1.Anti-inflammatory effects and related mechanisms of naringenin in human periodontal ligament stem cells under lipopolysaccharide stimulation based on RNA sequencing
Junyu LI ; Xiaomei XU ; Xingyu LIU ; Ting ZENG ; Li ZHANG ; Qian ZHENG
West China Journal of Stomatology 2024;42(4):512-520
Objective RNA sequencing(RNA-seq)and bioinformatic analysis were combined and used to explore the anti-inflammatory effects and mechanisms of naringenin(Nar)in lipopolysaccharide(LPS)-stimulated human peri-odontal ligament stem cells(hPDLSCs).Methods Cell counting kit-8,quantitative real-time reverse transcription poly-merase chain reaction(qRT-PCR),and enzyme-linked immunosorbent assay(ELISA)were adopted to detect the effects of Nar on the proliferation and expression of inflammatory factors in LPS-stimulated hPDLSCs,screening for the opti-mal anti-inflammatory concentration of Nar.Differentially expressed genes(DEGs)were screened using|log2FC|≥1 and P≤0.05 as criteria.Volcano plot analysis,Kyoto En-cyclopedia of Genes and Genomes(KEGG)pathway en-richment analysis,the String database,and the MCODE module of Cytoscape were utilized to select core genes and enriched pathways.The effects on the nuclear factor κB(NF-κB)signaling pathway were verified using ELISA,qRT-PCR,and Western blot.Results Appropriate concentrations of Nar could alleviate the expression of inflammatory fac-tors and promote the proliferation of hPDLSCs stimulated by LPS.The best anti-inflammatory effect was achieved with 20 μmol/L Nar.RNA-seq showed significant enrichment of inflammation-related signaling pathways.The anti-inflamma-tory effect of Nar was mediated by inhibiting the NF-κB signaling pathway,similar to the effect of the NF-κB inhibitor BAY 11-7802.Conclusion Nar could exert its anti-inflammatory effects by inhibiting the NF-κB signaling pathway,making it a potential therapeutic option for the adjuvant treatment of periodontitis.
2.Effects of Acupuncture on Gut Microbiota and Central Iinflammation in Rat Model of Migraine based on"Gut-Brain"Theory
Mingsheng SUN ; Xingyu CHEN ; Mengdi ZHOU ; Yi LIU ; Yuqi ZHENG ; Siying HUANG ; Dingjun CAI ; Ling ZHAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(8):2726-2734
Objective Based on the theory of"gut-brain",this study explored the effect of acupuncture on the gut microbiota and central inflammation in migraine model rats,in order to explore the mechanism of acupuncture in the treatment of migraine from the perspective of"gut-brain".Methods The migraine rat model was established by subcutaneous injection of nitroglycerin.They were randomly divided into a model group and an acupuncture group,with 6 rats in each group,and a control group with 6 rats for conventional binding and fixation.Before modeling and on the 1st,5th,and 9th days after modeling,each group used electronic VonFrey to measure the plantar mechanical pain threshold of rats.After the experiment,Elisa was used to detect the expression levels of inflammatory factors IL-6 and TNF-α in the central trigeminal spinal nucleus of the rats in each group.Three-generation Pacbio full-length microbial diversity sequencing was used to perform 16S full-length rDNA sequencing on each group of fecal samples to detect the operational taxonomic unit(OTU)clustering and its abundance,Alpha diversity index,Beta diversity index,species among the samples in each group.differences in abundance.Results In migraine model rats,plantar mechanical pain threshold was significantly decreased(P<0.01),central IL-6 and TNF-α contents were significantly increased(P<0.01),and the structure and abundance of gut microbiota were abnormal.change(P<0.01).Continuous acupuncture treatment can significantly increase the plantar mechanical pain threshold in migraine rats(P<0.01),regulate the diversity of gut microbiota in migraine rats,increase Lactobacillus murine,and reduce the abundance of Lactobacillus enterobacteriaceae.degree(P<0.05),and decreased the levels of IL-6 and TNF-α in the central nervous system of migraine model rats(P<0.01).Conclusion Acupuncture can exert the"gut-brain"anti-inflammatory and analgesic effect of acupuncture in the treatment of migraine by regulating the gut microbiota structure and the expression of central IL-6 and TNF-α inflammatory factors in migraine model rats.
3.In vivo antioxidant activity of rabbiteye blueberry (Vaccinium ashei cv. 'Brightwell') anthocyanin extracts.
