Based on multiple painless needle insertion skills in clinic combined with own experience, the authors propose the opinion that different painless needle insertion methods need to be used according to the positions where acupoints are. The appropriate needle insertion mean must be selected in accord with the anatomical characteristic of acupoint position, namely, tapping insertion is applied in scalp part, skin-pinching up needle insertion is used in forehead and superficial nerve trunk, fingernail-pressure needle insertion in ocular region and positions where large blood vessels distribute, flying acupuncture insertion in face and finger tip, vibrating needle in occipitonuchal region, quivering insertion in abdomen, quick pricking by single hand in the four limbs and back, and chop acupuncture in palm and pelma. The insertion methods are systematically discussed in terms of preparation, acupoint position, manipulation and attention, etc, so that to provide the reference for painless needle insertion.
Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Humans ; Meridians ; Needles

Patent ductus arteriosus (PDA)is a common disease among premature neonates and the management of which has always been a challenge to the neonatologists. Early treatment has been advocated as the first-line approach, and drug induced closure is yet the most effective, inexpensive, and convenient method. During the last two decades,indomethacin was often given as conservative treatment, and its closure rate in PDA was approximately 46% ～ 89%. However,indomethacin may increase the number of undesirable or harmful effects including renal damage, intraventricular hemorrhage, necrotizing enterocolitis, and enterobrosis,etc. Recently,ibuprofen is rapidly emerging as a new promising drug for treatment of PDA in preterm infants with closure rate of 73.0% ～95.5%. It is used as standard therapy to close PDA,and it has less influence on the hemodynamics of kidneys, brain and gastrointestinal tract. The surgery is needed in patients with drug treatment failure and cardio-pulmonary function damage.

Objective To observe the serum estradiol level in newborns at different gestational age and the infants with retinopathy of prematurity (ROP), and explore the relationship between these changes and ROP. Methods A total of 178 newborns hospitalized in the Children's Hospital of Fudan University were enrolled in the study. They were divided into three groups based on the gestational age, ＜ 32 weeks group, 32-37 weeks group and term group. The three groups were further divided into non-oxygen and oxygen subgroups. Infants with birth weight less than 2000 g were evaluated for ROP and those diagnosed as ROP were under ROP group (n=6). Serum estradiol levels were measured every week from birth to discharge with radioimmunoassay. Results Serum estradiol levels of newborns quickly declined after birth. In the non-oxygen subgroups, serum estradiol levels in 32 ～ 37 weeks group were remarkably higher than those of term group in the first week [(3506±1376) pmol/L vs (1717±179) pmol/L,P＜0. 05] and the third week [(1431±92) pmol/L vs (996±102) pmol/L, P＜0. 05]; Serum estradiol levels in ＜32 weeks group was only remarkably higher than those of term group in the first week [(3173± 1369) pmol/L vs (1717± 179) pmol/L,P＜0. 05]; Serum estradiol level in the ＜32 week group was (560±355) pmol/L, lower than that in the 32-37 weeks group [( 1124 ± 128) pmol/L] (P＜0. 05). There was no difference of serum estradiol levels in the oxygen and non-oxygen subgroups of the same group. There was no significant difference of serum estradiol levels between the ROP group and ＜32 weeks group. Serum estradiol levels in ROP group was lower than that of premature group without ROP at 29 and 32-36 weeks of corrected gestational age, but no statistical difference was shown. Conclusions Very low serum estradiol level in premature infants may be related to the development of ROP, but further clinical studies are required.

Objective To estabolish an optimal animal model of oxygen-induced retinopathy(OIR) suitable for examining pathogenesis and therapeutic intervention for retinopathy of prematurity(ROP). Methods Fifty-four 7-day-old C57BL/6J mice were devided into two groups. Twenty-seven mice in hyperoxic group were exposed to 75% oxygen for 5 days and then to room air for another 5 days. Twenty-seven mice in normoxic control were exposed to room air for 10 days. The proliferative neovascular response was estimated by observing the vascular pattern in adenosine diphosphate-ase(ADPase) stained retina flat-mounts and quantitated by counting the number of new vascular cell nuclei extending into the internal limiting membrane in cross-sections. Results Angiography in ADP ase stained retina flat-mounts delineated the entire vascular pattern. Hyperoxia-induced neovascularization occurred at the junction between the vascularized and avascular retina in the mid-periphery in all mice exposed to hyperoxia. After 5 days of exposure to hyperoxia at postnatal day 12(P12),the larger central radial vessels became tortuous and constricted and central perfusion became decreased obviously. After return to room air for 2 days at P14,neovascularization was seen. This response was maximal at P17. There was a mean of 44 neovascular nuclei per cross-section extending into the vitreous in hyperoxia compared to less than 2 nuclei in the normoxia control ( P

Objective To explore the effect of comprehensive rehabilitation on patients with knee ankylosis and contracture of popliteal fossa after severe burn. Methods 35 patients with severe burn on lower limb (containing the around skin of popliteal fossa) accepted exercise therapy, hydrotherapy, occupation therapy (including pressure therapy), electrotherapy, etc. They were assessed with Functional Independence Measure (FIM), range of motion and muscle strength of knee before and after treatment. Results The scores of FIM, range of motion and muscle strength of knee improved after treatment (P<0.01). Conclusion Comprehensive rehabilitation can improve activities of daily living and function of knee after severe burn.

