Objective To observe the effects of hypoxia on the expression of connective tissue growth factor(CTGF)and matrix metalloproteinase 9(MMP-9)in lung fibroblasts at different time points.Methods Fibroblasts were cultured at hypoxic condition 0,1.5,3,6,12 h respectively.CTGF mRNA and MMP-9 mRNA level were detected in each group by reverse transcription-polymerase chain reaction(RT-PCR).The concen- tration of CTGF and MMP-9 protein in fibroblasts supernatant were determined using enzyme-linked im- munosorbent assay(ELISA).Results Hypoxia stimulated fibroblasts increased the level of CTGF mRNA with- in 1.5 h,and the levels remained at a plateau up to 6 h,and then decreased by 12 h.The level of MMP-9 mRNA increased significantly within 3 h,and the levels kept the trend for increasing.ELISA revealed that the levels of both CTGF and MMP-9 protein/cell in medium conditioned by fibroblasts exposed to hypoxia were maximal at 12 h.The level of MMP in the CTGF-Ab treated groups was significantly decreased compared to the untreated groups.Conclusions These findings suggest that hypoxia stimulates fibroblasts to release CTGF as a mitogen factor,which initiates the fibrosis cascade and airway remodeling by regulating the balance of extracellular ma- trix synthesis and degradation via MMP-9 which is secreted by fibroblast cells in response to CTGF.

To study the effect of miR-490 on Coxsackievirus B3 (CVB3) replication, HeLa cells were trans- fected with miR-490 in vitro, and infected with a Renilla luciferase (RLuc)-expressing CVB3 variant (RLuc-CVB3). The activities of RLuc in these cells were measured at 8h intervals from 0 to 40 h post-infection (p.i.), and the effects of miR-490 on RLuc-CVB3 replication were observed. In a further study, HeLa cells were transfected with either miR-490 or antisense miR-490 (AMO-miR-490), and were then infected with an enhanced green fluorescence protein (EGFP)-expressing CVB3 variant (EGFP-CVB3). The replication of EGFP-CVB3 was then determined by detecting the expression of EGFP. We observed that miR-490 could significantly inhibit the expression of RLuc in infected cells at 32 h p. i. Furthermore, in HeLa cells infected with EGFP-CVB3 at 32 h p.i., EGFP expression was also significantly inhibited by the presence of mniR-490. The inhibitory effect of miR-490 on EGFP expression in EGFP-CVB3-infected cells could be reversed by tranfection with AMQ-miR-490. These results indicated that miR-490 significantly inhibits the replication and expression of QVB3.
Enterovirus B, Human ; genetics ; physiology ; Enterovirus Infections ; genetics ; metabolism ; virology ; HeLa Cells ; Humans ; MicroRNAs ; genetics ; metabolism ; Virus Replication

Objective To investigate the therapeutic effect of large dose mucosolvan combined variant flow rate continuous positive airway by nasal mask on neonatal respiratory distress syndrome(NRDS).Methods One hundred and fourteen newborns with NRDS were randomly divided into treatment group(58 cases) and control group(56 cases),on the base of same combined therapy,the cases in control group only underwent oxygen-absorbing by head set with the flow rate 4-6 L/min,and the cases in treatment group were given large dose mucosolvan(Ambroxol Hydrochloride) 30 mg/(kg?d) + 5%GS 20 mL,for two times and variant flow rate continuous positive airway by nasal mask(NCPAP),the parameter setting flow rate 6-8 L/min,FiO_2 0.4-0.6,pressure 5-8 cm H_2O.The clinical symptom and blood gas analysis after 12 and 48 hours were observed and compared the changes of pa(O_2),pa(CO_2),pa(O_2)/FiO_2 in two groups.Results The dyspnea and groan in 44 cases in the treatment group lessoned or vanished,pa(O_2) rised and pa(CO_2) lowered,the oxygenation index obviously increased,the cases with RDS grade Ⅰand gradeⅡ had better therapeutic effect,and the cases with RDS grade Ⅲ(X-ray)and Ⅳ had not manifest effect,the total effective rate was 75.8% in treatment group and 26.7% in control group.There were significant difference in therapeutic effect and oxygenation index between two groups.Conclusions Large dose mucosolvan(combi)-ning variant flow rate continuous positive airway by nasal mask can significantly improve the ventilation and oxygenation function and there are significant therapeutic effect in NRDS,especially in the NRDS grade Ⅰand gradeⅡ,the trachea cannula may be avoided and mechanical ventilation rate may be decreased if the therapeutic method can be used in earlier period.

