1.Study on molecular marker screening and evaluation of Rubi Fructus and its analogues
Mengdi ZHENG ; Lingling ZHANG ; Qi ZHANG ; Long GUO ; Xingjun WANG
International Journal of Traditional Chinese Medicine 2024;46(6):792-796
Objective:To standardize the origin of Rubi Fructus by using ITS2 and matK molecular markers to identify Rubi Fructus and its analogues. Methods:The ITS and matK sequences of Rubus chingii Hu, Rubus corchorifolius L.f., Rubus hirsutus Thunb., Rubus parvifolius L., Rubus buergeri Miq. and Rubus lambertianus Ser. were amplified and sequenced. The hidden Markov model was used to remove 5.8S and 28S from ITS sequences. A total of 25 ITS2 sequences were obtained. And a total of 22 matK sequences were obtained by proofreading by Glustal software. MEGA software was used for matK and ITS2 sequences analysis, intraspecific and interspecific genetic distances caculation, and neighbor joining (NJ) phylogenetic tree construction. ITS2 secondary structure was predicted by ITS2 database and aligned by 4Sale software. Profile neighbor-joining (PNJ) phylogenetic tree was constructed based on combined ITS2 sequence and its secondary structure by ProfDistS software. Results:An obvious barcoding gap between Rubi Fructus and its analogues was showed. The topological relationship between NJ tree and PNJ tree was consistent, and each taxon exhibits monophyly. The ITS2 secondary structure of Rubi Fructus was significant different from its analogues.Conclusions:It is recommended that both ITS2 and matK markers can serve as DNA barcodes for identifying Rubi Fructus and its analogues. The addition of ITS2 secondary structure information can enrich the identification results and provide theoretical support for resource research and variety selection of Rubi Fructus.
2.Perioperative efficacy of laparoscopic duodenal-preserving pancreatic head resection
Taoyuan YIN ; Xiaoxiang WANG ; Hang ZHANG ; Xingjun GUO ; Min WANG ; Renyi QIN
Chinese Journal of Surgery 2024;62(7):671-676
Objective:To compare the perioperative outcomes of laparoscopic duodenal-preserving pancreatic head resection(LDPPHR) with laparoscopic pancreaticoduodenectomy(LPD) in the treatment of borderline and benign diseases of the pancreatic head.Methods:This is a retrospective cohort study. Perioperative data from 87 patients with non-malignant pancreatic head diseases who underwent LDPPHR or LPD were retrospectively collected in the Department of Biliary-Pancreatic Surgery,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology from January 2020 to December 2022. There were 49 male and 38 female patients with an age ( M(IQR)) of 57.0(16.5) years (range: 20 to 75 years). Forty patients underwent LDPPHR and 47 patients underwent LPD. Quantitative data following a normal distribution were compared using Student′s t-test, while quantitative data not following a normal distribution were compared using the Mann-Whitney U test. Comparisons of categorical or ordinal variables were made using χ2 test or Fisher′s exact test. Logistic regression analysis was used to estimate the risk factors associated with the rate of complications. Results:There were no statistically significant differences between the LDPPHR group and the LPD group in terms of reoperation rate,total hospital stay duration,postoperative hospital stay duration,90-day mortality rate,30-day and 90-day readmission rates,and 2-year tumor recurrence rate (all P>0.05). The complication rate was higher in the LDPPHR group compared to the LPD group (80.0%(32/40) vs. 51.1%(24/47), χ2=7.89, P=0.005),but there was no difference in the rate of Clavien-Dindo classification of surgical complications ≥Ⅲ between the two groups (10.0%(4/40) vs. 12.8%(6/47), χ2<0.01, P=0.947). Additionally,the rate of delayed gastric emptying (DGE) was higher in the LDPPHR group compared to the LPD group ( χ2=10.79, P=0.001),but there was no statistically significant difference in the rate of B,C grade DGE between the two groups ( χ2=0.48, P=0.487). There were no statistically significant differences in the rates of postoperative pancreatic fistula,bile leakage,post-pancreatectomy hemorrhage,intra-abdominal infection,and pulmonary infection between the two groups (all P>0.05). The results of the univariate logistic regression analysis showed that LDPPHR (compared to LPD, OR=3.83, 95% CI: 1.46 to 10.04, Z=2.73, P=0.006) and preoperative biliary stent placement (compared to non-use of biliary stent, OR=5.30, 95% CI: 1.13 to 25.00, Z=2.11, P=0.035) were risk factors for the complication rate,but neither was an independent risk factor for complication rate (all P>0.05). Conclusion:The preliminary results suggest that LDPPHR can achieve perioperative safety and effectiveness comparable to LPD.
