Objective To determine the expression of miRNA211 (miR-211) in the development of malignant melanoma,and to investigate the correlation between miR-211 and its target molecule,matrix metalloproteinase 16 (MMP-16).Methods Cultured A375 melanoma cells were divided into 3 groups:miR-211 overexpression group and mock-vehicle group transfected with miR-211 mimics and empty vehicle respectively,and negative control group receiving no treatment.TaqMan fluorescence-based quantitative PCR was performed to determine the expression of miR-211 in HER1 primary melanocytes,A375,C32 and G361malignant melanoma cell lines,as well as in nevus tissues (n =18) and melanoma tissues (n =41),and to evaluate changes of MMP-16 mRNA expression in A375 cells before and after the overexpression of miR-211.Sulforhodamine B (SRB) assay and flow cytometry were conducted to evaluate cellular proliferative activity and determine cell cycle distribution respectively,and methylcellulose assay and Transwell assay to evaluate colony formation and cell migration abilities respectively.The size of selected colonies was used to represent colony formation ability,while the ratio of the number of migrating cells to that of non-migrating cells to represent cell migration ability.Results There were significant differences in the expression level of miR-211 among the G361,C32 and A375 cells (0.09 ± 0.02 vs.0.000 52 ± 0.000 20 vs.0.000 03 ± 0.000 01,F =10 410,P ＜ 0.01).The expression of miR-21 1 was significantly decreased in melanoma tissues compared with nevus tissues (0.17 ± 0.03 vs.0.87 ± 0.08,t =9.118,P ＜ 0.01).No significant differences were observed in cellular proliferative activity or cell cycle distribution among the miR-211 overexpression group,mock-vehicle group and negative control group.Compared with the mock-vehicle group,the miR-211 overexpression group showed significantly suppressed colony formation (0.49 ± 0.05 vs.0.85 ± 0.09,t =2.19,P ＜ 0.05) and cell migration (0.49 ± 0.06 vs.0.82 ± 0.09,t =3.15,P ＜ 0.05) abilities,while no significant difference was observed between the mock-vehicle group and negative control group.Additionally,the mRNA expression of MMP-16 significantly decreased in the miR-211 overexpression group compared with the mock-vehicle group after transfeetion (24 hours:0.33 ± 0.02 vs.0.91 ± 0.03,t =11.30,P ＜ 0.01;48 hours:0.52 ± 0.01 vs.0.96 ± 0.02,t =5.02,P ＜ 0.05;72 hours:0.71 ± 0.01 vs.0.97 ± 0.03,t =3.85,P ＜ 0.05),with no significant difference between the mock-vehicle group and negative control group at the above time points.Conclusions miR-211 was lowly expressed in both malignant melanoma cells and tissues,and it could inhibit both anchorage-independent growth and migration of melanoma cells.After up-regulation of miR-211 expression,the mRNA expression of MMP-16 decreased in A375 cells,suggesting that MMP-16 may be a downstream target of miR-211,and can influence melanoma metastasis.

The application of placebo acupuncture in randomized controlled trials in the past 10 years (2004-2014) in foreign countries was systematically reviewed to summarize the design of placebo acupuncture; according to category of diseases, advantages and disadvantages were analyzed to explore an ideal placebo acupuncture set. By retrieval in PubMed, EMBASE and OVID databases, the clinical study literature which met the inclusion criteria was searched, and the category of diseases, design of placebo acupuncture and clinical efficacy were analyzed. Totally 29 articles were included. The pain was the leading disease in category of diseases; the most commonly used application of placebo acupuncture design was telescopiform sliding blunt needle, accounting for 45. 16%; the treatment locations were non-acupoints mostly; in the needling depth, approximately 60. 00% selected non-penetrating needling and 26. 67% selected superficial needling; 28 articles indicated that acupuncture and placebo acupuncture had clinical effects, accounting for 96. 55%; 37. 93% of articles indicated acupuncture was superior to placebo acupuncture and 37. 93% of articles indicated acupuncture was not superior to placebo acupuncture. It is concluded that the design of placebo acupuncture should consider multiple factors, including main symptoms, operability, security, blinding and specific effects, and the objective indices should be selected for outcome evaluation.
Acupuncture Therapy ; instrumentation ; methods ; standards ; Humans ; Internationality ; Needles ; Placebo Effect ; Randomized Controlled Trials as Topic ; standards

