ObjectiveTo observe the efficacy of electroacupuncture in treating post-operative intractable hiccup.Method Sixty-seven patients with post-operative intractable hiccup were divided into a treatment group of 34 cases and a control group of 33 cases. The treatment group was intervened by electroacupunctureplus diaphragmatic training, while the control group was by Metoclopramide and Baclofen tablets. The effective rate, recovery time, recovery rate of the first treatment week, and relapse rate within 2 weeks were statistically analyzed.ResultThe total effective rate was 88.2% in the treatment group versus 63.6% in the control group; the mean recovery time was (5.75±3.14)d in the treatment group versus (6.11±3.40)d in the control group. The recovery rate of the first treatment week was 41.2% in the treatment group versus 18.2% in the control group, and the difference was statistically significant (P<0.05). The relapse rate within 2 weeks was 20.0% in the treatment group versus 76.2% in the control group, and the difference was statistically significant (P<0.01).ConclusionElectroacupuncture plus diaphragmatic training is an effective approach in treating post-operative intractable hiccup.

Objective To investigate the expression of Ptch-1 and Gli-1, hedgehog pathway-related genes in squamous cell carcinoma (SCC), and the effect of cyclopamine, a specific inhibitor of hedgehog signaling pathway, on the proliferation of a SCC cell line Tca. Methods Skin samples were resected from 42 patients with SCC and 10 normal human controls. Immunohistochemistry and in situ hybridization were employed to study the expression and distribution of Ptch-1 and Gli-1 in these specimens. Tca cells were incubated with cyclopamine (1, 2, 5, 10 μmol/mL) for 48 hours, or cyclopamine (5 μmol/mL) for 1-8 days. The same concentrations of lycopersicin served as the control treatment. Then, MTT assay was performed to detect the proliferation of Tca cells. A fraction of Tca cells were cultured in the presence of 5 μmol/mL cyclopamine for 72 hours followed by BrdU assay for the evaluation of cell growth and proliferation. Results A significant increment was shown in the expression of both Patch-1 and Gli-1 by immunohistochemistry (χ2= 5.656, 6.732, P<0.05, 0.01, respectively) and in situ hybridization (χ2=6.787, 9.600, respectively, both P<0.01) in SCC tissue compared with the control specimens. And both of them were predominantly distributed in the cytoplasm of SCC cells. As MTT assay revealed, cyclopamine notably inhibited the proliferation of Tea cells, and the effect increased with the concentration and action time of cyclopamine. Further more, the percentage of BrdU-positive cells was 26% in cyclopamine-treated Tca cells, significantly higher than that in the blank control cells (77%) and lycopersicin-treated cellls (72%). Conclusions Hedgehog signaling pathway is activated in the lesions of SCC, and inhibition of the pathway may facilitate the treatment of SCC.

Objective: To observe the clinical effect of puncturing Back-Shu points in treating chronic fatigue syndrome. Methods: Twenty-two subjects were recruited and treated by puncturing corresponding Back-Shu points based on syndrome differentiation. The short-form General Health Survey (MOSSF GHS) and the Chalder Questionnaires for Fatigue were adopted for evaluating the therapeutic effects. Results: Of the 22 patients, 4 cases showed a marked effect, 11 got effect, 7 got failure, and the total effective rate was 68.2%. Conclusion: Puncturing Back-Shu points is effective in relieving the symptoms of chronic fatigue syndrome and enhancing the patients' health standard.

Objective To explore the value of neutiophil gelatinase-associated lipocalin ( NGAL) taken from blood and urine samples in early diagnosis of acute kidney injury ( AKI) after heart valves replacement surgeries. Meth-ods A total of 56 patients received heart valves replacement surgeries were selected prospectively in this study. NGAL from blood and urine samples and serum creatinine ( Scr) were tested among them at different moments. AKI and non-AKI groups were divided based on Scr levels and the value of NGAL taken from blood and urine sam-ples was estimated in early diagnosis of AKI with receiver operating characteristic curve ( ROC) . Results 16 AKIs were observed among all of them. The peak value of Scr in AKI group was shown between 12 and 24 hours after surgeries, while blood-NGAL was seen high significantly (P<0.05) since 2 hours after surgeries, peak value at 4 hours, for urine-NGAL, peak value was seen at 2 hours. The area of ROC of blood-NGAL 4 hours and urine-NGAL 2 hours after surgeries for AKI diagnosis were 0.891 and 0.934, respectively. The better sensitivity and specificity were shown in both threshold set as 50 μg/L and 110 μg/L. Conclusion Blood-NGAL and urine-NGAL can be used as early diagnostic markers of AKI after heart valves replacement surgeries, whose change is significantly earli-er than that of Scr.

