Objective To investigate the clinical efficacy of fractionated ultrapulse carbon dioxide laser combined with microneedle for depressed acne scars.Methods Totally 86 patients with depressed acne scar were randomly divided into 2 groups:the treatment group received fractionated ultrapulse carbon dioxide laser combined with microneedle technology and the control group only received fractionated ultrapulse carbon dioxide laser.The treatment interval was 1 to 3 months,and the whole course was 2-8 times.The total effective rate and the adverse reaction were analyzed in the facial acne scar.Results The surface of depressed acne scars was significantly improved.ECCA score of all patients declined obviously after treatment,the score of the treatment group declined from 61± 11 to 45± 15,and the control group declined from 57± 14 to 40± 16.There were significantly statistical differences in the results of each group (P＜0.05).More treatment times resulted in better surface improvement.The total effective rate were 81.8％ (18/22) and 100％ (24/24) in the treatment group,and 80.0％ (16/20) and 100％ (20/22) in the control group.The course was 6.8± 1.6 months in the treatment group and 20.8±4.6 months in the control group (P＜0.05) to get a similar satisfied result.There were no obvious adverse reactions except two cases of erythema and pigmentation in the treatment group and 5 cases in the control group.The former was with less adverse reactions.Conclusions Ultrapulse carbon dioxide laser combined with microneedle technique is effective for facial acne scars,with less complications and shorten treatment course.

Objective To investigate the clinical efficacy of Q-switch 1064 nm wavelength laser combining with microneedle technology for the treatment of melasma.Methods 70 melasma patients were selected in the Out-patient Department of Wuhan General Hospital of Guangzhou Military Command from July 2012 to Sept.2013.The patients were randomly divided into 3 groups:the combining therapy group (patients in this group received Q-switch 1064 nm wavelength laser combining with microneedles technology,the treatment interval was 2 or 3 weeks,and the whole course was 2-6 months);the microneedle group and the Q-switch 1064 nm wavelength laser group.The effect and the adverse reaction were observed and recorded during the treatment,and the reduced area of the pigment patch was also measured to judge the total efficacy after the course.Results The total effective rate was 78.6％ (22/28) in the combining therapy group and 40.0％ (8/20) in the microneedle group,45.5％ (10/22) in the Q-switch 1064 nm wavelength laser group.There were statistically significant differences between the combining therapy group and the other groups (P1 ＜0.05,P2 ＜0.05),without obvious adverse reaction.Conclusions It is significantly better to use American Medlite C6 Q-switch 1064 nm wavelength laser combining with microneedles technology than that of the single use of Q-switch 1064 nm wavelength laser or microneedles in the treatment for the patients with melasma.The method is also simple to handle and suitable for clinical application.

Objective To search for the method used in enriching caffeates in Cirsium setosum by macroporous adsorption resin, 23 types of macroporous adsorption resin were optimized, which could provide the industrial production of caffeates with the theory basis. Methods The adsorption and elution ratios, product purity and yield, as total indices, were comprehensively evaluatued by 23 types of macroporous adsorption resin in enriching the extracts in C. setosum. Results The product purity and yield of caffeates by the HPD-100 type macroporous adsorption resin were the highest and up to 52.2% and 87.6%. Conclusion The HPD-100 type macroporous adsorption resin shows better comprehensive adsorption property. It is available for the enrichment of caffeates in C. setosum.

OBJECTIVETo explore the clinical efficacy and action mechanism of acupuncture for premature ovarian failure (POF).

METHODSAccording to prospective cohort study design, 30 cases were included. Based on theory of regulating Chong and Conception Vessels as well as soothing the liver and calming the nerves, acupuncture at Guanyuan (CV 4), Dahe (KI 12), Taixi (KI 3), Taichong (LR 3), Baihui (GV 20) was applied, three times per week and 3 months were considered as one session. Totally two sessions were performed. The menstruation condition, estradiol (E2), follicle-stimulating hormone (FSH) and scores of the clinical perimenopausal symptoms were taken as treatment outcomes.