Objective To observe the therapeutic effect of electronic moxibustion apparatus and stone needle for primary dysmenorrhea disease in 35 cases.Methods 70 patients were randomly divided into the treatment group of 35 cases by random number table ,using electronic moxibustion apparatus and stone needle for treatment;and con-trol group of 35 cases,only using electronic moxibustion apparatus for treatment from 5 days before menstruation ,the treatment was continuous for 5 days,a course including 3 menstruation cycles .The clinical therapeutic effect after one treatment course was observed .Results The curative effects of the two groups were respectively 91.42% and 88.57%(χ2 =0.16,P>0.05);recovery rates of the two groups were respectively 77.14%and 42.85%(χ2 =8.57, P<0.05).The scores of the VAS,CMSS of the treatment group were much lower than those of the control group after treatment (t=2.532,2.137,2.256,P<0.05).Conclusion Electronic moxibustion apparatus and stone needle therapy is an effective method to treat primary dysmenorrhea ,which is a safe and convenient method .

Objective To explore the effect of human leukocyte antigen B* genotype and age on serum homocysteine (Hcy) levels in children with seizures or epilepsy. Methods Fifteen children with seizures or epilepsy in whom HLA-B*15:02 genotype was detected during October 2015 to June 2016 were included. The plasma Hcy concentration in children with different genotypes was compared. The association of Hcy concentration and age was performed by linear-regression analysis. Results The mean concentration of Hcy was 8.38±4.23 μmol/L in children not carrying HLA-B*15:02 gene, which was obviously higher than that in children carrying HLA-B*15:02 gene 13.03±0.97 μmol/L (P<0.05). The Hcy concentration increased with the age (r2 =0.29, P<0.05). Conclusions Elder children with seizures or epilepsy carrying HLA-B*15:02 gene tend to have higher Hcy concentration and increased potential risk of disease. HLA-B*15:02 gene type and age can predict the changes of Hcy concentration in children with convulsions.

Background: Phototherapy is an important method to treatvitiligo. However, it is unclear how phototherapy affectsmelanocyte precursors and skin neural crest stem cells. Objective: To investigate the underlying mechanisms of narrow-band ultraviolet B (NB-UVB) induced melanocyte lineagedifferentiated from human scalp-derived neural creststem cells (HS-NCSCs). Methods: HS-NCSCs were expandedfrom scalp hair follicles. The c-Kit−/CD57− HS-NCSCs wereisolated by cell sorting. Different doses of NB-UVB wereused to irradiate these HS-NCSCs. Cell ultrastructure was examinedby transmission electron microscope. Melanocytemarker expression was analyzed by Quantitative RT-PCRand Western blot. Cell proliferation and migration were alsoevaluated. Results: The c-Kit−/CD57− HS-NCSCs expressedembryonic NCSC biomarkers. NB-UVB at a dose of 100 mJof NB-UVB had little effect on the cell proliferation of differentiatedmelanocytes from c-Kit−/CD57− HS-NCSCs, while700 mJ inhibited cell proliferation significantly. The dendriticprocesses of differentiated melanocytes increased afterradiation. The tyrosinase and Melanocortin 1 receptor (Mc1R)expression of differentiated melanocytes increased after NB-UVB exposure. The effect of NB-UVB on tyrosinase expressionwas modulated by signaling inhibitors H89 andPD98059 as well as Mc1R level in the cells. The migrationability of differentiated melanocytes was enhanced under100 mJ exposure. Conclusion These data demonstrate thatNB-UVB facilitates melanocytic differentiation of the HSNCSCsand enhances migration of these cells. Mc1R andcAMP pathway play a critical role in NB-UVB induced melanocyticdifferentiation.

