Effect Of Magnoliae Acupoint Application On Intestinal Flora In Patients With Hepatocellular Carcinoma And Its Promote Postoperative Gastrointestinal Function Recovery

Objective The aim of this study was to explore effect of magnoliae acupoint application on intestinal flora in patients with Hepatocellular Carcinoma(HCC)and its promote postoperative gastrointestinal function recovery.It provides a new idea for further clinical exploration of the mechanism of magnoliae acupoint application promote postoperative gastrointestinal function recovery in patients with HCC.Methods1.Study subjects and intervention methodsPatients admitted to the department of hepatobiliary surgery,the first affiliated hospital of Guangxi medical university from February 2019 to December 2019,underwent partial hepatectomy at selected time,and were pathologically diagnosed with HCC after liver resection were selected as the study subjects.This study is a optimum effectiveness test。Therefore,the total sample size of 99 cases was calculated by the optimum effectiveness test formula,and the HCC patients meeting the inclusion criteria were randomly divided into three groups: magnoliae acupoint application group,oral probiotics group and routine control group,with 33 cases in each group.On the basis of the routine control group,magnoliae acupoint application group was applied magnoliae acupoint application on the day 1 to day 4 after hepatic resection.The intervention frequency is once a day and each intervention time is 9:00 pm-9:00 am.In the oral probiotics group,on the basis of the routine control group,live combined bifidobacterium and lactobacillus tablets were taken on the day 1to day 4 after hepatic resection.Taking the medicine 3 times a day and the quantity of each time is 4 tablets.The routine control group practiced the routine nursing method of hepatobiliary surgery after hepatic resection.2.Specimen collection and detection method of main strainsBefore operation and the 5th days after hepatectomy,patients were asked to take stool samples.The total DNA of feces samples were extracted by a TIANamp Stool DNA Kit(Tiangen,DP328-02).Then the absolute quantitative method of real time fluorescent quantitative PCR was performed using the TB Green TM Premix Ex Taq TM Ⅱ(Tli RNase H Plus)(Ta Ka Ra,RR820A)in a real-time fluorescent quantitative PCR(Applied Biosystems7500).The amplified bacteria included bifidobacterium,the main beneficial bacteria,and enterobacteriaceae,the main opportunistic pathogen.The specificity of primer was determined by melting curve of the amplified product.When the melting curve of the amplified product is unimodal,the specificity of the primer is indicated.The standard curve was constructed by results of real-time fluorescence quantitative PCR amplification and DNA copy number of plasmid standard.Finally,the copy Numbers of bifidobacterium and enterobacterium in HCC patients were calculated by standard curves.3.Observe indicatorsChanges of the major strains before operation and after intervention: the main beneficial bacteria: bifidobacterium;the major opportunistic pathogens:enterobacteriaceae.Postoperative gastrointestinal function recovery index: first anal exhaust time,first anal defecation time,abdominal distension degree and abdominal distension score,nausea,vomiting and diarrhea.The first anal exhaust time and the first anal defecation time were divided into < mean and > mean according to the mean value.The extent of chang in prealbumin(PA)after operation.4.Statistical analysisSPSS 22.0 statistical software was used for statistical analysis.Measurement data such as age,Ishak score: inflammation score and fibrosis score,albumin were expressed as mean±standard deviation.Count data such as gender,smoking history,drinking history and ordinal data such as Barcelona clinical liver cancer stage,Chinese stage,postoperative abdominal distension were expressed as frequency(component proportion).Comparison of HCC patients among 3 groups: one-way analysis of variance was used for measurement data,chi-square test of row x list or Fisher exact probability method was used for counting data,and multiple samples rank sum test was used for ordinal data.Comparison between any two samples: LSD-t test was used for measurement data,Scheffe confidence interval method was used for counting data,and the result of confidence interval did not contain 0,indicating P<0.05.Nemenyi method was used for ordinal data.Comparison of HCC patients between two groups: the t-test for comparison of two samples was used for measurement data,the four tables chi-square test or Fisher exact probability test was used for count data,and the rank-sum test for comparison of two samples was used for ordinal data.Correlation analysis: Pearson correlation was used for measurement data and Spearman rank correlation was