Objective To investigate the direct inhibitory effects of 153Sm- DTPA-c (Cys-Gly-Arg-Arg-Ala-Gly-Gly-Ser-Cys) NH2 ( 153 Sm-DTPA-c (CGRRAGGSC)) on human prostate cancer PC-3 cells. Methods 153Sm-DTPA-c(CGRRAGGSC) was synthesized by the reaction of 153SmCl3 with DTPA-c(CGRRAGGSC) using indirect synthesis method. PC-3 cells in vitro culture were divided into four study groups, groug A ( the control, with PBS only), group B with 1.5 mg/L c ( CGRRAGGSC), group C with 370 kBq 153 SmCl3 and group D with 370 kBq 153Sm-DTPA-c(CGRRAGGSC). PC-3 cell growth was assayed by 3-(4, 5-dimethylthiazol-2-yl ) -2, 5-diphenyltetrazolium bromide (MTT) method. Cell cycle and apoptosis were analyzed by flow cytometry. The expression changes of interleukin 11 (IL11 ) and IL11 receptor (IL1 1 R) in PC-3 cells were examined by Western Blot. One way analysis of variance (ANOVA) and paired-t test were applied for statistic analysis. Results The labeling yield of 153Sm-DTPA-c (CGRRAGGSC) was 85% and the radiochemical purity was 95.8%. The specific activity of 153Sm-DTPA-c(CGRRAGGSC) was 1.32 × 105 MBq/μmol. Significant inhibitory effects on the growth of PC-3 cells were found in both group C and D, with a time-dependent manner. However, no obvious inhibition was found either in group A or in group B. After 48 h,significant differences of sub-G1 peak area were found among groups, (0. 98 ± 0. 18)%, (0. 35 ±0. 10)%, (4.05 ±0.28)% and (13.38 ±0. 89)% for group A, B, C and D, respectively. Furthermore,sexpression of ILl 1R in group D was significantly lower than that in group B and C with absorbance values 0. 339 ~ 0.014, 0.338 ~ 0.019, 0.226 ~ 0. 015 for group B, C and D, respectively. Absorbance values in groups B and C were not significantly different after treatment, compared with those before treatment; however, there was difference between absorbance values after and before treatment in group D ( t = 0. 405,1. 163,135.989,P>0.05 >0.05, <0.05). Conchluion 153Sm-DTPA-c(CGRRAGGSC) can directly in hibit the cell growth and expression of human prostate cancer cells PC-3.