Objective To study to the effect of 1064-nm Q-switched Nd:YAG laser irradiation on the melanogenesis in a human epidermal melanocyte line PIG. Methods Cultured PIG cells were irradiated with 1064-nm Q-switched Nd:YAG laser (Medlite C6) at different energy densities for 10 times. After additional culture for various durations, cell viability was detected by MT assay, tyrosinase activity by dopa oxidation assay, mRNA and protein expressions of tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2 by real-time quantitative fluorescent RT-PCR and Westen blotting respectively, Results The irradiation with Q-switched Nd:YAG laser at energy densities from 1 to 3 J/cm2 had no obvious effect on the viability of PIG cells. After irradiation with Nd:YAG laser at 1 J/cm2, PIG cells showed a significant increase in the tyrosinase activity,mRNA expressions of tyrosinase and TRP-1 compared with unirradiated cells (0.563 ± 0.014 vs 0.501 ±0.019, 1.40±0.11 vs 1.0, 1.28 ± 0.03 vs 1.0, all P＜ 0.05), but both the mRNA (0.91 ± 0.17 vs 1.0, P＞0.05) and protein expressions of TRP-2 experienced no significant changes before and after the irradiation.However, a significant decrease was noted in PIG cells irradiated with Nd:YAG laser at 3 J/cm2 in tyrosinase activity, mRNA and protein expressions of tyrosinase (0.70 ± 0.02 vs 1.0, 0.64 ± 0.05 vs 1.0, both P ＜ 0.05),TRP-1 (0.73±0.04 vs l.0, 0.86±0.17 vs l.0, both P＜0.05) andTRP-2 (0.68±0.04 vs l.0,0.69±0.11vs 1.0, both P ＜0.05) in comparison with unirradiated PIG cells. Conclusions The 1064-nm Q-switched Nd:YAG laser irradiation may affect the melanogenesis in PIG cells. With no influence on cell viability, the 1064-nm Q-switched Nd:YAG laser at 1 J/cm2 could enhance melanogenesis, while that at 3 J/cm2 could suppress melanogenesis, in PIG cells.

Objective To investigate the in vitro effects of KAAD-cyclopamine, a specific inhibitor of hedgehog signaling pathway, on the growth and apoptosis of human squamous cell carcinoma cell line A431. Methods A431 cells were cultured and treated with KAAD-cyclopamine(0.5, 1, 2, 5 μmol/L).Then, MTT assay was used to detect the proliferation of A431 cells, and light microscopy to observe cell morphology at different time points with a 24-hour interval. Flow cytometry was used to assess cell cycle,and annexin-V/propidium iodide double staining to evaluate the apoptosis in these cells after 48 hours of treatment with KAAD-cyclopamine. Results KAAD-cyclopamine of 0.5, 1, 2 and 5 μmol/L inhibited the proliferation of A431 cells by (7.0±2.3)%, (20.6±2.8)%, (48.3±3.4)% and (61.6±3.3)%, respectively (F = 49.92, P＜0.01 ). Furthermore, in the presence of KAAD-cyclopamine of 5 μmol/L, on day 1, 2, 3, 4,and 5 the proliferation of A431 cells was suppressed by (18.5±2.6)%, (56.1±3.7)%, (65.4±2.8)%,(71.2±1.9)% and (75.9±3.0)%, respectively, the difference was significant among these time points(F =16.32, P＜0.01 ). Statistical analysis showed that KAAD-cyclopamine downregulated the growth of A431 cells in a dose-and time-dependent manner (r = 0.91, 0.86, P＜0.01 and 0.05, respectively). Light microscopy revealed typical morphological changes of cell damage in A431 cells. KAAD-cyclopamine in creased the percentage of cells in G1 phase from (51.8±2.9)% to(76.2±1.8)% (F = 26.34, P＜0.01 ), the proportion of hypoploid cells from (1.7±0.3 )% to (8.7±0.2)% (F = 6.32, P＜0.05 ), which suggested that KAAD-cyclopamine could arrest A431 cells in G1 phase of the cell cycle. The apoptosis ratio in KAAD-cyclopamine-treated cells was significantly higher than that in the untreated control [ (46.2±2.8)% vs (18.5±3.1 )%, F = 32.01, P＜0.01 ]. Tomatidine treatment did not affect the proliferation or apoptosis of A431 cells (both P＞0.05).Conclusion KAAD-cyclopamine can markedly suppress the proliferation and induce apoptosis of A431 cells.

