ObjectiveTo establish a surface location method and acupuncture manipulation standard by dissecting the local structure of rat point Renying(ST9)and make a validation through the hypotensive effect of acupuncture.MethodA Wistar rat was sacrificed and fixed by cryogenic freezing. According the anatomical characteristics of human point Renying, cervical point Renying region was dissected layer by layer, and the common carotid artery, the internal carotid artery and the external carotid artery were bluntly separated. Location and measurement were made using vernier calipers and digital photographs were taken. Body surface location and acupuncture point depth were statistically analyzed to establish acupuncture manipulation standards. Point Renying and a non-acupoint were separately acupunctured to treat rat spontaneous hypertension. The changing tendency of blood pressure was statistically analyzed after four weeks.ResultAccording to the 95% reference value range, the surface location of rat point Renying was determined to be (8±0.3)mm below a line connecting bilateral mandibular angles and (5.5±0.4)mm lateral to the anterior midline, one on each side. Acupuncture manipulation standards were perpendicular insertion (5.5±0.4)mm and cautious lifting and thrusting to avoid injuring the artery. Acupuncture at point Renying had a marked hypotensive effect as compared with a non-acupoint (P<0.05).ConclusionThe surface location of rat point Renying is reliable and can be applied to animal experimental study.

Objective To investigate the clinical efficacy of topical corticosteroids combined with fusidic acid cream in treating infantile eczema.Methods Four hundreds cases of infantile eczema in the outpatient department of our hospital from March 2012 to January 2015 were collected and divided into observation group (220 cases) and control group (180 cases).The observation group received topical corticosteroids combined with fusidic acid cream,while the control group was treated only by topical corticosteroid.The medication time reaching to clinical cure,effect maintenance time during 30 d observation period and recurrence rate were recorded in the two groups.Results During the 30 days of observation, 50 cases of infantile eczema withdrew from the study,17 cases in the observation group and 33 cases in the control group.The average medication time in the observation group was (2.2±0.9)d,which was shorter than (3.2±1.1)d in the control group.The effect maintenance time during observation period in the observation group was (11.7±5.4) d,which was longer than (7.2±4.0)d in the control group,the difference was statistically significant(P<0.05);74.0% of recurrent cases during the observation period in the observation group manifested by mild eczema,while 57.8% in the control group manifested by mild eczema and had no need to use corticosteroid,the improvement of symptoms during recurrent period for the patients in the observation group was better than that for the patients in the control group,the difference was statistically significant (P<0.05).Conclusion Topical corticosteroids combined with fusidic acid cream for treating infantile eczema can reduce the medication time,prolong the effect maintenance time,and alleviate the recurrent symptoms.

Objective To examine mutations in the WT1 and PLCE1 gene in three Chinese families with autosomal recessive steroid-resistant nephrotic syndrome (SRNS) once mutations in NPHS2 had been excluded. Methods Peripheral blood samples were collected for genetic analysis from three probands of three Chinese families and their parents, and two probands' siblings, and 50 adult volunteers with normal urinalysis. Genomic DNA was isolated from peripheral blood leucocytes. Ten exons and exon-intron boundaries of WT1, and 31 exons and exon-intron boundaries of PLCE1 were amplified by polymerase chain reaction (PCR). Mutational analysis was performed by DNA sequencing directly and RFLP (restriction fragment length polymorphism) and/or PCR. Results No mutation in both WT1 and PLCE1 was identified in three probands from three Chinese families with autosomal recessive SRNS. However, three variants of WT1, 126C>T, ⅣS5-64A>G and 903A>G, and 13 variants of PLCE1, -134A>G, 810T>C, 960G>A, ⅣS11-28C>G, ⅣS15+26A>C, 4724G>C, ⅣS20+40C>T, ⅣS21+64G>A, ⅣS22-26T> A, 5320C>T, 5780A>G, ⅣS27+24A>G and ⅣS31 +48_49insT, were detected in three probands and some controls, indicating that all these variants were gene polymorphisms. WT1 polymorphism ⅣS5-64A>G, and PLCE1 polymorphism ⅣS22-26T>A were novel. Conclusion All the encoding exons and exon-intron boundaries of both WT1 and PLCE1 in three probands are examined, and no causative mutations in WT1 and PLCE1 axe found, suggesting that mutation in WT1 and PLCE1 genes is not a major cause of the Chinese families with autosomal recessive SRNS.