Objective To assess the effectiveness of prevention program on iodine deficiency disorders and iodine nutritional status of residents in Guangdong Province.Methods Probability proportionate to size sampling(PPS) was employed in surveillance of iodine deficiency disorders.Thirty counties(cities,districts) were selected in Guangdong Province.In each county(city,district) one township(street) was selected; in each township (street) one primary school was selected and in each primary school 40 children aged 8-10 were chosen to examine their thyroid and to collect salt samples at their home for determination of salt iodine.Out of the 40 children,12 children were chosen to collect urine samples for determination of urinary iodine.From the primary schools chosen,40 grade 5 students were selected for intelligence quotient(IQ) test.In the nearby of the primary schools,3 townships(towns,street) were selected and in each township(town,street) 5 pregnant and 5 lactating women were selected to collect their urine samples for determination of urinary iodine.Type-B ultrasonic was used in measuring the thyroid volume.The iodine content of urine samples was measured by the method of arsenic and cerium catalysis spectrophotometry.The iodine content of salt was determined quantitatively with the titration method.IQ was tested by Chinese combined Raven's test.According to geographical location and the implementation of iodized salt,the effects of iodized salt on iodine deficiency disorders were analyzed in the plains and the Pearl River Delta Coastal region with mild iodine deficiency(iodized salt implementation region,referred to as the plains and the PRD),historical iodine deficiency areas (iodized implementation region) and the eastern and the western coastal areas of Guangdong(areas with non-iodized salt problem,referred to as the eastern and the western Guangdong).Results A total of 1200 children aged 8 to 10 were examined by type-B ultrasonic test,and goiter rate was 3.5％ (42/1200).The differences of goiter rate between the plains and the PRD,the historical iodine deficiency areas and the eastern and the western Guangdong were statistically significant (x2 =6.6,P ＜ 0.05).The goiter rate (6.1％) in the eastern and the western Guangdong was significantly higher than that of the plains and the PRD and the historical iodine deficiency areas (3.3％,2.0％,x2 =5.6,7.1,all P ＜ 0.05).A total of 1200 salt samples were examined.The median and coefficient of variation of iodine in the salt were 31.0 mg/kg and 23.2％,respectively.Coverage of iodized salt was 97.5％(1170/1200) while 96.1％(1153/1200) of consumed iodized salt was qualified.The median urinary iodine of 1200 children aged 8-10 was 186.5 μg/L,and the differences of median urinary iodine between the plains and the PRD,the historical iodine deficiency areas and the eastern and the western Guangdong were statistically significant(x2 =5.9,P ＜ 0.05).The median urinary iodine of the eastern and the western Guangdong(162.4 μg/L) was significantly lower than that of the plains and the PRD(207.5 μg/L,x2 =8.7,P ＜ 0.01).The difference of median urinary iodine between the plains and the PRD,the historical iodine deficiency areas and the eastern and the western Guangdong was statistically significant(x2 =58.9,P＜ 0.01).The median urinary iodine of the eastern and the western Guangdong(109.6 μg/L) was significantly lower than that of the historical iodine deficiency areas and the plains and the PRD(152.9,155.2 μg/L,x2 =18.3,20.6,all P ＜ 0.05).The mean IQ of the 1208 grade 5 students was 102.8 ± 14.3.The IQ of the plains and the PRD(104.3 ± 13.9) and the historical iodine deficiency areas(102.7 ± 14.3) was significantly higher than that of the eastern and the western Guangdong(100.3 ± 14.7,t =3.8,2.1,P＜ 0.01 orP＜ 0.05).Conclusions The goal of iodine deficiency disorders elimination is achieved as scheduled in Guangdong Province.The health level of general population has been improved significantly.Iodine nutrition is in the appropriate range (100-199 μg/L) in general population but low in pregnant women.The selling of non-iodized salt in the eastern and the western Guangdong Province should be followed closely.

Objective To investigate the expression and significance of Smad 7,Smurf 1 and Smurf 2 in basal cell carcinoma and squamous cell carcinoma.Methods Biopsy specimens were resected from 14 patients with basal cell carcinoma,19 patients with squamous cell carcinoma and 30 normal controls.Quanti- tative real-time PCR and immunohistochemical techniques were utilized to assess the expression of Smad 7, Smurf 1 and Smurf 2 in these specimens.Results The gray scale for staining of Smad 7,Smurf 1 and Smurf 2 was 166.61?7.11,166.08?8.71,and 166.25?8.15 respectively in basal cell carcinoma,161.66?5.52,166.84?9.27,and 169.98?9.48 respectively in squamous cell carcinoma.The expression levels of Smad 7,Smurf 1 and Smurf 2 were all significantly increased in basal cell carcinoma and squamous cell car- cinoma in comparison with normal controls.Conclusions The over-expression of Smad 7,Smurf 1 and Smurf 2 may interfere with transforming growth factor?signaling transduction pathway through several links,therefore prevent the inhibitory effect of transforming growth factor?on epidermal proliferation,and accelerate the abnormal proliferation in above epidermal tumors.