Objective To predict the risk of neonatal hyperbilirubinemia by transcutaneous bilirubin (TcB) nomograms and clinical risk factors.Methods Healthy term and late-preterm newborns (≥ 35 gestational weeks,and birth weight ≥ 2 000 g) born in Guizhou Maternal and Child Care Hospital between January 1,2013 and December 31,2013,were included.TcB levels were continuously recorded within 168 hours after birth.The value of hour-specific TcB nomogram combined with receiver operating characteristic (ROC)curves and Logistic regression model for predicting risk of hyperbilirubinemia was evaluated.Pearson's Chisquare test was also used for statistical analysis.Results A total of 5 250 cases were enrolled.TcB increased rapidly in the first 40 hours after birth,slowly increased between 40 to 96 hours,and reached a high level after 96 hours.Among them,the 95th percentile TcB stablized at 96 hours after birth.The 40th,75th and 95th percentile TcB peak levels were 173,217 and 248 μmol/L.Among the 5 250 neonates,there were 277 cases (5.3％) in the high-risk zone within 72 hours.The positive predictive value (PPV) was 22.02％;1 087 cases (20.7％) and 1 854 cases (35.3％) were in the medium-high risk and medium-low risk zones along with the PPV of 10.58％and 3.72％,respectively.There were 2 032 cases (38.7％) in the low-risk zone with the PPV of 1.38％.Multivariate analysis showed that the TcB high-risk zone after 72 hours was associated with gestational age,delivery mode,feeding mode and TcB level of risk zones within 72 hours.Compared to those born at ≥ 40 gestational weeks,those born at ≥ 37-＜40 gestational weeks were more likely in the TcB high-risk zone after 72 hours (OR=1.80,95％CI:1.29-2.51).The likelihood was reduced by 42％ among neonates born with cesarean section compared to those delivered vaginally in term of the TcB high-risk zone after 72 hours.Infants who received mixed feeding were less likely to be in the TcB high-risk zone after 72 hours when compared to breastfed infants (OR=0.51,95％CI:0.29-0.88).With the reduction of the high-risk zone level within 72 hours,the likelihood in the TcB high-risk zone after 72 hours was also decreased.ROC curve showed that the area under the curve (AUC) for predicting hyperbilirubinemia was 0.75 and its 95％CI was 0.72-0.78,with a sensitivity of 90.00％ and specificity of 40.00％.The AUC of a combination of predictive results obtained by the Logistic regression model with significant variables in univariate analysis and high-risk zone after 72 hours was 0.66,and its 95％CI was 0.62-0.69.AUC estimated by Logistic regression model according to the TcB levels of risk zones within 72 hours combining with clinical risk factors was 0.79,and its 95％CI was 0.76-0.82 (P＜0.01).Conclusions Hour-specific TcB nomograms of newborns in our hospital have been obtained,which facilitates the prediction and early intervention of neonatal hyperbilirubinemia.

Objective To construct eukaryotic expression plasmid pIRESneo3-pigment epithelium-derived factor (PEDF) and detect its transient expression in SP2/0 cells. Methods Specific primers were designed based on the mature peptide sequence of human PEDF cDNA in the GenBank. Human PEDF gene was cloned into the eukaryotic expression vector pIRESneo3. The PEDF DNA was transfected into SP2/0 with LipofectamineTM 2000. The recombinant human PEDF protein expressed in SP2/0 cell culture supernatant was identified by Western blot and enzyme-linked immunosorbent assay. The biological activity of the recombinant human PEDF was measured by 3-(4,5-dimethylthiazol-z-y1)-2,5-diphenytetrazolium bromide(MTT) method. Results PCR amplification, restriction enzyme digestion and DNA sequencing confirmed that the mature peptide sequence of human PEDF cDNA was successfully cloned into the eukaryotic expression vector pIRESneo3. And the plasmid was transfected into SP2/0 cells, which could secret PEDF. Western blot analysis showed that there was only one obvious band at the position of relative molecular weight of 50 000, and it is equivalent to the expected value. Enzyme-linked immunosorbent assay suggested that the content of PEDF began to rise after transfection, and peaked at 36 h [(0.92±0.04) μg/ml]. The proliferation of human umbilical vein endothelial cell line was significantly inhibited by supernatant after transfection of 36 h (P<0.05). Conclusions The eukaryotic expression plasmid pIRESneo3-PEDF had been successfully constructed and active human PEDF was transiently secreted, which made a foundation for further study of stable expression and purification of PEDF. This protein could be a potential medication for preventing and managing retinopathy of prematurity.

