1.Effect of Huanglian Ejiaotang on 5-Hydroxytryptamine System and Gut Microbiota in Sleep-deprived Rats
Huaqiong DIAO ; Dan WEI ; Haiyue DING ; Jing ZHANG ; Yufei CHEN ; Min WANG ; Qingsheng ZHU ; Xiaoli LI
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(21):49-58
ObjectiveTo explore the mechanism of Huanglian Ejiaotang in intervening in insomnia based on 5-hydroxytryptamine (5-HT) system and gut microbiota. MethodFifty-five SPF-grade SD rats were randomly divided into normal group, model group, low-, medium-, and high-dose Huanglian Ejiaotang groups (1.925, 3.85, and 7.7 g·kg-1), and Estazolam group (0.1 mg·kg-1). Except for those in the normal group, the rats in the other five groups were subjected to sleep deprivation on a narrow platform for 12 hours daily for 21 consecutive days. After 14 days of drug intervention, the sleep, exploratory behavior, and depressive-like behavior of the rats were assessed using the pentobarbital sodium sleep synergistic test, the open field test, and the sugar preference test, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of 5-HT, 5-hydroxyindoleacetic acid (5-HIAA), tryptophan hydroxylase (TPH), and monoamine oxidase-A (MAO-A). Real-time polymerase chain reaction (Real-time PCR) was used to measure the mRNA expression of the 5-HT1A receptor (5-HT1AR) and 5-HT2A receptor (5-HT2AR). Differences in gut microbiota among the groups were assessed using 16S rRNA sequencing, and the correlation between the 5-HT system and microbiota was revealed using redundancy analysis. ResultCompared with the normal group, the model group showed a prolonged sleep latency (P<0.05), reduced sleep maintenance (P<0.01), decreased central area activity time in the open field (P<0.01), and reduced sugar preference rate (P<0.05). Moreover, the model group also showed decreased levels of 5-HT, 5-HIAA, TPH, and MAO-A (P<0.01), decreased 5-HIAA/5-HT ratio (P<0.01), downregulated mRNA expression of 5-HT1AR (P<0.01), and upregulated mRNA expression of 5-HT2AR (P<0.05). The proportion of Firmicutes decreased, while that of Bacteroidetes increased, leading to a decreased Firmicutes/Bacteroidetes (F/B) ratio (P<0.05). Compared with the model group, the high-dose Huanglian Ejiaotang group exhibited a shortened sleep latency (P<0.01), and increased sleep maintenance (P<0.01). The low-dose Huanglian Ejiaotang group showed increased central area activity time (P<0.01) and an increased sugar preference rate (P<0.05). The high-dose Huanglian Ejiaotang group exhibited increased levels of 5-HT, 5-HIAA, TPH, and MAO-A (P<0.01), increased 5-HIAA/5-HT ratio (P<0.05), upregulated mRNA expression of 5-HT1AR (P<0.01), and downregulated mRNA expression of 5-HT2AR (P<0.05). The low-dose Huanglian Ejiaotang group displayed an increased proportion of Firmicutes and a decreased proportion of Bacteroidetes, resulting in an increased F/B ratio. At the phylum level, 5-HT, 5-HIAA, and MAO-A were positively correlated with Firmicutes and negatively correlated with Bacteroidetes. At the genus level, 5-HT, 5-HIAA, TPH, and MAO-A were negatively correlated with Prevotella and Lactobacillus and positively correlated with Blautia and Bacteroides. ConclusionHuanglian Ejiaotang can improve sleep deprivation-induced insomnia and depressive-like behavior by regulating the activity of the 5-HT system and the composition of gut microbiota.