Objective To evaluate the role of autophagy in hippocampal neurons in cognitive dysfunction caused by sevoflurane anesthesia in juvenile rats.Methods One hundred four male SpragueDawley rats, aged 7 days, weighing 10-16 g, were randomly assigned into 3 groups using a random number table: control group (group C, n =8), sevoflurane anesthesia group (group S, n =48), and sevoflurane anesthesia + rapamycin group (group SR, n =48).Group C inhaled 60％ oxygen for 6 h.S and SR groups inhaled 3.6％ sevoflurane anesthesia for 6 h, and in addition, rapamycin 2 mg/kg was injected intraperitoneally at 1 h before of sevoflurane inhalation in group SR.The equal volume of phosphate buffer solution was given in C and S groups.At 1 h before sevoflurane anesthesia (T0) , immediately after the end of anesthesia (T1) , and at 12, 24 and 48 h after the end of anesthesia (T2-4) , 8 rats were randomly selected from S and SR groups to determine the expression of autophagy-related proteins microtubule-associated protein 1 light chain 3 (LC3) Ⅰ , LC3 Ⅱ and Beclin-1 in hippocampal neurons (by Western blot).The ratio of LC3 Ⅱ/LC3 Ⅰ was calculated.Morris water maze test was performed at 5 weeks after the end of anesthesia to assess the cognitive function.The escape latency, frequency of crossing the original platform, and duration of swimming spent at the target quadrant were recorded.Results Compared with the values at T0, the expression of LC3 Ⅱ and Beclin-1 was significantly down-regulated at T1-3 , and the ratio of LC3 Ⅱ/LC3 [was decreased in group S, and the expression of LC3 Ⅱ and Beclin-1 was significantly up-regulated at T1-4, and the ratio of LC3 Ⅱ/LC3 Ⅰ was increased in group SR (P＜0.05).Compared with group C, the escape latency was significantly prolonged, the frequency of crossing the original platform was reduced, and the duration of swimming spent at the target quadrant was shortened at 3-5 days in group S (P＜0.05) , and no significant change was found in the parameters mentioned above in group SR (P＞ 0.05).Compared with group S, the escape latency was significantly shortened, the frequency of crossing the original platform was increased, and the duration of swimming spent at the target quadrant was prolonged at 3-5 days, the expression of LC3 Ⅱ and Beclin-1 was up-regulated at T1-4 , and the ratio of LC3 Ⅱ/LC3Ⅰ was increased in group SR (P＜0.05).Conclusion Weakened autophagy in hippocampal neurons is involved in cognitive dysfunction caused by sevoflurane anesthesia in the juvenile rats.

Objective To study the effects of 5-azacytidine’s demethylation for P16 gene on hemangioma cell’s proliferation and apoptosis.Methods Bisulfite sequencing PCR was applied to detect P16 gene′s promoter methylation status in 5-azacytidine treated and untreated EOMA cell line.RT-PCR and western blot were used to detect the P16 gene mRNA and protein expressions.Flow cytometry was used to detect cell proliferation, cell cycle and cell apoptosis.The differences of P16 gene′s promoter methylation status,mRNA and protein expressions,cell proliferation,cell cycle and apoptosis in two groups were compared. Results The methylation rates in 1st and 13th CGs were 0%after 5-azacytidine treatment in EOMA hemangioma cell line,which were lower than in untreated cells.The mRNA and protein expressions increased after 5-azacytidine treatment,which were significantly higher than in untreated cells.The absorbance,S phase and G2/M phase and PI after 5-azacytidine treatment were lower than untreated cells,while the G0/G1 phase and apoptosis rates were higher.Conclusion The P16 gene promoter is hypermethylated in hemangioma cells with silent gene expressions.5-azacytidine could reverse P16 gene′s promoter methylation and silent gene expressions,which inhibit hemangioma cell’s proliferation and promote apoptosis.