3.Perioperative efficacy of laparoscopic duodenal-preserving pancreatic head resection
Taoyuan YIN ; Xiaoxiang WANG ; Hang ZHANG ; Xingjun GUO ; Min WANG ; Renyi QIN
Chinese Journal of Surgery 2024;62(7):671-676
Objective:To compare the perioperative outcomes of laparoscopic duodenal-preserving pancreatic head resection(LDPPHR) with laparoscopic pancreaticoduodenectomy(LPD) in the treatment of borderline and benign diseases of the pancreatic head.Methods:This is a retrospective cohort study. Perioperative data from 87 patients with non-malignant pancreatic head diseases who underwent LDPPHR or LPD were retrospectively collected in the Department of Biliary-Pancreatic Surgery,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology from January 2020 to December 2022. There were 49 male and 38 female patients with an age ( M(IQR)) of 57.0(16.5) years (range: 20 to 75 years). Forty patients underwent LDPPHR and 47 patients underwent LPD. Quantitative data following a normal distribution were compared using Student′s t-test, while quantitative data not following a normal distribution were compared using the Mann-Whitney U test. Comparisons of categorical or ordinal variables were made using χ2 test or Fisher′s exact test. Logistic regression analysis was used to estimate the risk factors associated with the rate of complications. Results:There were no statistically significant differences between the LDPPHR group and the LPD group in terms of reoperation rate,total hospital stay duration,postoperative hospital stay duration,90-day mortality rate,30-day and 90-day readmission rates,and 2-year tumor recurrence rate (all P>0.05). The complication rate was higher in the LDPPHR group compared to the LPD group (80.0%(32/40) vs. 51.1%(24/47), χ2=7.89, P=0.005),but there was no difference in the rate of Clavien-Dindo classification of surgical complications ≥Ⅲ between the two groups (10.0%(4/40) vs. 12.8%(6/47), χ2<0.01, P=0.947). Additionally,the rate of delayed gastric emptying (DGE) was higher in the LDPPHR group compared to the LPD group ( χ2=10.79, P=0.001),but there was no statistically significant difference in the rate of B,C grade DGE between the two groups ( χ2=0.48, P=0.487). There were no statistically significant differences in the rates of postoperative pancreatic fistula,bile leakage,post-pancreatectomy hemorrhage,intra-abdominal infection,and pulmonary infection between the two groups (all P>0.05). The results of the univariate logistic regression analysis showed that LDPPHR (compared to LPD, OR=3.83, 95% CI: 1.46 to 10.04, Z=2.73, P=0.006) and preoperative biliary stent placement (compared to non-use of biliary stent, OR=5.30, 95% CI: 1.13 to 25.00, Z=2.11, P=0.035) were risk factors for the complication rate,but neither was an independent risk factor for complication rate (all P>0.05). Conclusion:The preliminary results suggest that LDPPHR can achieve perioperative safety and effectiveness comparable to LPD.
4.The importance of "Four Beams and Eight Pillars" matching theory
Yan ZHANG ; Shuilong MAO ; Zengjun GUO ; Han ZHANG ; Xinxin ZHANG ; Boxin JING ; Ping YAN ; Xingjun WANG
International Journal of Traditional Chinese Medicine 2022;44(8):846-848
This paper summarizes the current situation and problems of the research on the theory of "Taibai Seven Medicines" through literature summary and surveys. Although the "Four Beams and Eight Pillars" theory of compounding has a long history, the current research progress of "Taibai Seven Medicines" is mostly focused on plant resources. There lacks researches on the theory of compounding, or development of compounding and large varieties based on "Taibai Seven Medicines". We call for the inheritance and protection of folk experience, and hope that the "Four Beams and Eight Pillars" theory can guide the application and development of the seven Taibai medicines. Therefore, increasing the research on the theory and teaching in undergraduate courses formulary in Chinese medicine are necessary in our province.