Objective To explore the change of plasma orexin A concentration and the correlation between plasma orexin A concentration and energy intake in obese children.Methods Fasting plasma orexin A concentrations,boaly mass index(BMI) and energy intake were measured in 48 obese children(obese group) and 48 healthy children(healthy control group),and these indexes were compared,the correlation between plasma orexin A concentration and BMI,energy intake were analyzed.Results 1.The plasma orexin A concentration in obese group was significantly lower than that in healthy control group(F=5.632 P=0.008).2.In obsess group,there were negative correlation between plasma orexin A concentration and BMI(r=-0.478 P=0.012),positive correlation between plasma orexin A concentration and total energy intake(r=0.503 P=0.007),fat intake(r=0.659 P=0.006) and protein intake(r=0.381 P=0.026),and there was negative correlation between plasma orexin A concentration and carbohydrate(r=-0.316 P=0.022).3.In healthy control group,there were negative correlation between plasma orexin A concentration and BMI(r=-0.491 P=0.018),positive correlation between plasma orexin A concentration and total energy intake(r=0.512 P=0.009),fat intake(r=0.406 P=0.005),protein intake(r=0.313 P=0.020),and carbohydrate(r=0.432 P=0.025).Conclusions Orexin A may be involved in regulation of energy metabolism in obese children,and the interaction between plasma orexin A and energy intake might be different in different nutritional status in children.

Abstract?AlM:To evaluate the efficacy and safety of silicone oil removal with a 23G transconjunctival sutureless vitrectomy system linked disposable transfusion tube and self-made suction tip.?METHODS: The suction tip was made with a 23G infusion tube be cut from the end of the 5mm. lt was used to connect the disposable transfusion tube and 23G puncture cannula. The disposable transfusion tube which was cut from the end of the MaiFei's pipe was connected with the effusion box of the vitreous cutter. lntraocular silicone oil was proactive suction and removed through two incisions on pars plana ciliaris with the vitreous cutter suction system.?RESULTS:Only 13 cases (9. 8%) need suture puncture ports in 132 cases in the operation. Operation time was 7-28min. The average operation time was 15. 1± 6. 2min. ln early postoperative, there were 107 cases ( 81. 1%) appeared lower intraocular pressure (<11mmHg) and 2 cases appeared choroid detachment in 132 cases patients. Most of the patients recovered to normal but 2 case ( 1. 5%) high myopia macular hole retinal detachment occurred recurrent retinal detachment 1wk after operation. Silicone oil was removed cleanly in the most patients, only 4 cases (3. 0%) with a little silicone oil residue.?CONCLUSlON:The surgery that silicone oil is removed through two incisions with a 23G transconjunctival sutureless vitrectomy system linked disposable transfusion tube and self - made suction tip has the advantages of safe, effective, fast, economic, and it is worthy of popularization and application in clinical.