Objective To evaluate the effect of Rab23 on the proliferation of a squamous cell carcinoma cell line Sa3,and to investigate its mechanisms.Methods Cultured Sa3 cells were classified into four groups:normal control group transfected with green fluorescent protein (GFP),Rab23-overexpressing group transfected with a GFP-labelled Rab23-overexpressing plasmid,Rab23-silencing group transfected with a plasmid carrying a Rab23-targeting shRNA,empty vector group transfected with an empty vector.After additional culture for different durations,plate colony formation assay and flow cytometry were performed to evaluate the proliferative activity of Sa3 cells,and Western blot was conducted to detect the expression of Erl/phosphorylated-Erk in Sa3 cells.Statistical analysis was carried out by t test,one-way analysis of variance and Bonferroni's multiple comparison test.Results Stable Sa3 cell lines with overexpression or silencing of Rab23 were established by plasmid construction and lentivirus-mediated transfection.The plate colony formation assay showed that the colony formation rate was significantly lower in the Rab23-overexpressing group than in the normal control group (2.3％ ± 0.2％ vs.3.6％ ± 0.3％,P ＜ 0.05),but higher in the Rab23-silencing group than in the empty vector group (4.1％ ± 0.2％ vs.1.8％ ± 0.03％,P ＜ 0.01).Rab23 overexpression induced G1 phase arrest in Sa3 cells.The proliferation index was significantly decreased in the Rab23-overexpressing group compared with the normal control group (0.581 ± 0.035 vs.0.698 ± 0.018,P ＜ 0.05),but increased in the Rab23-silencing group compared with the empty vector group (0.567 ± 0.015 vs.0.444 ± 0.014,P ＜ 0.01).As Western blot showed,there were no significant changes in the expression of Erk in the Rab23-silencing or-overexpressing group compared with the normal control group,whereas the expression of p-Erk was attenuated in the Rab23-overexpressing group compared with the normal control group,but enhanced in the Rab23-silencing group compared with the empty vector group.Conclusions Rab23 could inhibit the proliferation of Sa3 cells,which may be associated with the Erk pathway.

Objective To evaluate the effect of leptin on K17 expression in HaCaT human keratinocytes.Methods Some cultured HaCaT cells were treated with leptin (100 ng/ml) or remained untreated for 24 hours followed by the quantification of K17 mRNA expression by real-time PCR and detection of K17 protein expression by Western blot and immunofluorescence staining.To investigate the action mechanism of leptin,some cultured HaCaT cells were divided into several groups to be treated with leptin (100 ng/ml) alone,Piceatannol (an inhibitor of the STAT3 pathway) + leptin (100 ng/ml),PD-98059 (an inhibitor of the Erk1/2 pathway) + leptin (100 ng/ml),respectively for 24 hours,with the cells receiving no treatment as the negative control.Subsequently,the mRNA and protein expressions of K17 were measured by the above methods.Statistical analysis was done by the two-sample ttest.Results The mRNA expression of K17 was significantly higher in HaCaT cells treated with leptin alone than in those remaining untreated (3.086 7 ± 0.186 1 vs.1.000 0 ± 0.000 0,P ＜ 0.01),but significantly downregulated in HaCaT cells treated with Piceatannol + leptin and those with PD-98059 + leptin compared with those treated leptin alone (0.674 1 ± 0.060 0 and 0.855 0 ± 0.390 3 vs.2.242 7 ± 0.188 7,both P ＜ 0.01).The results of Western blot and immunofluorescence staining were in agreement with those of real-time PCR.Conclusions Leptin can induce K17 expression in HaCaT cells,likely by activating the STAT3 and Erk1/2 signaling pathways.