2.Indocyanine green fluorescence navigation in obese patients undergoing laparoscopic cholecystectomy
Yongzhen ZHOU ; Qingsheng FU ; Tao LI ; Xudong ZHANG ; Chunfu ZHU ; Xihu QIN ; Baoqiang WU
International Journal of Surgery 2023;50(12):846-851
Objective:To investigate the feasibility and potential of fluorescent cholecystic bile duct visualization with direct intravenous injection of indocyanine green(ICG) in obese patients undergoing laparoscopic cholecystectomy(LC).Methods:The clinical data of 132 patients with LC combined with obesity admitted to the Department of Hepatobiliary and Pancreatic Surgery of the Second People′s Hospital of Changzhou City, affiliated to Nanjing Medical University, from January 2023 to July 2023 were retrospectively analyzed. They were divided into fluorescence group( n=65) and control group( n=67) according to whether indocyanine green fluorescence navigation was used or not. There were 50 males and 82 females, and all the enrolled patients body mass index≥28 kg/m 2. Two groups identify the time of the three tubes, intraoperative bleeding, operation time, postoperative hospitalization time, white blood cell count(WBC), C-reactive protein(CRP), alanine aminotransferase(ALT), gamma-glutamyl transferase(GGT), and postoperative follow-up in the fluorescent and control groups were counted respectively. Measurement data with skewed distribution were expressed as M( Q1, Q3), and intergroup comparisons were performed using the Mann-Whitney U test; counting data were described by frequency(rate), and intergroup comparisons were made by applying the chi-square test, Fisher′s exact probability method, and chi-square corrected test according to the difference in the minimum frequency. Results:Surgery was successfully completed in both groups. Preoperative inflammatory indicators and liver function levels were also not statistically significant( P>0.05). The time to identify the three tubes, operation time, intraoperative bleeding, and postoperative hospitalization in the fluorescence group were 18.00(13.50, 20.00) min, 40.00(30.00, 50.00) min, 5.00(5.00, 10.00) mL, and 2.00(1.50, 3.00) d, and in the control group were 32.00(25.00, 45.00) min, 65.00(50.00, 85.00) min, 41.00(41.00, 46.00) mL, and 4.00(3.00, 5.00) d. The differences between the two groups were statistically significant( P<0.05). The postoperative leukocyte count, postoperative CRP, and postoperative GGT were 9.15(7.10, 11.75)×10 9/L, 7.19(3.22, 20.00) mg/L, and 34.0(20.0, 49.0) U/L in the fluorescence group, and 13.05(11.02, 15.59)×10 9/L, and 18.78(12.90, 32.95) mg/L in the control group, respectively, 82.5(68.5, 114.5) U/L, and the differences between the two groups were statistically significant ( P<0.05). None of the patients showed abdominal pain, abnormal liver function and hepatobiliary ultrasound in the follow-up findings within 2 months after surgery. Conclusion:The effect of obesity, a factor that interferes with ICG fluorescence, is extremely limited, and ICG fluorescence cholangiography is a useful technique in the obese population that not only improves the efficiency of the procedure, but also increases intraoperative safety, with results superior to those of conventional laparoscopic cholecystectomy.
3.Coxiella burnetii in periprosthetic joint infection: a case and systematic review
Chongfei YANG ; Shu ZHU ; Feike KANG ; Xiaorui CAO ; Qingsheng ZHU
Chinese Journal of Orthopaedics 2023;43(3):185-190
One case of periprosthetic infection after artificial joint replacement caused by Coxiella burnetii was reported. The patient was admitted to hospital due to joint pain and sinus formation after artificial knee replacement. Through medical history, physical examination, imaging examination and gene detection, it was confirmed that the infection around the prosthesis was caused by Coxiella burnetii. The patient was treated with two-stage revision surgery combined with sensitive antibiotics. Through literature review, a total of 9 cases of Coxiella burnetii infection after artificial joint replacement were reported, including 6 cases of hip joint and 3 cases of knee joint; 4 cases had clear pathogen exposure history; only one case was found protheses loosening on imaging; 7 cases with positive IgG antibody against Coxiella burnetii; 7 cases were positive for gene detection; 8 cases were negative in etiological examination, and 1 case was not cultured. All patients were treated with doxycycline and hydroxychloroquine. The serological index of 1 case without Q fever symptom turned negative after treatment. 1 case with multiple system organ dysfunction relieved symptoms after treatment, and serological antibody indicators and nuclear medicine results turned negative. All 7 patients were cured after secondary revision surgery. Coxiella burnetii infection after total joint arthroplasty is rare and lacks typical imaging manifestations and clinical features. The diagnosis of Coxiella burnetii infection after total joint arthroplasty mainly depends on gene detection and serological detection. Two-stage revision surgery combined with sufficient and long-term sensitive antibiotics is the main method of treatment.