5.Effect of miR-548j-5p expression on invasion of pancreatic cancer
Ruizhi HE ; Feng PENG ; Yechen FENG ; Xingjun GUO ; Xu LI ; Min WANG ; Rui TIAN ; Feng ZHU ; Renyi QIN
Chinese Journal of Hepatic Surgery(Electronic Edition) 2018;7(1):82-86
Objective To explore the expression of miR-548j-5p in pancreatic cancer and its effect on the invasion of pancreatic cancer. Methods Tissue samples of 16 patients with pancreatic cancer who received treatment in Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science &Technology between January 2016 and January 2017 were collected. There were 8 males and 8 females, aged from 41 to 62, including 7 cases with an age of ≤50 and 9 cases with an age of >50. The informed consents of all patients were obtained and the local ethical committee approval was received. The expression of miR-548j-5p in pancreatic cancer and adjacent tissues was detected by RT-PCR. PANC-1 cells were transfected by mimic and inhibitor of miR-548j-5p to construct PANC-1 cells with over-expression and low-expression of miR-548j-5p. And in control group, PANC-1 cells were transfected with empty plasmid Ctrl. The effect of miR-548j-5p on PANC-1 invasion was detected by Transwell assay, and its effect on PANC-1 metastasis was evaluated by wound scratch assay. Comparison of three groups was conducted by one-way analysis of variance and LSD-t test. Results The median relative expression of miR-548j-5p in pancreatic cancer was 0.35(0.03-4.47), significantly lower than 4.30 (0.04-65.55) in adjacent tissues (Z=-2.689, P<0.05).Transwell assay results showed that the average number of cells penetrating membrane was(58±34) in miR-548j-5p over-expression group, significantly lower than (231±61) in control group (LSD-t=-2.852, P<0.05);and it was(491±149)in miR-548j-5p low-expression group,significantly higher than that in control group (LSD-t=4.313, P<0.05). The wound scratch assay results showed that the wound scratch healing rate was (20±8)% in miR-548j-5p over-expression group, significantly lower than (38±6)% in control group (LSD-t=-3.759, P<0.05); and it was (53±6)% in miR-548j-5p low-expression group, significantly higher than that in control group (LSD-t=2.946, P<0.05). Conclusions Low expression of miR-548j-5p is observed in pancreatic cancer. Over expression of miR-548j-5p can decrease the invasive and metastatic ability of pancreatic cancer.
6.Clinical value of arterial first approach in laparoscopic pancreaticoduodenectomy
Chunyang MA ; Feng ZHU ; Min WANG ; Feng PENG ; Hang ZHANG ; Xingjun GUO ; Yechen FENG ; Hebin WANG ; Renyi QIN
Chinese Journal of Digestive Surgery 2017;16(8):832-838
Objective To investigate the clinical value of arterial first approach in laparoscopic pancreaticoduodenectomy (LPD).Methods The retrospective cohort study was conducted.The clinicopathological data of 181 patients with pancreatic head and periampullay tumors who underwent LPD in the Affiliated Tongji Hospital of Huazhong University of Science and Technology between October 2014 and December 2016 were collected.Among 181 patients,96 using arterial first approach and 85 using traditional approach were respectively allocated into the experimental group and the control group.Surgery was applied to patients in the same doctors' team,and there were the same extent of surgical resection,range of lymph node dissection and digestive tract reconstruction.Observation indicators:(1) intraoperative situation;(2) postoperative situation;(3) followup and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect the tumor-free survival up to February 2017.Measurement data with normal distribution were represented as x±s,and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range).Comparison of count data were analyzed using the chi-square test or Fisher exact probability.Results (1) Intraoperative situation:all the patients underwent successful LPD.Overall operation time and time of digestive tract reconstruction were respectively (268 ± 20) minutes,(33 ± 10) minutes in the experimental group and (285±25)minutes,(30± 17)minutes in the control group,with no statistically significant difference between 2 groups (t =8.529,2.741,P> 0.05).Time of tumor resection with superior mesenteric venous invasion were respectively (216± 13)minutes and (264±22)minutes in the experimental and control groups,with a statistically significant difference between the 2 groups (t=41.826,P<0.05).Time of tumor resection without superior mesenteric venous invasion were respectively (224± 14) minutes and (215±21) minutes in the experimental and control groups,with no statistically significant difference between the 2 groups (t =7.423,P> 0.05).Volumes of intraoperative blood loss and blood transfusion were respectively (99± 16)mL,(1.3±0.8)U in the experimental group and (131±27)mL,(2.8±1.2)U in the control group,with statistically significant differences between the 2 groups (t =3.670,0.562,P< 0.05).Five and 8 patients had intraoperative blood transfusion in the experimental and control groups,showing no statistically significant difference between the 2 groups (x2=1.195,P>0.05).