Objective To observe the effects of electroacupuncture on intestinal flora and serum inflammatory factors in rabbit model with rheumatoid arthritis(RA);To explore the mechanism of its therapeutic effect on RA.Methods RA model rabbits were established by ovalbumin induction combined with Freund's complete adjuvant,and the rabbits after successful modeling were randomly divided into model group,probiotic group and electroacupuncture group,with 6 rabbits in each group.Another 6 rabbits were set as the normal group.The electroacupuncture group received electroacupuncture at the bilateral"Zusanli"and"Dubi"for 30 minutes,the probiotic group was given probiotic capsules solution(14.5 mg/kg)by gavage,once a day,for two consecutive weeks.The knee joint circumference and pain threshold of rabbits were measured,histopathological morphological changes of colonic tissue and synovial tissue ws observed by HE staining,16S rDNA sequencing was used to analyze structural changes of intestinal flora,the contents of TNF-α,IL-1β and IL-6 in serum were detected by ELISA.Results Compared with the normal group,the circumference of knee joint of rabbits in the model group increased significantly,the pain threshold was significantly decreased(P<0.01);the colonic mucosal damage was serious,the goblet cells were missing,a large number of inflammatory cells were infiltrate;the joint capsule synovial surface was rough,the synovial cell layer was hyperplasia and thickening,the synovial tissue inflammatory cell infiltration was obvious;the number and evenness of gut microbiota species decreased,while the relative abundance of Proteobacteria,Firmicutes,and Bacteroidetes decreased(P<0.05,P<0.01),the relative abundance of Desulfobacteria increased(P<0.01),while the relative abundance of Campylobacter,Lawsonella,and Pseudomonas increased(P<0.01),while the relative abundance of Heshanomonas and Herbaspirillum decreased(P<0.01);the contents of serum TNF-α,IL-1β and IL-6 increased(P<0.01).Compared with the model group,the knee joint circumference of the probiotic group and the electroacupuncture group decreased,the pain threshold increased(P<0.01,P<0.05);the degree of intestinal mucosal damage was reduced,the goblet cells were basically arranged neatly,the inflammatory cell infiltration was reduced;synovial cells proliferation and thickening decreased,with a small amount of inflammatory cell infiltration;the number and evenness of gut microbiota species increased,while the relative abundance of Proteobacteria,Firmicutes,and Bacteroidetes increased(P<0.05,P<0.01),the relative abundance of Desulfobacteria decreased(P<0.01),while the relative abundance of Campylobacter,Lawsonella and Pseudomonas decreased(P<0.01),the relative abundance of Heshanomonas and Herbaspirillum increased(P<0.05,P<0.01);the contents of serum TNF-α,IL-1β and IL-6 significantly decreased(P<0.01).Conclusion Electroacupuncture can significantly improve the symptoms of RA rabbits and reduce the inflammatory reaction in synovial tissue of joint,and the mechanism may be related to the regulation of intestinal flora structure.

Humans ; East Asian People ; Surveys and Questionnaires ; Vitiligo/epidemiology*

BACKGROUND: This study aimed to develop a comprehensive instrument for evaluating and ranking clinical practice guidelines, named Scientific, Transparent and Applicable Rankings tool (STAR), and test its reliability, validity, and usability. METHODS: This study set up a multidisciplinary working group including guideline methodologists, statisticians, journal editors, clinicians, and other experts. Scoping review, Delphi methods, and hierarchical analysis were used to develop the STAR tool. We evaluated the instrument's intrinsic and interrater reliability, content and criterion validity, and usability. RESULTS: STAR contained 39 items grouped into 11 domains. The mean intrinsic reliability of the domains, indicated by Cronbach's α coefficient, was 0.588 (95% confidence interval [CI]: 0.414, 0.762). Interrater reliability as assessed with Cohen's kappa coefficient was 0.774 (95% CI: 0.740, 0.807) for methodological evaluators and 0.618 (95% CI: 0.587, 0.648) for clinical evaluators. The overall content validity index was 0.905. Pearson's r correlation for criterion validity was 0.885 (95% CI: 0.804, 0.932). The mean usability score of the items was 4.6 and the median time spent to evaluate each guideline was 20 min. CONCLUSION The instrument performed well in terms of reliability, validity, and efficiency, and can be used for comprehensively evaluating and ranking guidelines.
Reproducibility of Results ; Surveys and Questionnaires ; Practice Guidelines as Topic ; Humans

Gallbladder carcinoma is the most common malignant tumor of the biliary tract system. It is characterized by atypical early clinical manifestations, rapid progress and poor prognosis. There is no specific marker for gallbladder cancer, and its preoperative diagnosis mainly relies on imaging examination, while pet-ct is more advantageous in detecting the metastasis of locally advanced gallbladder cancer. Surgical treatment is still the main treatment, but most patients have lost the opportunity of surgical resection by the time of treatment. In recent years, there have been a lot of researches on the diagnosis and treatment of gallbladder cancer at home and abroad. This paper reviews the diagnosis and treatment of gallbladder carcinoma.

