The Therapeutic Effects of Exosomes Derived from Human Umbilical Cord Mesenchymal Stem Cells on Scleroderma

BACKGROUND@#Scleroderma is a multisystem disease in which tissue fibrosis is caused by inflammation and vascular damage. The mortality of scleroderma has remained high due to a lack of effective treatments. However, exosomes derived from human umbilical cord mesenchymal stem cells (HUMSCs)-Ex have been regarded as potential treatments for various autoimmune diseases, and may also act as candidates for treating scleroderma. @*METHODS@#Mice with scleroderma received a single 50 lg HUMSCs-Ex. HUMSCs-Ex was characterized using transmission electron microscopy, nanoparticle tracking analysis and nanoflow cytometry. The therapeutic efficacy was assessed using histopathology, immunohistochemistry, immunofluorescence, quantitative real-time polymerase chain reaction, enzyme-linked immunosorbent assay and western blot. @*RESULTS@#HUMSCs-Ex ameliorated the deposition of extracellular matrix and suppressed the epithelial-mesenchymal transition process, and the effects lasted at least three weeks. In addition, HUMSCs-Ex promoted M1 macrophage polarization and inhibited M2 macrophage polarization, leading to the restoration of the balance of M1/M2 macrophages. @*CONCLUSION@#We investigated the potential antifibrotic and anti-inflammatory effects of HUMSCs-Ex in a bleomycininduced mouse model of scleroderma. So HUMSCs-Ex could be considered as a candidate therapy for scleroderma.

Catwalk gait analysis for evaluating any effect of transcranial pulsed current stimulation on motor functioning after a stroke / 中华物理医学与康复杂志

Objective:To evaluate any effect of transcranial pulsed current stimulation (tPCS) on the motor functioning of rats modelling stroke using the Catwalk gait analysis system.Methods:A stroke model was induced in 24 rats using middle cerebral artery embolization. They were then randomly divided into a sham operation group, a model group and a tPCS group, each of 8. Neurological deficit scores were assigned 1 day after the modeling. Beginning two days after the modeling the tPCS group was given 20 minutes of tPCS daily with an intensity of 0.2mA at 10Hz for 7 days. Gait data were collected using the Catwalk gait system 1 day before, as well as 1 and 9 days after the modeling.Results:Nine days after the modeling the average Bederson neuroethology score of the tPCS group was significantly lower than one day after the modelling and significantly lower than the model group′s average. One day after the modelling significant differences were observed in the model and tPCS groups in the average contact area of the affected limb′s paw prints, limb swing speed, stride length, limb speed, swing time, average running speed and standing time compared with before the operation. After nine days the average standing time on the affected fore and hind limbs, as well as the paw contact areas were significantly better in the tPCS group than in the model group.Conclusion:tPCS can promote improvements in gait after ischemia and reperfusion, at least in rats.

Genetic analysis of a pedigree affected with tuberous sclerosis complex caused by a novel mutation of TSC1 / 中华医学遗传学杂志

Objective@#To analyze the clinical phenotype of a Chinese pedigree affected with Tuberous sclerosis complex(TSC) and explore pathogenic mutations of TSC1 and TSC2 gene.@*Methods@#Unique clinical phenotypes, the results of imaging, examination of the proband and special family history, collectively, made the constellation of features of TSC.Genomic DNA was obtained from six affected and eight unaffected members of the family and potential mutations of the TSC1 and TSC2 genes were detected by PCR-amplification of the exons and exon-intron boundaries and direct sequencing.A total of 150 normal unrelated individuals were used as controls.@*Results@#Genetic analysis documented the presence of a heterozygous mutation, c. 1781_1782delTG (p.Val594GlyfsX11), in the exon 15 of TSC1 gene within all the patients of the family. This mutation was not observed in the eight unaffected family members or in the 150 unrelated control subjects from the same population , or the Human Gene Mutation Database(HGMD)and had completely co-segregated with the disease phenotype in the family.@*Conclusions@#The c. 1781_1782delTG mutation of TSC1 gene may be responsible for the tuberous sclerosis complex in this family. The data presented in the present study are of significance to clinicians, as well as genetic counselors, and may provide new clues for molecular diagnosis of this disease.