4.Application of intraoperative intravenous injection of indocyanine green in endoscopic surgery for chronic atrophic cholecystitis
Qingsheng FU ; Lei JIN ; Tao LI ; Xudong ZHANG ; Chunfu ZHU ; Xihu QIN ; Baoqiang WU
International Journal of Surgery 2022;49(1):5-10,F3
Objective:To investigate the clinical value of intraoperative intravenous injection of indocyanine green in differentiating extrahepatic bile duct structure in chronic atrophic cholecystitis.Methods:A retrospective analysis was performed on the data of 110 patients diagnosed with chronic AC who underwent laparoscopic cholecystectomy (LC) admitted to the Department of Hepatobiliary and pancreatic Surgery of Changzhou Second People′s Hospital from January 2020 to July 2021. All patients were confirmed by abdominal B-ultrasound before surgery. The patients in the experimental group were divided into experimental group ( n=55) and control group ( n=55) according to whether indocyanine green was intravenously injected during the operation. The experimental group was intravenously injected with 5 mg indocyanine green during the operation, and LC was navigated by indocyanine green fluorescence imaging technique during the operation. The control group received routine LC. The imaging rate and imaging time of the cystic duct, common bile duct, and common hepatic duct in the experimental group were compared. The clinical data, identify three tube time, operation time, intraoperative blood loss, abdominal cavity drainage placement and extubation time, transfer laparotomy and bile duct injury, postoperative hospital stay, postoperative first review of alanine aminotransferase (ALT), glutamine transferase (GGT) of leveling and follow-up were compared between the two groups. The measurement data subject to normal distribution were expressed by Mean±standard deviation ( ± s), and the two groups were compared by independent sample t test. The measurement data of skewness distribution were described by M( Q1, Q3)and the manhui method in nonparametric test was used Mann-whitney U test.The chi-square test or Fisher′s exact probability method was used for comparison between groups of count data. Results:Operation was performed successfully in both groups. In the experimental group, the common hepatic duct, common bile duct and gallbladder duct were developed successfully in all patients, 54 cases and 52 cases respectively, and the developing time of the three tubes was (15.8±1.2) min. In the experimental group, the time of three tubes, operation time, intraoperative blood loss and abdominal drainage tube placement were (18.5±1.3) min, (64.0±6.8) min, (16.3±6.7) mL, 43 cases, respectively. In the control group, there were (46.3±8.1) min, (98.7±10.5) min, (53.6±14.9) mL and 55 cases, respectively. The experimental group was significantly lower than the control group, and the difference between the two groups was statistically significant ( P< 0.05). There was no case of conversion to laparotomy and bile duct injury in the experimental group, and 1 case of conversion to laparotomy and 1 case of bile duct injury in the control group, and there was no statistical significance between the two groups ( P>0.05). There were significant differences in postoperative extubation time and postoperative hospital stay between the two groups ( P<0.05). ALT and GGT levels were 47(31, 75) U/L and 38(19, 114) U/L in the experimental group and 62(53, 92) U/L and 76(63, 96) U/L in the control group at the first postoperative review, with statistically significant differences between the two groups ( P<0.05). Patients in both groups were followed up for 3 months after discharge. There were no obvious complications in the experimental group, and 1 case had a small amount of peritoneal effusion 7 days after discharge in the control group. Conclusion:In the face of LC with chronic AC, intraoperative intravenous injection of indocyanine green to develop extrahepatic bile duct can help to distinguish its anatomical structure and avoid bile duct injury, improve the safety and progress of surgery, and maximize training and improve the level of the surgeon.
5.Evaluation of the efficacy of transgallbladder injection of indocyanine green in symptomatic gallbladder stones combined with liver cirrhosis surgery
Qingsheng FU ; Yongzhen ZHOU ; Tao LI ; Xudong ZHANG ; Lei JIN ; Chunfu ZHU ; Xihu QIN ; Baoqiang WU
International Journal of Surgery 2022;49(8):532-538,F3
Objective:To investigate the feasibility and efficacy of transcystic injection of indocyanine green during laparoscopic cholecystectomy (LC) surgery in the treatment of patients with gallbladder stones combined with liver cirrhosis.Methods:The clinical data of 96 patients with cirrhosis who underwent LC for gallbladder stones with cholecystitis attacks in the Department of Hepatobiliary and Pancreatic Surgery of the Second People′s Hospital of Changzhou City from January 2018 to May 2022 were retrospectively analyzed. All patients were diagnosed by clinical history and auxiliary examination before surgery and underwent cholecystectomy according to their groups, which were divided into fluorescence group ( n=49) and white light group ( n=47) according to whether ICG was used or not, where the fluorescence group underwent LC in fluorescence mode after direct intraoperative injection of ICG via gallbladder. The two groups were compared in terms of identification time of the three tubes, operation time, intraoperative bleeding, intraoperative injury, intraoperative open abdomen and blood transfusion, postoperative drainage time, postoperative hospitalization time, postoperative complications and changes