Objective: To investigate the timing of the laparoscopic cholecystectomy after percutaneous transhepatic gallbladder drainage for acute pyogenic cholecystitis in the advanced age. Methods: The clinical data for 56 advanced age patients with acute pyogenic cholecystitis in the Department of Hepatobiliary Surgery, the 900th Hospital of the Joint Logistics Support Force of People′s Liberation Army from January 2018 to February 2019 were retrospectively analyzed. There were 31 males and 25 females, aged from 70 to 86 years, with average age was (75.52±3.57) years. According to the percutaneous transhepatic gallbladder drainage(PTGD) after laparoscopic cholecystectomy(LC) time interval, all patients were divided into three groups. Patients in the group A(n=12), B(n=21), and C (n=23) performed LC were within 2 months, during 2-4 months, and during 4-6 months, retrospectively. Observation indicators: (1) Surgical situations. The operation and postoperative basic condition of the three groups were compared. (2) Follow-up situations. Patients were followed-up by outpatient examination or telephone interview to detect the postoperative complication in the postoperative three months up to June 2019. Measurement data with normal distribution were represented as (Mean±SD), and comparison multiple groups was done using single factor analysis of variance (AVONA test) , and comparison between groups was done using the t test, and comparison of multiple groups in pairs was done using the SNK-q test, and hierarchical data were analyzed using Kruskal-wallis H test. Count data were analyzed using the chi-square test or Fisher exact probability. Results: (1) Surgical situations: the thickness of gallbladder wall before LC, the rates of converting to laparotomy, the volume of intraoperative blood loss, the operation duration, and the duration of postoperative hospital stay were (0.57±0.04) cm, 50.0%, (95.83±11.45) ml, (107.50±21.90) min, (5.67±3.40) d in the group A, and (0.43±0.03) cm, 9.5%, (69.52±24.59) ml, (71.43±12.16) min, (3.76±2.61) d in the group B, and (0.43±0.05) cm, 39.1%, (68.64±21.89) ml, (77.95±12.88) min, (5.05±2.95) d in the group C, respectively, showing significant differences in the above indicators between the three groups (P<0.05). The thickness of gallbladder wall before LC, the volume of intraoperative blood loss, the operation duration, those in group A were higher than the group B and C (P<0.05), and with no statistically significant different between the group B and C (P>0.05). The rates of converting to laparotomy, the duration of postoperative hospital stay in group B were better than the group A and C (P<0.05), and with no statistically significant different between the group A and C (P>0.05). The thickness of gallbladder wall before LC, the volume of intraoperative blood loss, the operation duration, and the duration of postoperative hospital stay were (0.43±0.03) cm, (46.67±9.82) ml, (67.69±7.77) min, (2.64±0.58) d in the gallbladder wall thickness of successful LC patients, and (0.52±0.04) cm, (123.53±17.30) ml, (134.12±25.51) min, (8.47±0.80) d in the laparotomy patients, respectively, showing significant differences in the above indicators between the two groups (P<0.05). (2) Follow-up situations: 56 patients were followed up and without perioperative death. No complications occurred after 3 months of follow-up. Conclusion Elective surgery that is performed in 2-4 months after PTGD for patients with acute pyogenic cholecystitis in the advanced age can reduce the volume of intraoperative blood loss and the rates of converting to laparotomy, shorten the operation duration and the duration of postoperative hospital stay, which is beneficial to the recovery of patients.

OBJECTIVETo compare the effects of different acupuncture methods on urine metabolites in rheumatoid arthritis (RA) rabbits, and to explore the specificity mechanism of heat-reinforcing acupuncture for RA.

METHODSA total of 40 clean purple-blue rabbits were randomly allocated to a normal group, a model group, a mild reinforcing-reducing needling (MRRN) group, a twirling-reinforcing needling (TRN) group and a heat-reinforcing needling (HRN) group, 8 rabbits in each one. Except the normal group, the rabbits in the remaining groups were treated with ovalbumin and freezing to establish RA model. The rabbits in the MRRN group, TRN group and HRN group were treated with MRRN, TRN and HRN at "Zusanli" (ST 36), respectively, 30 min per treatment, once a day for seven days. After treatment, 24-h urine was collected. The rabbits were sacrificed to collect synovial tissues of knee to perform morphology observation; the liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q/TOF-MS) was applied to measure urine metabolites. All the data were analyzed by principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA).