Genetic analysis of a pedigree affected with tuberous sclerosis complex caused by a novel mutation of TSC1 / 中华医学遗传学杂志

OBJECTIVE@#To analyze the clinical phenotype of a Chinese pedigree affected with Tuberous sclerosis complex (TSC) and explore pathogenic mutations of TSC1 and TSC2 gene.@*METHODS@#Unique clinical phenotypes，the results of imaging, examination of the proband and special family history, collectively, made the constellation of features of TSC. Genomic DNA was obtained from six affected and eight unaffected members of the family and potential mutations of the TSC1 and TSC2 genes were detected by PCR-amplification of the exons and exon-intron boundaries and direct sequencing. A total of 150 normal unrelated individuals were used as controls.@*RESULTS@#Genetic analysis documented the presence of a heterozygous mutation, c.1781_1782delTG (p.Val594GlyfsX11), in the exon 15 of TSC1 gene within all the patients of the family. This mutation was not observed in the eight unaffected family members or in the 150 unrelated control subjects from the same population , or the Human Gene Mutation Database (HGMD) and had completely co-segregated with the disease phenotype in the family.@*CONCLUSION@#The c.1781_1782delTG mutation of TSC1 gene may be responsible for the tuberous sclerosis complex in this family. The data presented in the present study are of significance to clinicians, as well as genetic counselors, and may provide new clues for molecular diagnosis of this disease．.

Epidemiology of fungal infections in China / 医学前沿

With the increasing number of immunocompromised hosts, the epidemiological characteristics of fungal infections have undergone enormous changes worldwide, including in China. In this paper, we reviewed the existing data on mycosis across China to summarize available epidemiological profiles. We found that the general incidence of superficial fungal infections in China has been stable, but the incidence of tinea capitis has decreased and the transmission route has changed. By contrast, the overall incidence of invasive fungal infections has continued to rise. The occurrence of candidemia caused by Candida species other than C. albicans and including some uncommon Candida species has increased recently in China. Infections caused by Aspergillus have also propagated in recent years, particularly with the emergence of azole-resistant Aspergillus fumigatus. An increasing trend of cryptococcosis has been noted in China, with Cryptococcus neoformans var. grubii ST 5 genotype isolates as the predominant pathogen. Retrospective studies have suggested that the epidemiological characteristics of Pneumocystis pneumonia in China may be similar to those in other developing countries. Endemic fungal infections, such as sporotrichosis in Northeastern China, must arouse research, diagnostic, and treatment vigilance. Currently, the epidemiological data on mycosis in China are variable and fragmentary. Thus, a nationwide epidemiological research on fungal infections in China is an important need for improving the country's health.

Transcutaneous vagus nerve stimulation for primary insomnia and affective disorder:a report of 35 cases / 中国针灸

<p><b>OBJECTIVE</b>To observe and evaluate the clinical efficacy of transcutaneous vagus nerve stimulation (taVNS) at auricular concha for primary insomnia (PI) and affective disorder.</p><p><b>METHODS</b>A total of 35 patients who met the diagnosis standard of PI in(5th edition) were included. The self-developed auricular vagus nerve stimulator (TENS-200A) was applied at auricular concha, 30 min per treatment, twice a day, 5 days a week for consecutive 4 weeks. The follow-up visit was conducted at the end of 6th week. The Pittsburg sleep quality index scale (PSQI), 17-items Hamilton depression scale (17HAMD) and Hamilton anxiety scale (HAMA) were applied for evaluation. The PSQI, HAMA and 17HAMD were observed before and after treatment; the safety was also observed.</p><p><b>RESULTS</b>Compared before treatment, the PSQI was significantly decreased to (13.20±3.61) at the end of 2nd week (<0.05); compared before treatment, the 17HAMD and HAMA were significantly decreased at the end of 4th week and 6th week (all<0.05). No adverse reaction was observed.</p><p><b>CONCLUSIONS</b>The taVNS could not only relieve PI symptoms, but also improve the depressive and anxiety symptoms, in addition, it may have positive long-term efficacy and safety.</p>