Objective To investigate the efficacy of lateral decubitus intramedullary nailing for treatment of subtrochanteric fractures of the femur.Methods From January 2012 to December 2015,23 patients with simple femoral subtrochanteric fracture were treated at Department of Orthopedic Trauma,Changhai Hospital.They were 15 males and 8 females,aged from 19 to 77 years (average,48.3 years).According to the Seinsheimer classification,there were 6 cases of type ⅡB,8 cases of type ⅡC,6 cases of type Ⅲ A,and 3 cases of type ⅢB.Their injuries were caused by traffic accident in 10 cases,falling from a height in 5 cases,and sprain in 8 cases.All patients were treated by closed reduction and anterograde intrarnedullary nailing at lateral decubitus.Their operative time,bleeding volume,fluoroscopic frequency,fracture healing time,functional recovery and complications were recorded and analyzed.Results Their operative time ranged from 55 to 80 min,averaging 65.7 min;their bleeding volumes ranged from 240 to 420 mL,averaging 304.3 mL;their fluoroscopic frequency ranged from 30 to 60 times,averaging 42.7 times.This cohort was followed up for 12 to 28 months (average,17.9 months).Their fracture healing time ranged from 4 to 10 months,averaging 5.5 months.Nonunion occurred in one patient but was cured by secondary operation.The HSS evaluation at the final follow-ups showed 17 excellent cases and 6 good ones,yielding an excellent to good rate of 100％.All the wounds healed by the first intention.No infection,deep vein thrombosis or implant failure was observed.Conclusion As lateral decubitus intramedullary nailing can achieve satisfactory clinical efficacy for subtrochanteric fractures of the femur,the body position of lateral decubitus may be a good alternative.

Objective : To investigate the effects of hypothyroidism and drug intervention on the cognitive function of rat offspring through the establishment of a rat model of experimental hypothyroidism in pregnancy. Methods : A total of 20 SD female rats were randomly divided into control group ( CON group) and hypothyroid group ( PTU group) . The hypothyroid model was established by propylthiouracil ( PTU) , the thyroid hormone levels of female mice were detected by electrochemiluminescence immunoassay , and the differences between the two groups were compared. After successful modeling , the male mice were mated in cages , and the hypothyroid group was randomly divided into no intervention group ( group I) , first trimester intervention group ( group II) and second and third trimester intervention group ( group III) . The Morris water maze ( MWM) experiment was used to test the learning and memory ability of the rats. The morphological structure of hippocampal neurons , the expression of nucleoprotein (NeuN) and synapse⁃associated protein ( SYN) in mature neurons were observed by hematoxylin⁃eosin staining (HE) , Nissl staining and immunohistochemistry. Results : ① Compared with the CON group , the female mice in the PTU group had a significant increase in TSH and a significant decrease in FT4 (P < 0. 05) . ② In the positioning navigation test , the evasion latency of the pups in each group was gradually shortened. On the 5th day , the incubation period of group I was significantly longer than that of groups CON , II , III (P < 0. 05) . There was little change between groups II and III and CON groups (P > 0. 05) . ③ The residence time of group II was significantly different from that in group I during the space exploration stage (P < 0. 05) . There was a significant difference between the number of pups crossing the platform in group I and groups CON , II and III (P < 0. 05) . ④ There was no significant difference between HE staining and Nissl staining in hippocampal tissues of rats in each group. However, compared with the CON group , the average absorbance of NeuN and SYN proteins in the hippocampus of mice in groups I , II and III was significantly reduced (P < 0. 05) . Conclusion Hypothyroidism will have adverse effects on the cognitive function and hippocampal neuron development of pregnant rats , and the effects of intervention on the cognitive function of offspring at different stages of pregnancy are different , the earlier the intervention , the smaller the damage to cognitive function.