RESULTSCompared with the normal group, the leucine-related metabolites, as main urine metabolites, were decreased in the model group (<0.05), while the purine-related metabolites and tryptophane-related metabolites were increased (<0.05). Compared with the model group, the leucine-related metabolites, as main urine metabolites, were increased in the three needling groups after treatment (<0.05), while the tryptophan-related metabolites andpurine-related metabolites were decreased (<0.05), moreover, the leucine-related metabolites in the HRN group were obviously higher than those in the MRRN group and TRN gruop (<0.05).

CONCLUSIONSMRRN, TRN and HRN can regulate the pathway of leucine metabolism (energy metabolism), purine metabolism (oxidative damage) and tryptophane metabolism (immune regulation) for RA, The specificity of HRN for RA focuses on regulation of leucine metabolism (energy metabolism).

OBJECTIVETo explore heat-reinforcing needling for the metabolite profiling changes in serum of rheumatoid arthritis (RA) rabbits with liquid chromatograph-mass spectrometer (LC-MS) technique, and to investigate its mechanisms.

METHODSForty clean purple blue rabbits were randomized into a normal group, a model group, a reinforcing-reducing needling (RRN) group, a twirling-reinforcing needling (TRN) group, and a heat-reinforcing needling (HRN) group, 8 cases in each group. RA rabbits with cold syndrome were made with ovalbumin and freezing except those in the normal group. No treatment was given in the normal and model groups. The corresponding manipulations for 7 days were applied at "Zusanli" (ST 36) in the three acupuncture groups, 30 min a time, once a day. After intervention the pain threshold and the local skin temperature of each group were observed. Fresh serum from heart was collected for metabonomics detection. Principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) were adopted. Several metabolites were screened by the variable importance in the projection values (VIP>1) andvalue (<0.05).

RESULTSThe pain threshold and the local skin temperature in the model group were lower than those in the normal group (both<0.05). The pain threshold and the local skin temperature in the three acupuncture groups were higher than those in the model group after intervention (all<0.05), which were better in the HRN group than those in the RRN and TRN groups (all<0.05). The serum metabolites of carnitine, LysoPC (14∶0), LysoPC (18∶3), LysoPE (0∶0/20∶5), LysoPE (0∶0/22∶1), decylic acid, stearic acid and lactic acid in the model group increased compared with those in the normal group, and other metabolites decreased, including leucine, valine, glutamine, pyroglutamic acid, α-ketoglutaric acid, succinic acid, fumaric acid, malic acid, galactose, mannose. Those metabolites were correlated fatty acid, amino acid, citric acid cycle, and glucose metabolism. The metabolites above-mentioned in the three acupuncture groups were regulated in various degrees (all<0.05). Lactic acid decreased and succinic acid, fumaric acid, malic acid, galactose, mannose increased more obviously in the HRN group than those in the RRN and TRN groups.

CONCLUSIONThe specificity of heat-reinforcing needling for RA presents the regulation for citric acid cycle and glucose metabolism.

OBJECTIVETo compare the efficacy differences between heat-reinforcing needling and conventional treatment of western medicine on Kashin-Beck disease (KBD) with cold-dampness blocking collaterals syndrome.

METHODSSixty KBD patients of cold-dampness blocking collaterals syndrome were randomly assigned into a heat-reinforcing needling group and a western medication group, 30 cases in each one. In the heat-reinforcing needling group, the heat-reinforcing needling was applied at local painful sites, combined with the acupoints based on the syndrome differentiation and the distal acupoints on the affected meridians. Acupuncture was given 30 min per time, once a day, the treatment of 5 days made 1 session; there was an interval of 2 days between two sessions. In the western medication group, sodium selenite tablets were prescribed for oral administration after meals, 2 tablets each time, once a day; ibuprofen sustained release capsules were prescribed for oral administration, 1 capsule each time, twice a day; vitamin C tablets were prescribed for oral administration, 2 tablets each time, three times a day. Four-week treatment was given in the two groups. The Western Ontaraio and Mcmaster Universities Osteoarthritis Index (WOMAC) was adopted to assess the involved joints; the safety was assessed in the process of treatment; the efficacy was analyzed, and the follow-up visit was conducted 3 months and 6 months after treatment, respectively.