Effect of Acupuncture on Motor Function of Extrahepatic Biliary System of Guinea Pig / 世界科学技术-中医药现代化

This study was aimed to compare the effects of acupuncture stimulation on different acupoints, which included LR14-Qimen, ST25-Tianshu, GB34-Yanglingquanand CO11-pancreas and gallbladder (auricular vagus nerve intensively distributed areas) on gallbladder pressure and Oddi sphincter function of guinea pig, in order to discuss acupuncture effect on different acupoints in the regulation of motor function of extrahepatic biliary system. A total of 20 adult guinea pigs of both sexes were randomly divided into two groups, with 10 in each group. The detection was made on changes of Oddi sphincter electromyography and gallbladder pressure before and after acupuncture. A double-hook electrode was implanted into the Oddi sphincter for the detection of Oddi sphincter electromyography. And one small artificial balloon was put into the gallbladder to measure the intrabiliary pressure. After the guinea pig was anesthetized, changes of gallbladder pressure as well as the electromyography frequency and amplitude index of Oddi sphincter of the normal guinea pig were recorded. When the guinea pig was stabilized, acupuncture was given on LR14, ST25, GB34 and CO11 for 1 min. Observations were made on acupuncture effects of gallbladder pressure and Oddi sphincter function of guinea pig. The results showed that after acupuncture on LR14 and ST25, the electromyography frequency of Oddi sphincter was obviously increased compared with before acupuncture (frequency:P < 0.01, amplitude:P < 0.05); the gallbladder pressure was obviously decreased (P < 0.05). After acupuncture on CO11 and GB34, the electromyography frequency and amplitude index of Oddi sphincter were obviously decreased compared with before acupuncture (P < 0.05); the gallbladder pressure was obviously increased (P < 0.01). It was concluded that acupuncture on LR14 and ST25 can obviously inhibit the movement of gallbladder and promote the myoelectricity release of Oddi sphincter. Acupuncturing on CO11 and GB34 can apparently promote the shrinking movement of gallbladder and inhibit the myoelectricity release of Oddi sphincter.

Treatment of ankylosing spondylitis by fire-needle therapy plus tuina manipulations / 针灸推拿医学（英文版）

Objective:To observe the clinical effect of fire-needle therapy plus tuina manipulations for ankylosing spondylitis (AS). Methods:A total of 49 eligible cases were treated by fire-needle therapy plus tuina manipulations, once every other day. Ten days made one course. After the continuous treatment for six courses, the therapeutic effects were summarized and the changes in the clinical symptoms and signs were observed. Results:During treatment, 7 cases dropped off. In 42 patients who had completed treatment for six courses, the results showed clinical recovery in 16 cases, remarkable effect in 13 cases, effect in 7 cases and failure in 6 cases. The measurements of the patients were all improved in different degrees than those before treatment. Moreover, no side effect was noticed during treatment. Conclusion:The therapeutic effect is precise and free of adverse reaction in treatment of AS by fire-needle therapy plus tuina manipulations.

Current research situation and prospect of auricular acupuncture for epilepsy / 中国针灸

Epilepsy is a group of chronic diseases characterized by recurrent and transient brain dysfunction induced by abrupt and intermittent abnormal discharge of neurons in the brain, which is difficult to be cured. Thee auricular concha are is he zone of visera in TCM auricular points, but also the innervating area of the auricular brach of the vagus nerve. Auricular acupuncture, as a special treatment, has superior therapeutic effect for epilepsy. In recent years, remarkable progress has been made in the theoretical basis and clinic application of auricular acupuncture for epilepsy, and the ear-vagus percutaneous electrical stimulation apparatus (auricular acupuncture apparatus) is developed for clinical trial researches. From the basic study to clinical research of auricular acupuncture for epilepsy, the epilepsy pathogenesis as well as the intervention pathway and clinical efficacy of auricular: experimental support and rich clinical experience for auricular acupuncture apparatus.