used for ordinal data.The changes in intestinal flora of HCC patients in each group before and after intervention and surgery were analyzed by paired t test.Propensity score matching was used to control confounding factors between groups,to equalize and compare between groups.Logistic regression was used to calculate the propensity score of HCC patients,and then 1:1 case matching was performed according to the nearest neighbor matching.The difference was statistically significant when P < 0.05.Results1.Specificity of primer and construction of standard curveThe real time fluorescence quantitative PCR amplification results showed that the melt curves of the amplification products all showed a single peak,indicating that the primers were specific,that is,only bifidobacterium was amplified from the primers of bifidobacterium,and only enterobacteriaceae was amplified from the primers of enterobacteriaceae.The standard curves of bifidobacterium and enterobacteriaceae constructed according to the results of PCR amplification and DNA copy number of plasmid standard both reached the quantitative standard.2.Changes of major strains of HCC patients before operation and after intervention in the 3 groupsThe abundance of bifidobacterium of HCC patients in the routine control group after intervention was significantly lower than before operation(P <0.05).However,the change of the abundance of bifidobacterium was no statistical difference after intervention compare with before operation in the magnoliae acupoint application group and oral probiotics group(P>0.05).The change of the abundance of enterobacteriaceae was no statistical difference after intervention compare with before operation in 3 group(P>0.05).It was shown that both magnoliae acupoint application and oral probiotics could promote the recovery of bifidobacterium effectively after HCC surgery,and the effects of the two methods in regulating intestinal flora were similar.3.Situation of postoperative gastrointestinal function recovery of HCC patients in 3 groupsA total of 90 eligible patients with HCC were eventually included in this study,30 cases in each group.Compared with routine control group,the first anal exhaust time was earlier,the proportion of exhaust time < mean was higher after operation in magnoliae acupoint application group and oral probiotic group,and the degree and score of abdominal distension on Day 3 were lower(P<0.05).There was no significant difference in gastrointestinal function recovery between magnoliae acupoint application group and oral probiotics group(P>0.05).4.Relationship between abdominal distension degree,abdominal distension score on the day 3 after surgery and postoperative first anal exhaust time in HCC patientsThe degree of abdominal distension(r=0.344,P<0.05),abdominal distension score(r=0.361,P<0.05)on the day 3 after surgery were positively correlated with the first postoperative anal exhaust time in HCC patients.5.Relationship between postoperative first anal exhaust time and major strains in HCC patientsThe first anal exhaust time were divided into group of < mean and >mean according to the mean value.Exhaust time > mean group showed that the abundance of bifidobacterium after liver resection is significantly lower than before liver resection(P<0.05).However,the abundance of bifidobacterium in the exhaust time < mean group before and after liver resection did not change significantly(P>0.05).The change in the abundance of enterobacteriaceae before and after the hepatic operation was no significant in this two groups(P>0.05).It suggested that early of postoperative exhaust was associated with postoperative recovery of bifidobacterium.6.Situation of postoperative PA change of HCC patients in 3 groupsThe postoperative PA reduction of HCC patients in magnoliae acupoint application group and oral probiotics group was lower than routine control group,but the difference was not statistically significant(P>0.05).Conclusions1.Magnoliae acupoint application can promote postoperative recovery of bifidobacterium effectively in HCC patients,play the role of beneficial bacteria in regulating intestinal flora.2.Magnoliae acupoint application can promote postoperative recovery of gastrointestinal function effectively in HCC patients,the pathway of action is related to the regulation of intestinal flora by beneficial bacteria.It provides a new idea for clinical further exploration of the mechanism of herbal acupoint application to promote postoperative gastrointestinal function recovery in patients.3.Magnoliae acupoint application is simple,convenient,inexpensive,and effective.And is a measure to give full play to the characteristics of external treatment of traditional Chinese medicine and carry forward the traditional Chinese medicine,which is worthy of further clinical exploration and application.