Background Retinal pigment epithelial (RPE) cell transplantation is a novel approach to the treatment of hereditary retinal diseases, however, human-derived RPE cell line occurs de-differentiation during in vitro cell culture.Studies showed that early abnormal activation of Akt/mammalian target of rapamycin (mTOR) signal pathway is the primary cause of RPE cell line de-differentiation, therefore, the inhibition of mTOR pathway will be helpful for the retard of de-differentiation of RPE cells.Objective This study aimed to investigate whether rapamycin can suppress the activation of mTOR pathway and promote differentiation of ARPE19 cells.Methods ARPE-19 cells were incubated in 12-well plate and divided into control group and rapamycin-treated group.DMSO or rapamycin with the final concentration of 400 nmol/L was added in the medium of the control group and the rapamycin-treated group, respectively.The cells of each group were collected 24 hours and 48 hours after cultured.The expression of zonula occludens-1 (ZO-1) in the cells was examined by immunofluorescence.The relative expression levels of RPE cell specific genes and proteins were assayed by using real-time quantitative PCR and Western blot, respectively.The detected results were compared between the two groups.Results ZO-1 was expressed in both group,but the fluorescence intensity was evidently enhanced in the rapamycin-treated group.The relative expression levels of RPE65, MERKT and LRAT mRNA in the cells increased by 25.97％ , 29.71％ and 13.00％ in the rapamycin treated group compared with the control group 24 hours after cultured (P=0.04,0.04,0.04) , and the expression levels of RPE65, LRAT, rLBP1, BEST1 , keratin18 and MERKT mRNA elevated by 174.00％ , 88.00％ , 56.18％ ,193.81％ ,10.83％ and 35.02％ in the rapamycin-treated group in comparison with the control group 48 hours after cultured (P =0.00,0.04,0.01,0.04,0.04,0.03).In addition, the expressions of p-mTOR, p-P70S6 and p-S6 protein were weaker in the rapamyein-treated group than those in the control group both 24 hours and 48 hours after cultured.Twenty four hours after cultured,the expression level of ZO-1 protein raised by 40％ in the rapamycin-treated group compared with the control group (P =0.01);while 48 hours after cultured,the expression levels of ZO-1 ,MERKT, catenin and LRAT proteins elevated by 36.00％ ,57.37％, 13.68％ and 41.07％ in the rapamycintreated group in comparison with the control group (P=0.01,0.00,0.04,0.04).Conclusions Rapamycin can suppress the activation of mTOR signaling pathway and up-regulate the expressions of RPE specific genes in ARPE19 cells.Inhibition of mTOR pathway might be an effective way for culturing RPE cells in vitro.

Objective To assess the reliability and validity of the Tree-Drawing Test in medical college students.Methods The study randomly selected 312 aged 19 to 23-year-old medical students to take part in TreeDrawing Test.In addition,a total of 275 college students were selected to receive re-test,30 days late and Pearson correlation coefficient of two tests were calculated.The three raters were invited to assess 30 trees painting score,analyzing the Kendall coefficient of concordance between the scores to verify raters' reliability; parts of students also participated in the 16PF test,SAS,SDS test,analyzing the correlation coefficient between the various test results,in order to assess the effectiveness of the Tree-Drawing Test.Results The re-test reliability in different time was 0.570-0.733 and 0.341-0.713 (P＜0.05),the raters' reliability was 0.491 ～ 0.626(P＜0.05),there are some correlations between Tree-Drawing Test and 16PF,SAS,SDS.Conclusion The Tree-Drawing Test has good reliability and validity; it can be applied to the detection of college students' psychological assessment and psychological problems.

ObjectivesTo present the current condition of non-invasive ventilation in newborns in the last ifve years in China, to describe the probable research trends of this ifeld, and to provide the possible research directions in future.Methods Using co-word analysis, the keywords “neonate” and “non-invasive ventilation” were searched in the CNKI database. The search results included 457 articles. Then the relation matrix was built by Excel 2010. Finally the visualized network was drawn by Ucinet 6.3.ResultsNasal continuous positive airway pressure (nCPAP) is primarily for the treatment of neonatal respiratory distress syndrome (NRDS) in China. Meanwhile, nasal intermittent positive pressure ventilation (nIPPV) and bilevel positive airway pressure (BiPAP) are gradually applied in clinic. The application scope of non-invasive ventilation is expanding. Besides the neonatologists, nurses are also paying close attention to non-invasive ventilation.ConclusionsThe visualized network, successfully built by Netdraw, relfects the hot topics and current condition in this ifeld.