RESULTSThe total effective rate was 86.7% (26/30) and the cured rate was 16.7% (5/30). The recovery rate of menstruation was 16.7% (5/30) and the regain rate of menstruation was 56.7% (17/30). After the treatment, the level of E2 was significantly increased from (45.41 +/- 18. 09) pmol/L to (59.07 +/- 24.21) pmol/L (P < 0.05), and the score of perimenopausal symptoms was obviously reduced from 14.28 +/- 8. 30 to 4.04 +/- 3.28 (P < 0.01). However, no statistical differences could be seen in FSH before and after treatment (P > 0.05).

CONCLUSIONAcupuncture has certain effect on improving menstruation and perimenopausal symptoms in POF patients, which is likely to he related with increasing the level of E2.

Acupuncture Therapy ; Adolescent ; Estradiol ; metabolism ; Female ; Humans ; Luteinizing Hormone ; metabolism ; Primary Ovarian Insufficiency ; metabolism ; therapy ; Prospective Studies ; Young Adult

The quality of guidelines of clinical practice in acupuncture and moxibustion was evaluated with AGREE II tool. After retrieval of officially-issued guidelines of clinical practice in acupuncture and moxibustion from 2004 to 2013, totally 5 types of evidence-based guidelines of clinical practice in acupuncture and moxibustion were acquired, including depression, herpes zoster, pseudobulbar palsy after stroke, migraine and Bell's palsy. With AGREE II tool, independent score in six areas, including guideline's scope and purpose, participants and personnel, preciseness, clarity and readability, applicability and editorial independence were analyzed to perform a comprehensive evaluation in the end. The assessment results indicated that the score of editorial independence in evidence-based guidelines of clinical practice in acupuncture and moxibustion was the highest, averagely 97.9%, which was followed by clarity and readability (83.3%), participants and personnel (78.1%), preciseness (75.6%), the scope and purpose (68.1%) and the applicability (62.5%). The total evaluation score was all 6 points in the five guidelines with recommended as comprehensive evaluation advice. This study results indicate that although the guideline of clinical practice in acupuncture and moxibustion is low in number, the total quality is considerable.
Acupuncture Therapy ; standards ; Evidence-Based Medicine ; Humans ; Moxibustion ; standards ; Practice Guidelines as Topic ; Quality Assurance, Health Care

Objective: To investigate plasma interleukin-35 (IL-35) expression in sepsis patients, and to assess the regulatory activity of IL-35 on CD4⁺ and CD8⁺ T cells in sepsis patients. Methods: A prospective study was conducted. Forty-one sepsis patients and nineteen healthy controls were enrolled in this study. According to the survival outcome on 28 day after admission, sepsis patients were further divided into the survival group (n=30) and death group (n=11). Peripheral blood samples were collected within one hour of visit. Plasma IL-35 level was measured by enzyme linked immunosorbent assay (ELISA), while CD3+, CD4⁺, and CD8⁺ T cell counts were measured by flow cytometry. The correlation between IL-35 expression/T cell counts and sequential organ failure assessment (SOFA) score was also assessed. Peripheral CD4⁺ and CD8⁺ T cells from sepsis patients were purified and stimulated with recombinant human IL-35 for 48 h. mRNA relative levels of T-bet, GATA-3, FoxP3, and RORγt in CD4⁺ T cells and perforin, granzyme B, and FasL in CD8⁺ T cells were measured by real-time PCR. Interferon-γ (IFN-γ), IL-4, IL-10, IL-17, and tumor necrosis factor-α (TNF-α) levels in the cultured supernatants were measured by ELISA. Transcription factor level and cytokine expression was compared prior to and post IL-35 stimulation. Results: Plasma IL-35 was significantly elevated in sepsis patients when compared with controls [(76.76±10.33) pg/mL vs (27.53±8.31) pg/mL, P<0.01], however, it did not correlate with SOFA score (r=0.172, P=0.281). Plasma IL-35 was also notably increased in the death group when compared with the survival group [(83.18±6.48) pg/mL vs (74.40±10.55) pg/mL, P=0.014]. Plasma IL-35 level in the death and survival groups were analyzed by receiver operating characteristic curve with an area under curve of 0.770 (P=0.009). There were no remarkable differences of CD3⁺, CD4⁺, and CD8⁺ T cell counts between sepsis patients and controls, or between the death and survival groups (all P>0.05). Moreover, T cell counts were negatively correlated with SOFA score (P<0.01). IL-35 stimulation resulted in the reduction of mRNA relative level of Th1 transcription factor T-bet and Th17 transcription factor RORγt in CD4⁺ T cells from sepsis patients (P<0.01), while elevation in regulatory T cell transcription factor FoxP3 mRNA (P<0.01). However, IL-35 stimulation did not affect Th2 transcription factor GATA-3 mRNA in CD4⁺ T cells (P=0.745). IFN-γ and IL-17 production by CD4⁺ T cells was significantly decreased in response to IL-35 stimulation (P<0.05), while IL-10 was increased (P<0.01) and IL-4 was comparable (P=0.536) between CD4⁺ T cells with and without IL-35 stimulation. IL-35 stimulation led to the down-regulation of mRNA relative level of perforin and granzyme B (P<0.01). However, FasL mRNA was comparable between CD8⁺ T cells with and without IL-35 stimulation (P=0.795). IFN-γ and TNF-α secretion was also reduced in response to IL-35 stimulation (P<0.05). Conclusion Elevated plasma IL-35 level is an important indicator for poor prognosis of sepsis patients. IL-35 inhibits the activity of CD4⁺ and CD8⁺ T cells in sepsis patients, and might take part in the pathogenesis of sepsis.