Abstract: Objective　To investigate the pollution of paralytic shellfish poisons (PSP) in shellfish sold in Hainan Province from 2018 to 2021. Methods　From 2018 to 2021, the content of 10 paralytic shellfish poisons including saxitoxin (STX), neosaxitoxin (neoSTX), gonyautoxins 1 (GTX1), gonyautoxins 2 (GTX2), gonyautoxins 3 (GTX3), gonyautoxins 4 (GTX4), gonyautoxins 5 (GTX5), decarbamoylsaxitoxin (dcSTX), decarbamoylgonyau toxins 2 (dcGTX2) and decarbamoylgonyau toxins 3 (dcGTX3) in 7 kinds of shellfish commonly sold in 13 cities and counties in Hainan province was analyzed. Results　The detection rate of PSP in 360 shellfish samples was 10.3%. Among them, the highest detection rate of STX was 5.83%, followed by GTX2 detection rate of 4.17%; the detection rate of neoSTX and GTX3 were both 1.67%; the detection rate of GTX1 was 1.39%. None of the five PSP, GTX4, GTX5, dcSTX, dcGTX2 and dcGTX3, were detected. Four types of PSP were detected in fanscallops, two were detected in oysters, mussels and Scapharca subcrenata, only one was detected in scallops, and no toxin contamination was detected in clams and razor clams. A single sample of fanscallops detected a maximum of 4 PSP, and a single sample of oysters, scallops, mussels and Scapharca subcrenata detected a maximum of 1 PSP. The equivalence of PSP in all samples was ND-155.6 μg/kg.The annual detection rate of PSP from high to low was: 20.0% in 2020, 15.6% in 2019, 5.3% in 2018, and 2.0% in 2021, and none of the samples tested exceeded the standard. Continuously detectable STX in 2018-2020, all PSP that could be detected in 2018 were STX. In 2019, in addition to STX detected in scallops and Scapharca subcrenata, neoSTX was also detected in oysters, mussels and Scapharca subcrenata. In 2020, PSP was only detected from scallops, and GTX2 could be detected in all positive specimens, while 5 STX, 5 GTX1 and 6 GTX3 were detected. Only GTX2 detected from scallops in 2021. STX was detected in shellfish sold in 12 cities and counties, GTX2 can be detected in 10 cities and counties, neoSTX can be detected in 5 cities and counties, GTX1 and GTX2 were detected in 4 cities and counties respectively. Shellfish sold in Wenchang and Lingshui markets can detect 5 types of PSP. Conclusion　Some types of shellfish on the market in Hainan are contaminated with some kind of PSP pollution risks, and it is necessary to strengthen the supervision of PSP in marketed shellfish.

OBJECTIVETo study the effects of kneepad on expression of Bcl-2 and p53 mRNA of chondrocyte in white rabbits with knee osteoarthritis, so as to explore and treatment mechanism of OA kneepad on apoptosis of chondrocytes of rabbits with knee osteoarthritis in molecular degree.

METHODSForty-four Japanese healthy 6-month-old rabbits (equal male and female,the weight ranging from 2 to 2.2 kg) were used to establish knee osteoarthritis models by modified Hulth method. The rabbits were randomly divided into 6 groups: normal group, model group, control group (microwave), experimental group 1 (electricity), experimental group 2 (thermal), experimental group 3 (kneepad). Ten rabbits in the normal group were breed with conventional method; 9 rabbits in the model group were breed with conventional method after model made; 9 rabbits in the control group were treated with microwave for 30 minutes, one time daily; 9 rabbits in the experimental group 1 were treated with electricity (density wave) for 30 minutes,one time daily;8 rabbits in the experimental group 2 were treated with hot (hot soft membrane) for 30 minutes, one time daily; 9 rabbits in the experiment group 3 were treated with electrothermal (OA knee pad) for 30 minutes, one time daily. All the rabbits were treated for 16 weeks and then sacrificed. The expressions of Bcl-2 and p53 mRNA of chondrocytes in knee joint were detected by using fluorescence quantitative RT-PCR method.

RESULTSAt the 16 hthek,th e OD260/OD280 value range of total RNA extracted from rabbit articular cartilage tissue in each group were all at 1.80 to 2.00,wh ich indicates high RNA purity. The p53 relative mRNA in articular cartilage cells of model group,th e control group,th e experimental group 1 ,r oup 2,gr oup 3 were overexpressed,an d Belc2 mRNA expression levels of articular cartilage cells were low expression,an d compared with the normal group there were significant differences (P < 0.01). Belc2, p53 mRNA expression in articular cartilage cells,th ere were significant differences (P < 0.01) between the control group, experimental group 1, group 2, group 3 and model group. The results between the control group, experimental group 1 ,group 2 and group 3 had significant differences (P < 0.01).

CONCLUSIONOA-kneepad can up-regulate the mRNA expression of Bcl-2 as well as down-regulate the mRNA expression of p53, thereby to inhibit the apoptosis of cartilage cells and delay the degeneration of articular cartilage changes.

Animals ; Apoptosis ; physiology ; Disease Models, Animal ; Female ; Knee Joint ; metabolism ; pathology ; Male ; Osteoarthritis, Knee ; genetics ; pathology ; therapy ; Protective Devices ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Rabbits ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; genetics