The central mechanisms underlying the phenomenon of acupoint sensitization evoked by visceral nociceptive afferent / 中国针灸

The physiological mechanism underlying the acupoint sensitization was evaluated systemically by using the method of electric physiology at spinal cord, medulla, and thalamus levels; the dynamic change of acupoint from the relative "silence" to the relative "activation" function was explained through the study on the dynamic process of acupoint sensitization; the biological process of the therapeutic effect of acupoint stimulation was illuminated through the research of the central mechanism underlining the dose effect relationship between the sensitive acupoint and the related brain area, thus scientific evidence for the functional link between the acupoint and internal organs as well as the nature of the acupoint were provided.

CSP genotypes and antifungal susceptibility of Aspergillus fumigatus isolates from clinical settings in different regions of China / 中华皮肤科杂志

Objective To describe the CSP genotypic profile in clinical isolates of Aspergillus fumigates from different regions of China,and to investigate if there is a difference in antifungal susceptibility among A.fumigates of different CSP genotypes and from different regions.Methods Totally,112 A.fumigates strains clinically isolated from Fujian,Shanghai,Hebei and Beijing were included in this study,and identified according to macro-and micro-morphological characters,growth temperature and β-tubulin sequence.Classic A.fumigatus strains were typed according to CSP gene sequence.The minimal inhibitory concentrations (MICs) of voriconazole,itraconazole and amphotericin B to A.fumigates were determined in accordance with the National Committee for Clinical Laboratory Standards (NCCLS) M38-A protocol.Results All the strains were identified as classic A.fumigates,and fall into 11 CSP genotypes.The most common genotypes were t04A (n =32),t03 (n =17) and t01 (n =24) in all the strains,tl0,t04A and t01 in Fujian,t04A and t01 in Shanghai,t01,t03 and t04A in Hebei,t02,t04A,t01and t03 in Beijing.One A.fumigatus strain was identified as a new CSP type t25 in Fujian,which showed no obvious difference in morphology,growth rate or appropriate growth temperature from the other CSP genotypes of A.fumigatus strains.No statistical difference was found in the susceptibility to amphotericin B,itraconazole or fluconazole among different genotypes of A.fumigates,whereas the MICs of itraconazole were significantly lower in A.fumigates isolates from Fujian than in those from the other three regions.Conclusions The CSP genotypic profile of A.fumigates varies in clinical isolates from different regions.No significant difference is observed in the susceptibility to amphotericin B,itraconazole or fluconazole among different CSP genotypes of A.fumigates,but the susceptibility to itraconazole is somewhat different between A.fumigates strains from different regions.

Influence of Different Fixing Methods on Cardiac Index in C57 Mice of Wild Type andβ-adrenoceptor Knockout Mice / 世界科学技术-中医药现代化

This study was aimed to choose the best fixing method of electrocardiogram (ECG) measurement in mice by comparison of ECG index in three different fixing ways. Changes of ST amplitude and heart rate in ECG were measured in the self-made mouse sets, tape bundling or breathing anesthesia state. The amplitudes of the ST seg-ment in C57BL6 mice and β1/β2-AR double knockout mice were elevated in tape bundling or breathing anesthesia state in comparison with that in the self-made mouse sets (all P < 0.01). The heart rate under the tape bundling state in C57BL6 mice was significantly faster than that in the self-made mouse sets (P< 0.05 or P< 0.01). And the heart rate under anesthesia in β1/β2-AR double knockout mice was significantly slower than that in tape bundling state or the self-made mouse sets (both P < 0.01). It was concluded that relative to the binding and anesthesia method, fixing mouse in the self-made mouse sets, which can more truly reflect the physiological or pathological states, is a relatively good fixing method of ECG measurement in experimental study.