Objective During pregnancy , exosomes can be released from the placenta into maternal circulation and play im-portant roles in normal pregnancy or placenta-related diseases .We aimed to establish a simple and efficient method for isolating and i-dentifying placental exosomes from maternal serum and lay a foundation for the studies of pregnancy -related diseases . Methods Using sucrose gradient centrifugation with 8% PEG6000 precipitation twice , we isolated and purified placenta-derived exosomes from normal maternal serum and detected their molecular markers CD 63 , CD81 and PLAP by Western blot , followed by silver staining anal-ysis of the protein profile of the exosome pellet .We identified the morphology of the placenta-derived exosomes by transmission electron microscopy ( TEM) and measured the size and distribution of the particles by dynamic light scattering ( DLS) . Results Silver stai-ning of the protein profiles of the exosomes after sucrose gradient centrifugation clearly revealed the bands of the protein molecules . Western blot showed the expressions of CD 63, CD81, and PLAP in the 21-34%density layer, which demonstrated the presence of serum placental exosomes mainly in the 1.09-1.16 g/mL density layer.TEM exhibited that the placenta-derived exosomes were round or oval cup-shaped, specifically expressing PLAP, and the particles were uniform in size, with a mean diameter of (41.79 ±11.94) nm. Conclusion A simple, fast, and efficient method was successfully established for isolating placenta-derived exosomes from ma-ternal serum, which provides a basis for studying the roles of placental exosomes in normal pregnancy and placenta -related diseases.

Objective:To analyze the polymorphism in human cDNA sequence of prothymosin-?(ProT?)by sequencing analysis.Methods:The cDNA of human ProT? was amplified from cells of peripheral blood and cord blood by RT-PCR.The product of RT-PCR was purified and linked with vector pMD18-T.After cloning and sequencing,the sequence of ProT? cDNA was compared with the standard sequence to analyze the polymorphism in the ProT? cDNA sequence.Results:The cloned ProT? cDNA sequence was different from that of the standard.We found 2 kinds of variations:(1)The nucleotide in 107 position was varied and the nucleotides in 110-121 and 191-205 positions were deleted;(2)The nucleotide in 306 position was deleted,mainly in the 60-80 years old group.Conclusion:We have identified 2 kinds of variations in human ProT? cDNA,but the first 28 amino acid in the N-terminal of cDNA of human ProT? are not involved therefore the variations do not affect the function of human ProT?.