in infection and liver function indexes before and after surgery. The measurement data obeying normal distribution were expressed as mean±standard deviation( ± s), and independent sample t-test was used for comparison between groups. The measurement data obeying the skewed distribution were expressed by M( Q1, Q3), and the Mann-Whitney U test was used for comparison between groups. Count data were expressed as rates (%), and the chi-square test or Fisher′s exact probability method was used for comparison between groups. Results:The procedure was successfully performed in both groups, and the identification of triple-tube time, operative time, intraoperative bleeding, postoperative drainage time, postoperative hospital stay, postoperative ALT, postoperative GGT, and postoperative TBIL in the fluorescent group were (14.96±1.49) min, (52.14±7.36) min, 5(5, 10) mL, (1.61±0.61) d, (2.37±0.49) d, 31.5(22.0, 44.8) U/L, 38.0(21.0, 91.5) U/L, 18.0(11.5, 22.8) μmol/L, and (29.87±3.37) min, (84.36±13.25) min, 10(10, 20) mL, (2.70±0.69) d, (3.15±0.42) d, 45.0(28.0, 64.8) U/L, 73.0(32.0, 132.0) U/L, 23.0(16.1, 29.3) μmol/L in the white light group and the differences were statistically significant compared with the two groups( P<0.05). In the fluorescence group, there was no intraoperative injury and no cases of intraoperative opening, and there were 2 cases of postoperative complications, while in the white light group, there were 2 cases of intraoperative injury, 1 case of intraoperative opening, and 5 cases of postoperative complications, and there were no cases of blood transfusion in both groups. There was no statistically significant difference between the fluorescence group and the white light group when comparing the preoperative laboratory indexes of both groups ( P>0.05). When comparing the first postoperative white blood cell count, C-reactive protein, aspartate aminotransferase, and alkaline phosphatase indexes in the fluorescence group and the white light group, there was no statistically significant difference between the two groups ( P>0.05). Conclusion:When LC is performed in patients with symptomatic gallbladder stones combined with cirrhosis, intraoperative injection of indocyanine green via the gallbladder to visualize the gallbladder and bile duct structures is simple and easy to perform, and the safe and efficient dissection of extrahepatic bile ducts and gallbladder bed speeds up the procedure while reducing postoperative trauma.
6.Research progress on time and dose selection of indocyanine green in laparoscopic cholecystectomy
Qingsheng FU ; Tao LI ; Xudong ZHANG ; Chunfu ZHU ; Xihu QIN ; Baoqiang WU
International Journal of Surgery 2021;48(7):472-477,F3
In the era of precision medicine, compared with open surgery, laparoscopic cholecystectomy has the advantages of less trauma, less postoperative complications and shorter hospital stay, but it cannot escape the risk of bile duct injury, and bile duct injury can occur at any time for various reasons, and its unpredictability greatly increases the difficulty of surgery. Although traditional cholangiography can avoid bile duct injury, it has not been widely used due to its limitations such as time consuming, high cost, high radiation and so on. Early indocyanine green has been widely used in lymph node examination, angiography, tumor localization and treatment due to its characteristics. Now, indocyane green fluorescence imaging technology has been gradually applied in LC, which can dynamically reflect the intraoperative situation and realize real-time surgical navigation, so it has achieved ideal effects in the identification of duct structure and prevention of bile duct injury. Indocyanine green injection preoperatively or intraoperatively through the gallbladder to develop extrahepatic bile ducts significantly improves the safety rate of surgery, but there is no unified standard for the use time and dosage of inddocyanine green. In this paper, the time and dose selection and clinical value of indocyanine green were reviewed based on the current research status and the author's experience.
7.A meta-analysis of the association between Caipain-10 gene polymorphism and T2DM in Chinese population
Chinese Journal of Diabetes 2018;26(1):24-31
Objective To evaluate the association between calpain-10 gene polymorphism and type 2 diabetes mellitus(T2DM) by system assessment method.Methods CNKI,CBM and WangFang Data were searched for studies assessing the genetic association between calpain-10 gene polymorphism and T2DM.The data from included studies were extracted to calculate for odds ratio (OR).The simplified version of STREGA list was used to evaluate the quality of researches.Review Manger 5.3.1 software was used to analyze the heterogeneity (I2 values) among the studies,α=0.05.Results A total of 21 randomized controlled studies were analyzed.3062 T2DM patients were involved in SNP43 loci polymorphism evaluation.The highest score of STREGA list was 6 points and the lowest score was 3 points.The proportions of 6 points,5 points,4 points and 3 points were 38.10%,33.33%,9.52% and 19.05%,respectively.Both scores of "whether having description of genotyping method" item and "whether having sufficient data" item were 0 point.Meta analysis results showed that SNP 43 mutation type "A" of Calpain-10 gene was related to T2DM (OR 0.60,95%CI 0.52,0.68,P<0.05).The ORs of SNP 19,44 and 63 for T2DM were 0.92,1.50 and 0.84,with P values 0.50,0.11 and 0.08,respectively.The sensitivity analysis showed the similar results,with a small publication bias.Conclusion SNP43 loci of Calpain-10 gene has a genetic association with T2DM.