Objective To assess the clinical value of transcatheter arterial chemoembolization (TACE) together with radiofrequency ablation (RFA) and sorafenib in treating recurrent hepatocellular carcinoma (HCC) after surgery.Methods A total of 40 patients with recurrent HCC after surgery, who were encountered at authors' hospital during the period from December 2009 to May 2014, were collected. The patients were divided into the study group (n=20) receiving TACE combined with RFA and sorafenib and the control group (n=20) receiving TACE plus RFA. Within 7-10 days after TACE, RFA was carried out. In the study group, oral sorafenib therapy (400 mg, two times everyday) started at 4 days after TACE. Withdrawal of sorafenib would be ordered if drug resistance occurred. Each patient underwent TACE combined with RFA not less than two times. Results The median survival time of the study group and the control group was 31.0 months and 24.8 months respectively, and statistically significant difference existed between the two groups (P<0.05). The one-year, 2-year and 3-year survival rates of the study group were 85%, 70% and 50%respectively, while the one-year, 2-year and 3-year survival rates of the control group were 80%, 55% and 30% respectively; the differences between the two groups were not statistically significant (P>0.05). The progression free survival (PFS) time of the study group and the control group was 6.8 months and 5.7 months respectively, the difference between the two groups was statistically significant (P<0.05). Conclusion TACE combined with RFA and sorafenib can prolong the overall survival time and the progression free survival time of patients with recurrent HCC after surgery.

Objective To study the effects of Lentinan on bleomycin A5’s inhibition effects in hemangioma-derived endothelial cell. Methods Hemangioma-derived endothelial cell line EOMA were divided into LTN group (only lentinan treatment), BLE group (only bleomycin A 5 treatment), L+B group (Lentinan and bleomycin A5 treatment) and CON group (no lentinan and bleomycin A5 treatment). Cell proliferation, cell cycle, PI and apoptosis were detected and compared among four groups. Results The differences of absorbance in LBE group was significantly higher than that in L+B group after treatment for d 1-d 7 (P<0.05), but both of them were significantly lower than in CON group and LTN group (P<0.05). The G 0/G 1 stage and apoptosis rate in L+B group was significantly higher than in BLE group(P<0.05), while it was significantly lower in S stage, G 2/M stage and PI(P<0.05). Conclusion Lentinan could significantly promote bleomycin A5’s inhibition effects on hemangioma-derived endothelial cell.

Sepsis should be defined as life-threatening organ dysfunction caused by a dys-regulated host response to infection. Continuous renal replacement therapy (CRRT) is one of the methods for the clinical treatment of sepsis. For patients undergoing CRRT, rational antimicrobial therapy is very important for the control of patient's infection. However, during CRRT, there is no clear guideline for the dose adjustment of antibiotics. In this paper, we analyzed the effect of CRRT combined with antibiotics on sepsis treatment in China and abroad, and discussed its effect on antibiotic clearance, and provided reference for clinical work.

Objective To investigate the effect of caffeine citrate in the treatment of neonatal apnea and the corresponding intervention measures. Methods A total of 88 children with apnea were enrolled in this study from December 2015 to February 2017, and were randomly divided into control group and study group, 44 cases in each group.The study group on the basis of conventional therapy plus caffeine citrate, the control group on the basis of conventional therapy plus aminophylline group, two newborns with apnea were duration of treatment should be 7 for 7 days, record the treatment effect and the incidence of adverse reactions. Results The total effective rate was 88.64％ in the study group and 72.73％ in the control group (P<0.05). The incidence of adverse reactions was 11.36％ in the study group and 40.91％ in the control group (P<0.05). Conclusion The application of caffeine citrate treatment of apnea with clinical efficacy and safety of the ideal of the newborn, in the course of treatment given targeted clinical nursing intervention is conducive to the protection of newborns with apnea of quality of life and life safety.

Objective To establish a real‐time quantitative PCR method for the detection of cytokine receptor‐like factor 2 (CRLF2) expression .Methods Specific primers amplification target gene CRLF2 and housekeeping genes ABL were designed ,the purified PCR products were performed the TA cloning .After bacterial colony PCR screening and sequencing ,then the recombinant plasmids DNA was extracted and measured by using UV spectrophotometer and converted to copies/mL concentration .Finally it was diluted for preparing the plasmid standard substance ,then the standard curve was drawn for observing the sensitivity and linear rang ,meanwhile the stability of the plasmid DNA was evaluated .This method was initially applied to detect the CRLF2 level of bone marrow mononuclear cells in 10 cases of healthy children and 10 cases of newly diagnosed acute lymphoblastic leukemia (ALL) .Results CRLF2 PCR product had a single specific melting curve;the linear detection range of the standard substance was 103 - 108 copies /ml;the plasmid standard substance by freeze‐thawing for 3 times remained stable;the CRLF2 level of clinical sample was within the linear detection range of standard substance .Conclusion The real‐time quantitative PCR method for CRLF2 established by our laboratory has good specificity ,linearity range and stability ,which can be applied to the quantitative detection of CRLF2 gene in clinical ALL children .