Investigation on Effects and Mechanism of Acupoints / 世界科学技术-中医药现代化

Acupoints are surface reaction and treatment spots under the pathological conditions which have functions of diagnosis and treatment. For the study of acupoints, we can carry it out from the aspects of structure and function. The two aspects have a close relationship. Structure is the material basis of function, while function is the manifesta-tion of structure. Therefore, the study of acupoints is not only significant to reveal the essence of acupoint, but also has enormous value in the guidance of acu-moxibustion for clinical practice. Staring from the normality of structure and variability of function, we explored the essence of acupoints, and further revealed ordered regulation rule and its biological basis, so that the essence of acupuncture points will be illustrated with scientific evidences.

Clinical Observation of Fire-needle Therapy plus Tuina for Senile Knee Osteoarthritis / 针灸推拿医学（英文版）

Objective: To observe the therapeutic efficacy of fire-needle therapy plus tuina in treating primary knee osteoarthritis (KOA) in the elderly. Methods: Twenty-three patients with KOA were intervened by pricking with fire-needle therapy plus tuina, and evaluated by using the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) before and after intervention. Results: After intervention, the total score, scores of pain, rigidity, and activities of daily living of WOMAC dropped significantly (P Conclusion: Pricking with fire needle plus tuina can produce a significant therapeutic efficacy in treating KOA in the elderly, thus worth application in clinic.

Rapid Detection and Identification of Pathogenic Fungi of Some Deep Fungal Infections by PCR in Combination with Genescan Analysis / 中华皮肤科杂志

Objective To rapidly detect and identify pathogenic fungi of some deep fungal infections by PCR.Methods The suspensions of22pathogenic fungi(23strains)were amplified by PCR with fungal universal primers ITS86and ITS4which were labeled by FAM.The precise length of amplified fragments was determined by ABI PRISM TM 377Sequencer and Genescan analysis software,then compared with that of am-plicons of corresponding fungal DNA which were previously extracted.Results(1)Amplification of17pathogenic fungi with ITS4,ITS86resulted in a unique fragment length(except for A.nidulans and A.niger,C.albicans and C.stellatoidea,F.pedrosoi and E.dermatitidis).(2)No significant difference of the length of am-plicons was found between the fungal suspension and control organisms,based on the results of Genescan analysis.(3)The whole process took only6h to complete the detection.Conclusion The combination of fun-gal suspension PCR with ITS fungal universal primers and Genescan analysis might provide an accurate,spe-cific,sensitive,and rapid approach to detect and identify22pathogenic fungi causing deep fungal infections,and hold promise to be applied for the diagnosis of deep fungal infection.

Expression of Secretory Aspartyl Proteinase in Human Vulvovaginal Candidiasis / 中华皮肤科杂志

Objective To investigate the differential expression of secretory aspartyl proteinase in patients with vaginal C.albicans infection and in asymptomatic candidal carriers.Methods Secretory aspartyl proteinase expression was determined with reverse transcription-PCR in vaginal specimens taken from 10 asymptomatic candidal carriers,14 patients with vulvovaginal candidiasis (VVC) and 10 patients with recurrent VVC (RVVC).Results SAP2 and SAP4 to SAP6 subfamily were detected in both candidal carriers and patients with vaginal candidiasis.SAP1 and SAP3 transcripts were not observed in 10 asymptomatic candidal carriers.All 9 SAP genes were differently expressed in VVC and RVVC patients.SAP1 and SAP3 transcripts were frequently amplified in some patients.Conclusion It is shown that C.albicans infection is associated with differential expression of individual SAP genes,which may be involved in the pathogenesis of vaginal candidiasis.

Expression of MCP-1 and MIP-2 mRNA in Vagina of Murine Model of Vulvovaginal Candidiasis / 中华皮肤科杂志