8.Analysis of Changing the Original Test Result after Retesting in National Medical Device Sampling and Testing from 2013 to 2016.
Qing HAO ; Xintao ZHANG ; Xiao LI ; Jiong ZHU ; Qingsheng ZHANG ; Xian SHI
Chinese Journal of Medical Instrumentation 2018;42(1):56-57
The types and the reasons of changing the original test result after retesting in national medical device sampling and testing from 2013 to 2016, are summarized and analyzed. Firstly, collecting data of "standards not complied" and retesting. Then, giving specific examples when summarizing five types of changing the original test result after retesting. Meanwhile, analyzing the relevant reasons, discovering the deep problems. Finally, giving suggestions for the above problems.
Equipment and Supplies
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standards
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Quality Control
9.Expression of TLR4 in monocytes and its correlation with TNF-α and IL-6 in patients with diabetic neuropathy
Chinese Journal of Diabetes 2017;25(2):97-102
Objective To investigate the expression of Toll-like receptor 4 (TLR4)in peripheral blood monocytes and its correlation with TNF-α and IL-6 in type 2 diabetes (T2DM)patients with neuropathy. Methods Healthy volunteers (NC,n=44),T2DM without neuropathy (n=44)and T2DM with neuropathy (n=44)were recruited in the present study.The expression of TLR4 mRNA and protein in peripheral blood monocytes was detected by RT-PCR and flow cytometry. ELISA was undertaken to measure the plasma concentration of TNF-αand IL-6. Results The levels of HbA1 c[(9.09±1.62)%vs(8.36±1.10)% vs (5.30±0.89)%],the protein expression of TLR4[(42.02±9.69)% vs (31.27± 6.87)% vs (11.96±5.54)%],the mRNA expression of TLR4[(2.98±1.06)vs (1.74±0.47)vs (1.12 ±0.52)],TNF-α[(8.75±3.14 )vs (6.27±3.64)vs(3.19±1.17)pg/ml]and IL-6[(3.63±1.81)vs(2.60 ±1.14)vs(1.54±0.58)pg/ml]in patients with diabetic neuropathy significantly increased compared with T2DM and NC group;TLR4 showed a positive correlation with TNF-αand IL-6(r=0.631,P <0.0001;r=0.447,P =0.0023). Conclusion TLR4 of monocytes may take part in the occurrence of systemic inflammation and has a close relationship with the development of diabetic neuropathy.
10.Role of Mac-1 in osteoclast differentiation
Guoxi YANG ; Qingsheng ZHU ; Chongfei YANG
Chinese Journal of Pathophysiology 2017;33(3):539-542,547
AIM:To investigate the function of macrophage differentiation antigen-1 (Mac-1) in receptor acti-vator of nuclear factor-κB ligand ( RANKL )-induced osteoclast differentiation and the mechanisms .METHODS: The spleen cells were isolated from 4-week-old C57BL/6J mice, and cultured with RANKL, macrophage colony-stimulating fac-tor and CD11b and CD18 antibodies for 1 week.The actin bundles were stained with rhodamine-labeled phalloidin, and nu-clei were stained with DAPI.CD11b and CD18 antibodies, lentivirus with interfering vector plasmid of target gene Itgam (encoding CD11b) and empty virus (control virus) were used to treat osteoclasts for 1 week, and then immunofluorescence staining was performed.CD11b antibody, lentivirus and control virus were used to treat osteoclasts for 1 week, and total protein was taken for Western blot .RESULTS:Lower multinuclear positive rates in the groups treated with CD 11b anti-body were observed than that in the groups treated with CD 18 antibody and control group .Lower immunofluorescence inten-sity of Syk, CD11b and NFATc1 was found in CD11b antibody group than that in control group .Lower Syk, CD11b and NFATc1 immunofluorescence intensity was also observed in lentivirus group than that in control virus group .The results of Western blot analysis showed that the protein levels of CD 11b, Syk, NFATc1, c-Fos and p-ERK/ERK in CD11b antibody group were decreased as compared control group .Compared with control virus group , the protein levels of CD11b, Syk, NFATc1, c-Fos and p-ERK/ERK in lentivirus group were also decreased .CONCLUSION:CD11b subunit of Mac-1 pro-motes osteoclast differentiation by up-regulating c-Fos, ERK activity and NFATc1.

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