(2) Postoperative situation:time of drainage tube removal and duration of hospital stay were respectively (5.8±2.4)days,(18.3±6.3) days in the experimental group and (6.3±3.6)days,(19.6±7.1) days in the control group,with no statistically significant difference between the 2 groups (t =0.498,1.305,P>0.05).Eleven patients in the experimental group had postoperative early complications,including 8with grade A pancreatic fistula (4 combined with diarrhea,2 combined with biliary fistula,1 combined with delayed gastric emptying and 1 with single pancreatic fistula),3 with grade B pancreatic fistula (2 combined with intra-abdominal hemorrhage and 1 combined with intra-abdominal infection).One patient with intra-abdominal hemorrhage in the experimental group died after treatment failure.Twelve patients in the control group had postoperative early complications,including 6 with grade A pancreatic fistula (2 combined with biliary fistula,2 combined with delayed gastric emptying,1 combined with diarrhea,1 combined with digestive tract hemorrhage),3 with grade B pancreatic fistula and intra-abdominal hemorrhage (2 combined with infection,including 1 death) and 3 with diarrhea.Other patients with complications were cured by symptomatic and supportive treatment.There was no statistically significant difference in overall complications between the 2 groups (x2 =0.287,P>0.05).Results of postoperative pathological examination showed that case with R0 resection was 93 and 76 in the experimental and control groups,with a statistically significant difference between the 2 groups (x2 =4.057,P<0.05).(3) Follow-up and survival situations:179 patients were followed up for 2-28 months,with a median time of 14 months.Postoperative 6-month tumor-free survival rate was 92.7% (89/96) and 88.2%(75/85) in the experimental and control groups,with no statistically significant difference between the 2 groups (x2=1.060,P>0.05).Conclusion Arterial first approach in LPD could significantly shorten the time of tumor resection of patients with superior mesenteric artery invading pancreatic head and periampullay region,significantly reduce the volumes of intraoperative blood loss and blood transfusion,and increase the rate of R0 resection.
7.Observation of the results in the treatment of Sequential hepatic arteria chemoembolization,percuta-neous radiofrequency ablation for hepatocellular carcinoma
Xiaobin ZHANG ; Xing WANG ; Yanhong ZHENG ; Xingjun GUO
Journal of Chinese Physician 2016;(z1):49-51
Objective To investigate the efficacy and safety of combination of transcatheter arterial chemoembolization (TACE)and percutaneous radiofrequency ablation (RFA)in treatment of hepatocellular carcinoma (PHC).Methods 70 cases of PHC were divided into combined group (TACE +RFA,n =37) and control group (only TACE,n =33).Patients were followed up for1 to 2 years,and the therapeutic effects and side effects were compared between the two groups.Results The patients with tumor completely necrosis and AFP level lower than >50% in combined group were significantly more than those in control group (P <0.05);the half of year,one year and two years survival rate in combined group were greatly higher than those of control group;no severe side effect was observed in two groups.Conclusions TACE+RFA is effective and safe in treatment of PHC of more than 5 cm in diameter.
8.Simulation of acoustic response of microvessel containing microbubble in ultrasound field based on finite element analysis and lumped parameter model
Chuanxiao NIU ; Shengwen GUO ; Lin QIU ; Yonghua LAO ; Xingjun JIANG
Chinese Journal of Tissue Engineering Research 2016;20(42):6330-6336
BACKGROUND:Exploration on nonlinear acoustic response of the contrast agent microbubble contained in microvessel under ultrasound excitation is of great significance to maximizing ultrasonic energy deposition, promoting the development of quantitative imaging algorithm, revealing the damage mechanism or evaluating the targeted therapy, and overcoming the limitations of the traditional methods that are mainly used in large-size vessels, and measuring microvessel elasticity.
OBJECTIVE:To build a microvessel containing an ultrasound microbubble, revealing the internal mechanism among ultrasound, microbubble, blood flow and microvessel.
METHODS:Based on the finite element analysis and the lumped parameter model, three-dimensional microvessel containing microbubble model was built and simulated on Comsol Multiphysics 4.4 platform.
