.The 3~(rd) passage of hADSCs had unique immunophenotypes and they were positive for CD73,CD44,CD166,CD105 and CD29,but negative for CD31,CD34,CD45 and HLA-DR.Cell cycle result showed that they had the typical growth characteristics of stem calls,namely,83.81% cells stayed at G_0/G_1 stage,only 16.19% cells were stayed at S+G_2/M stage;The latent phase of the primary culture cells was 2 days prior to and after incubation,followed by 3-6 days of logarithmical proliferation,reached a peak at day 6,and entering the growth platform phase with lower growth speed.The alkaline phosphatase was positive expressed at week 2 of osteogenic induction.And the positive expression of oil-O red staining could be seen at day 3 of adipogenic induction.CONCLUSION:Cells contamination can be reduced by removing connective tissue and blood vessel of adipose tissue,and 0.1% type Ⅰ collagenase for 60 minutes can effective separated stroma cell to matrix fiber,furthermore,ensure a sufficient contact between collagenase and tissures,which provide an supportive for harvesting highly homogenous hADSCs.