BackgroundSchizophrenia is a common severe mental disorder with complex pathogenesis. There are few studies on the correlation between kynurenine metabolites in peripheral serum and urine in schizophrenia. ObjectiveTo investigate the concentration of tryptophan-kynurenine metabolites and interleukin-6 （IL-6） in serum and urine in patients with schizophrenia， and their correlation with clinical symptoms， so as to explore potential biological characteristics related to schizophrenia. MethodsA total of 38 patients with schizophrenia who met the criteria of the Diagnostic and Statistical Manual of Mental Disorders， fifth edition （DSM-5）， and were hospitalized or attended outpatient clinic at Hangzhou Seventh People's Hospital from December 2021 to December 2022 were included in the study. Additionally， 26 healthy individuals were concurrently recruited from the community of Hangzhou to serve as a control group. All participants were requested to complete the Positive and Negative Symptom Scale （PANSS）. The levels of tryptophan （TRP）， kynurenine （KYN）， kynurenic acid （KYNA）， quinolinic acid （QUIN）， picolinic acid （PIC）， xanthurenate and 5-hydroxytryptamine （5-HT） in both serum and urine were measured using ultra-high-performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry. Serum and urine IL-6 were measured using enzyme-linked immunosorbent assay. Pearson correlation analysis was conducted to examine the correlation between serum and urinary KYN metabolites， as well as the correlation between metabolite levels and clinical symptoms in the patient group. ResultsPatients with schizophrenia had significantly higher level of IL-6 in serum （U=798.500， P<0.01） and lower level of PIC in urine （U=253.000， P=0.013） compared with the control group. Additionally， level of serum KYN was positively correlated with QUIN/KYNA ratio and QUIN/PIC ratio （r=0.562， 0.438， P<0.05） in patients with schizophrenia. 5-HT/KYN ratio in serum was positively correlated with PANSS total score and negative symptom subscale score （r=0.458， 0.455， P<0.01） in patients with schizophrenia. ConclusionSerum TRP-KYN pathway metabolite levels in patients with schizophrenia were associated with neurotoxic metabolite ratios in urine and the severity of negative symptoms. ［Funded by Zhejiang Medical and Health Science and Technology Program Exploratory （number， 2022KY990）］

OBJECTIVETo compare the effects between acupuncture with regulating mind and spleen and wes-tern medication for diarrhea irritable bowel syndrome (IBS-D).

METHODSEighty-one patients were randomly at the ratio of 2 to 1 assigned into an acupuncture group (54 cases) and a western medication group (27 cases). Acupuncture with regulating mind and spleen was applied in the acupuncture group for 6 weeks at Baihui (GV 20), Yintang (GV 29), Tianshu (ST 25), Zusanli (ST 36), Shangjuxu (ST 37), Sanyinjiao (SP 6), and Taichong (LR 3), once every other day, 3 times a week. Pinaverium bromide tablet was used orally in the western medication group for 6 weeks, 50 mg a time, 3 times a day. IBS symptom severity score (IBS-SSS) were observed before and after 1-week, 6-week treatment. Pittsburgh sleep quality index (PSQI) was applied before and after 6-week treatment. Also, clinical efficacy was evaluated in the two groups.

RESULTSThree patients dropped out in the acupuncture group, and 1 in the western medication group. Except abdominal distension score after 1-week treatment in the western medication group, single scores and total scores of IBS-SSS apparently reduced in the two groups after 1-week and 6-week treatment (<0.01,<0.05). After 1-week treatment, the abdominal pain score of the acupuncture group was obviously lower than that of the western medication group (<0.05). After 6-week treatment, with abdominal pain relief advantage, other results including the number of pain days, defecation satisfaction,life disturbance degree and total score of the acupuncture group were obviously lower than those of the wes-tern medication group (<0.01,<0.05). After 6-week treatment, the PSQI score and its change before and after treatment in the acupuncture group were superior to those in the western medication group (both<0.05). The relief rate and relief plus obvious effective rate in the acupuncture group were higher than those in the western medication group[51.0% (26/51) vs 19.2% (5/26),<0.01; 64.7% (33/51) vs 34.6% (9/26),<0.05].

CONCLUSIONSAcupuncture with regulating mind and spleen for diarrhea irritable bowel syndrome can more effectively relieve abdominal pain than pinaverium bromide tablet at the early stage. Its total effect and single effects are better at the later stage on abdominal pain, seizure frequency, defecation satisfaction, life disturbance, and sleep quality.