Objective To investigate the kinetic expression level of chemokines (MCP-1 and MIP-2) in vagina of a murine model of vulvovaginal candidiasis (VVC).Methods The estrogen-treated murine model of VVC was set up.Vaginal specimens were obtained in different duration after inoculation of C.albicans intravaginally.Semi-quantitative RT-PCR was applied to determine MCP-1 and MIP-2 mRNA levels in these tissues.Results Compared with the control mice treated with olive oil,persistent growth of C.albicans was found from the 2nd day to 21st day after inoculation in estrogen-treated mice.Significantly higher levels of MCP-1 mRNA were observed in vaginal tissues in infected estrogen-treated mice than those in other 2 groups,infected but non estrogen-treated mice and estrogen-treated but uninfected mice.The high level of MCP-1 mRNA maintained from the 4th day to 21st day in infected estrogen-treated mice.It was also found that levels of MIP-2 mRNA were significantly higher in the vagina in the 2nd day in 3 groups of mice than those in naive mice,however,no significant difference was shown among 3 groups throughout the study period.Conclusion High level of MCP-1,rather than MIP-2,may be associated with susceptibility to VVC.

Species and Genotypes of Clinical Isolates of Candida Strains from Patients with Vulvovaginal Candidiasis / 中华皮肤科杂志

Objective To investigate the species and DNA polymorphism of Candida strains from different individuals and to find the relationship between the species or DNA patterns of strains and different patient groups. Methods The present study chose the isolates from 3 different sources groups(from the patients with vulvovaginal candidiasis, recurrent vulvovaginal candidiasis or asymptomatic carriers respectively). Germ tube test, chlamydospore test, CHROMagar Candida and API20 kit system were applied to separate non-Candida albicans strains from Candida albicans. Random amplification of polymorphic DNA (RAPD) analysis was used to study DNA typing of 97 strains of Candida collected from different patients and to determine whether isolates from different patients were genetically similar or dissimilar. The data were analyzed by SPSS10.0. Results The ninety-seven isolates consisted of 83 strains of Candida albicans and 14 of non-Candida albicans. Of 9 random primers used, two primers (primer 4 and primer 7) gave good reactions, the sequences of which were 5′-ACCCGACCTG-3′, 5′-GGTGACGCAG-3′ respectively. The 99 isolates could be classified into 19 and 21 genotypes by the two primers respectively. Different genotype was not shown in most isolates from different groups. A particular genotype associated with different conditions was seen in only a few isolates. Conclusion Candida albicans is the main pathogenic yeasts and most strains of non-Candida albicans are C.glabrata in the vulvovaginal candidiasis. Genotyping of most isolates didn′t show obvious correlation with different patient groups.

Identification of Genetic Locus for Disseminated Superficial Actinic Porokeratosis / 中华皮肤科杂志

Objective To identify the genetic locus for disseminated superficial actinic porokeratosis(DSAP).Methods Genome DNA was extracted from the whole blood of the family members of a pedigree of DSAP.Genotyping on chromosome12q that had been identified was performed by using7microsatellite mark-ers to scan the family members of DSAP and analysed with LINKAGE(5.1Version).Results A maximum2-point lod score of5.15with marker D12S79at a recombination fraction(?)=0.00was found.Conclusion Our study supports that DSAP gene localizes at the long arm of chromosome12,which was first reported in the literature.

Evidence for Psoriasis susceptibility Locus in China:Locus on Chromosome 6p21.3 / 中华皮肤科杂志

Objective To elucidate the genetic base of psoriasis for Chinese patients, the positional candidate loci (D6S273?D6S276?D6S422?D6S299?D6S291?D4S1535?D4S1652?D4S171)previously reported in the regions 6p21.3 and 4q were studied in some carefully examined psoriatic families in order to establish whether the eight reported microsatellites loci(STRs) underlay susceptibility to psoriasis in different populations. Methods Two hundred and five probands with psoriasis vulgaris were identified from outpatients attending the Institute of Dermatology, Chinese Academy of Medical Sciences. Genotypes were generated at 8 polymorphic loci on chromosome 6p21 and 4q in 14 pedigrees. The results were analyzed parametrically by linkage 5.0 software. Results There was evidence for linkage to D6S273 in 6p21.3 (the LOD score was 1.26) . No evidence for linkage was obtained at other loci including three loci on chromosome 4q. Conclusions This study confirms the presence of a psoriasis susceptibility locus on chromosome 6p previously studied. It is shown that there may be psoriasis susceptibility locus D6S273 on chromosome 6p21.3 in the Chinese population.