RESULTS AND CONCLUSION:Microbubble exhibited slower radial motion compared with axial motion due to vascular wal limitation, but maximum displacement and stress were found near the microbubble center because of the oscil ation coupling of the microbubble with the vascular wal . Under the same ultrasound pressure, the excitation frequency increased, accompanied by decreased and stabilized microvessl constriction and dilation;under the same frequency, with the enhancement of ultrasound pressure, the local microbubble oscil ation lasted longer. With the increase of Young’s modulus of the microvessel wal , the frequency of microbubble oscil ation was reduced, while the amplitude increased. Al these findings indicate that the frequency of microbubble oscil ation increased with the reduction of microvessel size, while its amplitude decreased. The frequency of microbubble oscil ation increased with the enhancement of ultrasound excitation, while the amplitude decreased. On the contrary, ultrasound pressure affected the dynamic characteristics of microbubble and microvessel. In particular, it was the first to demonstrate that the elasticity of microvessel has approximate linear positive correlation with the amplitude of microbubble oscil ation, which reveals the relationship between microvessel elasticity and microbubble response so as to provide theoretical basis for indirect measurement of microvessel elasticity.
9.Effect of Administration Timing of Mannitol on Efficacy and Related Indexes of Patients with Moderate Cere-bral Hemorrhage
China Pharmacy 2016;27(21):2926-2927,2928
OBJECTIVE:To explore the effect of administration timing of mannitol on efficacy and related indexes of patients with moderate cerebral hemorrhage. METHODS:Data of 280 patients with moderate cerebral hemorrhage being treated with manni-tol were divided into super-early group(65 cases),early group(97 cases)and extension group(118 cases)was analyzed retrospec-tively according to administration timing. All patients received conservative medical treatment,ntrition brain cells and other symp-tomatic and supportive treatment. Based on it,super-early group received mannitol within 6 h of cerebral hemorrhage,early group received mannitol within 6-24 h,and extension group received mannitol more than 24 h. Clinical efficacy,incidence of hematoma enlargement,mortality,incidence of continued bleeding/rebleeding and incidence of adverse reactions in 3 groups were observed. RESULTS:The incidences of hematoma enlargement and mortality in early group and extension group were significantly lower than super-early group,the differences were statistically significant (P<0.05);but there were no significant differences between early group and extension group (P>0.05). The total effective rate in early group was significantly higher than super-early group and extension group,the incidence of continued bleeding/rebleeding was significantly lower than super-early group and extension group,the differences were statistically significant (P<0.05);but there no significant differences between super-early group and extension group (P>0.05). CONCLUSIONS:Based on conventional treatment,mannitol shows better efficacy for patients with moderate cerebral hemorrhage within 6-24 h of cerebral hemorrhage than those in super-early or extension. It can reduce continued bleeding/rebleeding incidence,and has equivalent safety.
10.Cloning and expression of a gene encoding shortened LfcinB(1-15)-Melittin(5-12) hybrid peptide in Escherichia coli BL21(DE3).
Chongpeng BI ; Xingjun FENG ; Anshan SHAN ; Jiayin GUO
Chinese Journal of Biotechnology 2009;25(7):975-981
In order to get new antibacterial peptide, we designed a hybrid peptide LfcinB(1-15)-Melittin(5-12), composed of 1-15 amino acid residues of bovine Lactoferricin and 5-12 amino acid residues of Melittin. According to the bias of codon utilization of Escherichia coli, We synthesized the gene encoding the hybrid peptide. We inserted the gene between the sites of Nco I and Sal I of pET-32a and obtained the recombinant expression vector for heterologous expression of LfcinB(1-15)-Melittin(5-12) in Escherichia coli. We used Escherichia coli BL21(DE3) as expression host for the recombinant plasmid. After induced by isopropyl-beta-D-thiogalactoside (IPTG) under the optimized conditions, we realized the fusion protein was successfully expressed. The fusion protein was expressed in soluble form and the level was more than 35% of the total proteins. With (His)6 x Tag, the fusion protein was easily purified by His x Bind Purification Kit. After purification, we obtained 35 mg of fusion protein from 1 L of culture medium. At last, we accomplished that the peptide LfcinB(1-15)-Melittin(5-12) was released from the fusion protein cleaved by enterokinase. The recombinant LfcinB(1-15)-Melittin(5-12) showed antimicrobial activity assayed by agar diffusion test. This is the first report on the heterologous expression of the hybrid antibacterial peptide LfcinB(1-15)-Melittin(5-12) in Escherichia coli and also provides basis for next cost-effective expression of other antimicrobial peptides in genetic engineering.
Animals
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Anti-Bacterial Agents
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biosynthesis
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Antimicrobial Cationic Peptides
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biosynthesis
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chemistry
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genetics
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Cattle
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Cloning, Molecular
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Escherichia coli
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genetics
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metabolism
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Genetic Vectors
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Lactoferrin
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biosynthesis
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genetics
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Melitten
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biosynthesis
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genetics
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Recombinant Fusion Proteins
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biosynthesis
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genetics

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