Objective To explore the early diagnosis value of C‐reactive protein (CRP) and procalcitonin (PCT ) for bacterial in‐fectious pneumonia of children .Methods A total of 101 children with bacterial infectious pneumonia ,64 children with non‐bacterial infectious pneumonia and 73 children with non‐infectious disease were selected in bacterial infectious pneumonia group ,non‐bacterial infectious pneumonia group and non‐infectious disease group respectively .Serum PCT and CRP levels were measured before treat‐ment in the three groups .With sputum culture results as gold standard for the diagnosis of bacterial infectious pneumonia ,the sensi‐tivity ,specificity ,positive predictive value ,negative predictive value ,Youden index and diagnostic accuracy of PCT ,CRP ,PCT /CRP series test and parallel test were calculated for bacterial infectious pneumonia diagnosis .Results The levels of PCT and CRP of children in bacterial infectious pneumonia group were significant higher than those of children in non‐bacterial infectious pneumonia group and non‐infectious disease group(P < 0 .05) .The sensitivity ,specificity ,positive predictive value ,negative predictive value , Youden index and diagnostic accuracy of PCT ( ≥ 0 .5 ng/mL as the best intercept point) for bacterial infectious pneumonia diagno‐sis were 0 .743 ,0 .719 ,0 .806 ,0 .639 ,0 .461 and 0 .733 .And the same values for PCT /CRP series test were 0 .604 ,0 .875 ,0 .884 , 0 .583 ,0 .479 and 0 .709 respectively .All the values were higher than those of children in non‐bacterial infectious pneumonia group and non‐infectious disease group except sensitivity .Conclusion The combination of PCT and PCT /CRP series test is ideal method for early diagnosis of bacterial infectious pneumonia with high sensitivity and specificity .

Objective To estimate the biological effects of leptin on human HaCaT keratinocytes and explore their molecular mechanisms.Methods Cell counting kit-8 (CCK-8) was used to evaluate the proliferation of cultured HaCaT cells treated with different concentrations of leptin for 24 and 48 hours.Some HaCaT cells were classified into four groups to remain untreated,be treated with leptin (100 μg/L) and piceatannol (a specific inhibitor of STAT3 phosphorylation) alone or in combination for 24 hours,respectively,followed by the evaluation of cell proliferation using CCK-8 kit.Flow cytometry was performed to assess cell cycle of HaCaT cells treated with leptin of 100 μg/L,Western blot to determine the phosphorylation level of Erk1/2 and STAT3 in HaCaT cells treated with leptin of 100 μg/L for different durations.Statistical analysis was done by Student's t-test for unpaired data using GraphPad Prism 5 software.Results The proliferation of HaCaT cells was accelerated to different degrees after treatment with leptin of 50 and 100 μg/L for 24 and 48 hours,and the accelerating effect was in a dose-dependent manner within 24 hours (r =0.9989,P ＜ 0.05).Piceatannol apparently inhibited the promotive effect of leptin on the proliferation of HaCaT cells.There was an obvious elevation in the percentage of cells at S phase ((57.70 ± 5.88)％ vs.(42.50 ± 7.55)％,P ＞ 0.05),but a significant decrease in that at G0/G1 phase ((39.70 ± 1.57)％ vs.(45.20 ± 1.44)％,P ＜ 0.05),with a significant increase in proliferation index (0.603 ±0.0157 vs.0.564 ± 0.0144,P ＜ 0.05) in HaCaT cells treated with leptin of 100 μg/L for 24 hours compared with the untreated controls.Western blot showed that leptin of 100 μg/L markedly enhanced the phosphorylation level of STAT3 in HaCaT cells.Conclusion Leptin may upregulate the proliferation of HaCaT cells through activation of STAT3 pathway.