RESULTSAfter 4-week treatment, the total effective rate was 96.7%(29/30) in the western medication group, which was superior to 90.0% (27/30) in the heat-reinforcing needling group (<0.05). However, the safety in the heat-reinforcing needling group was superior to that in the western medication group (<0.05). The improvements of joint function in 3-month and 6-month follow-up visits in heat-reinforcing needling group were superior to those in western medication group (both<0.05).

CONCLUSIONSThe heat-reinforcing needling for KBD is safe and effective with less adverse reactions. The short-term effect of heat-reinforcing needling isinferior to western medication, but the long-term efficacy is remarkably superior to western medication.

Objective To measure the endogenous metabolites in synovial tissues of rheumatoid arthritis (RA) rabbits with cold syndrome;To investigate the specificity mechanisms of heat-reinforcing needling for RA. Methods A total of forty healthy purple blue rabbits were randomly allocated to normal group, model group, reinforcing-reducing needling group (RRN), twirling-reinforcing needling group (TRN) and heat-reinforcing needling group (HRN) 8 rabbits in each group. Arthritis rabbits cold model was made with ovalbumin and freezing. Except for normal groupand model group, RRN was given acupuncture of reinforcing-reducing needling at Zusanli (ST36), TRN was given acupuncture of twirling-reinforcing needling at Zusanli (ST36), HRN was administrated acupuncture of heat-reinforcing needling at Zusanli (ST36), once a day and retaining 30 min, a total of seven days. Fresh synovial tissues of rabbits knee joints were extracted after the intervention, then GC-Q/TOF-MS technology were employed to evaluate metabolic profiles. Results The synovial tissues metabolites of TCA cycle, carbohydrate and fatty acid metabolism in model group mainly increased compared with normal group (P<0.05); The synovial tissues metabolites of various kinds above-mentioned decreased compared with model group in all intervention of acupuncture group (P<0.05), and the metabolites of TCA cycle and carbohydrate metabolism in HRN were obviously less than that of RRN and TRN (P<0.05). Conclusion The specificity of heat-reinforcing needling for RA manifests in regulation of TCA cycle and carbohydrate metabolism.

Objective To observe the effect of costimulatory molecule B7-1 on cytoskeleton rearrangement in mouse podocytes induced by angiotensin Ⅱ (Ang Ⅱ),and to study the underlying molecular mechanism of B7-1 in the pathological changes of podocytes.Methods All cultivation of conditionally immortalized mouse podocytes (MPC) in vitro were divided into the following groups:normal control group,CTLA-4 group,Ang Ⅱ group (10-6 mmol/L 12 h,24 h;10-8 mmol/L 12 h,24 h) and CTLA-4 with Ang Ⅱ group.Transfect B7-1 RNA interference fragment (siRNA) to the mature podocytes,and then restimulated by Ang Ⅱ (10-6 mmol/L 12 h),the change of podocyte cytoskeleton after Ang Ⅱ stimulation were observed.The expression of B7-1 in each group was assayed by flow cytometry and Western blotting.The nephrin and p-nephrin protein levels in the four groups were also analyzed by Western blotting.At the same time,the podocyte cytoskeleton distribution as indicated by F-actin was observed by fluorescence microscopy.Results Flow cytometry and Western blotting showed that B7-1 was not expressed in the normal control group.Ang Ⅱ showed a concentration and time dependent induction of B7-1 expression in mouse podocytes (P ＜ 0.05).Western blotting indicated that Ang Ⅱ induced B7-1 protein expression (P ＜ 0.05).Expression of nephrin and p-nephrin was significantly down-regulated by Ang Ⅱ (P ＜ 0.05).Compared with the normal control group,the expression of podocyte protein nephrin and p-nephrin in Ang Ⅱ stimulation group was significantly reduced (P ＜ 0.05).Using FITC phalloidin fluorescence staining showed that CTLA-4+Ang Ⅱ stimulation group cytoskeleton rearrangement was improved significantly and F-actin recombinant score (mCFS) decreased compared with Ang Ⅱ group (P ＜ 0.05),suggesting that Ang Ⅱ led to the disorder of the podocytes cytoskeleton and the destruction of the cytoskeleton of podocytes by Ang Ⅱ could be improved after B7-1 blocking.Compared with the Ang Ⅱ stimulation,transfection of B7-1 siRNA + Ang Ⅱ stimulation group improved F-actin cytoskeletal rearrangement,and mCFS also decreased significantly (P ＜ 0.05),suggesting that transfection of B7-1 siRNA might improve the damage of Ang Ⅱ on podocytes cytoskeleton.Conclusions B7-1 participates in the process of cytoskeleton reconstruction and plays an important role in the pathological changes of podocytes.