M2e gene of three copies for H5N1 subtype AIV was synthesized and fused with human mycobacterium tuberculosis hsp70 gene.The fused gene was cloned into the prokaryotic expression vector to get pET-3M2e and pET-3M2e-hsp70.Recombinant protein r3M2e and r3M2e-hsp70 were successfully expressed induced with IPTG and purified with Ni2+-NTA collumn.Following that,the immunity of the recombinant protein was analysized with Western blot.20-day-old AIV non-immunized chickens were vaccined with r3M2e and r3M2e hsp70,at the same time,Trx and KLH-M2e inoculated chickens were served as vector and positive controls.Two weeks after the primary vaccination,every group was boosted with the same vaccine as in the primary vaccination.The humoral immunity of the vaccined chickens was evaluated with antibody detection against M2e,cytopathic suppression test,and indirect fluorescence assay.The cellular immunity was estimated according to lymphocyte subtype analysis with flow cytometry and M2e specific cytokine detection.Four weeks after the boost vaccine,all groups were challenged with 100EID50 AIV of H9N2 subtype,and the virus from swabs was detected with Real-time PCR.Results indicated that r3M2e hsp70 vaccined chicken developed the better humoral and cellular immune response,also,made a better performance compared with r3M2e vaccined group in virus challenge.

Objective To evaluate the effect of hydrogen peroxide (H2O2) on autophagy in melanocytes,and to explore its possible regulatory mechanisms.Methods Normal human melanocytes at exponential growth phase were divided into several groups:blank control group receiving no treatment,positive control group treated with 100 nmol/L sirolimus solution,and experiment groups treated with H2O2 solution at different volume fractions of 10-7-10-3 respectively.After 4-hour treatment,cell counting kit-8 (CCK-8) assay and flow cytometry were performed to evaluate the cellular proliferative activity and detect apoptosis of melanocytes respectively.Acridine orange staining was performed to detect autophagosome formation,transmission electron microscopy to observe ultrastructural changes of autophagosomes,and Western blot analysis to measure the expression of autophagy-specific protein Beclin 1 and microtubuleassociated protein 1 light chain 3B (LC3B).A total of 84 autophagy-related genes were analyzed by RT2 Profiler PCR Array,so as to screen differentially expressed autophagy-related genes.Results After the treatment with H2O2 at different volume fractions of 10-3,5 × 10-4,10-4,5 × 10-5,10-5,5 × 10-6 and 10-6,experiment groups showed significantly decreased cellular proliferative activity,but significantly increased apoptosis rate compared with the blank control group (F =286.95,301.23,respectively,both P ＜ 0.05).With the increase in volume fractions of H2O2,the cellular proliferative activity was significantly gradually decreased (P ＜ 0.05),while the apoptosis rate showed an opposite trend (P ＜ 0.05),except that the 5 ×10-6 H2O2 group showed no significant differences in the apoptosis rate compared with the 10-5 H2O2 group and 10-6 H2O2 group.Acridine orange staining and electron microscopy showed autophagosome formation in the 10-5 H2O2 group,10-6 H2O2 group and positive control group.Western blot analysis revealed that Beclin1 expression and LC3B-Ⅱ/LC3B-Ⅰ ratio were significantly higher in the 10-5 H2O2 group,10-6 H2O2 group and positive control group than in the blank control group (all P ＜ 0.05).RT2 Profiler PCR Array showed significant up-regulation of ATG12,ATG3,ULK1,PIK3CG,PTEN and PIK3C3 genes and significant downregulation of EIF2AK3 gene in the 10-5 H2O2 group,10-6 H2O2 group and positive control group compared with the blank control group.In the 10-5 H2O2 group and positive control group,the mTOR gene was significantly up-regulated,and the ULK2 gene was significantly down-regulated.The 10-6 H2O2 group showed no obvious changes in the expression of mTOR gene,but significant up-regulation of AMPK and JNK1 genes.Conclusion H2O2 at volume fractions of 10-5 and 10-6 can induce autophagy in melanocytes,likely by influencing the expression of some related signaling molecules.