Jing WANG ; Xingyu ZHAO ; Jiawei ZHENG ; Daniela D HERRERA-BALANDRANO ; Xiaoxiao ZHANG ; Wuyang HUANG ; Zhongquan SUI
Journal of Zhejiang University. Science. B 2023;24(7):602-616
Blueberries are rich in phenolic compounds including anthocyanins which are closely related to biological health functions. The purpose of this study was to investigate the antioxidant activity of blueberry anthocyanins extracted from 'Brightwell' rabbiteye blueberries in mice. After one week of adaptation, C57BL/6J healthy male mice were divided into different groups that were administered with 100, 400, or 800 mg/kg blueberry anthocyanin extract (BAE), and sacrificed at different time points (0.1, 0.5, 1, 2, 4, 8, or 12 h). The plasma, eyeball, intestine, liver, and adipose tissues were collected to compare their antioxidant activity, including total antioxidant capacity (T-AOC), superoxide dismutase (SOD) activity and glutathione-peroxidase (GSH-PX/GPX) content, and the oxidative stress marker malondialdehyde (MDA) level. The results showed that blueberry anthocyanins had positive concentration-dependent antioxidant activity in vivo. The greater the concentration of BAE, the higher the T-AOC value, but the lower the MDA level. The enzyme activity of SOD, the content of GSH-PX, and messenger RNA (mRNA) levels of Cu,Zn-SOD, Mn-SOD, and GPX all confirmed that BAE played an antioxidant role after digestion in mice by improving their antioxidant defense. The in vivo antioxidant activity of BAE indicated that blueberry anthocyanins could be developed into functional foods or nutraceuticals with the aim of preventing or treating oxidative stress-related diseases.
Male
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Mice
;
Animals
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Antioxidants/pharmacology*
;
Blueberry Plants
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Anthocyanins/pharmacology*
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Mice, Inbred C57BL
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Superoxide Dismutase
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Plant Extracts/pharmacology*
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Superoxide Dismutase-1
4.Effect of self-made Yiqi Yangyin Decoction on immune function, tumor markers and toxic and side effects of chemotherapy in patients with advanced gastric cancer chemotherapy
Guangwei SHI ; Wendong JING ; Xingyu JIN ; Xiaoting ZHENG ; Shengbao WU
International Journal of Traditional Chinese Medicine 2022;44(2):154-158
Objective:To explore the effect of self-made Yiqi Yangyin Decoction on immune function, serum tumor markers and toxic and side effects of chemotherapy in patients with advanced gastric cancer chemotherapy.Methods:A total of 88 patients with advanced gastric cancer who met the inclusion criteria in the hospital between December 2018 and December 2020 were divided into two groups according to the random number table method, with 44 in each group. The control group was treated with chemotherapy, and the observation group was given self-made Yiqi Yangyin Decoction on the basis of the control group. The two groups were treated for 4 consecutive cycles with 3 weeks as 1 cycle. The Traditional Chinese Medicine (TCM) symptoms were scored before and after treatment, and the levels of IgG, IgA and IgM were detected by immunoturbidimetry. The levels of carcinoembryonic antigen (CEA) and cancer antigen 125 (CA125) were detected by ELISA. The gastrointestinal reactions, thrombocytopenia, anemia, leukopenia and neurotoxicity were recorded and evaluated during treatment, and the clinical efficacy was assessed.Results:The total effective rate was 81.8% (36/44) in observation group and 61.4% (27/44) in control group (χ 2=4.53, P<0.05). After treatment, the scores of stomachache and gastric distension, poor appetite, shortness of breath, and drooping spirit in observation group were significantly lower than those in the control group ( t=17.28, 11.91, 5.02, 5.65, all Ps<0.001) while the levels of serum IgG [(9.39±0.46)g/L vs. (8.54±0.23) g/L, t=10.96], IgA [(1.35±0.42) g/L vs. (1.07±0.15) g/L, t=6.90] and IgM [(0.92±0.09) g/L vs. (0.78±0.10) g/L, t=4.17] were significantly higher than those in the control group ( P<0.01). The levels of serum CEA [(9.07±1.01) μg/L vs. (14.89±2.13) μg/L, t=16.38] and CA125 [(24.87± 4.68) kU/L vs. (30.75±5.33) kU/L, t=5.50] were significantly lower than those in the control group ( P<0.01). During treatment, the toxic and side effects of gastrointestinal reactions, thrombocytopenia, anemia, leukopenia and neurotoxicity were significantly lower in the observation group than those in the control group ( Z=18.52, 2.54, 3.12, 3.84, 2.34, P<0.05). Conclusion:Self-made Yiqi Yangyin Decoction can improve the TCM symptoms, enhance the immunity, reduce the levels of serum tumor markers, and relieve the toxic and side effects of chemotherapy of patients undergoing chemotherapy for advanced gastric cancer.
5.Early numerical rating scale and Oswestry disability index in postmenopausal osteoporosis treated with denosumab
Qi WEI ; Miao ZHENG ; Chengwei WENG ; Keyu ZHU ; Xingyu JIN ; Weifeng LIU ; Dong ZHANG ; Qiaocheng ZHAI ; Peng ZHANG ; Youjia XU
Chinese Journal of Orthopaedics 2022;42(12):768-775
Objective:To compare the numerical rating scales (NRS) and Oswestry disability index (ODI) of denosumab in Chinese postmenopausal osteoporosis patients after 3 months, and analyze the early adverse reactions to provide reference for clinical diagnosis and treatment.Methods:Using a prospective study method, 260 patients with postmenopausal osteoporosis who were outpatients and inpatients in the Second Affiliated Hospital of Soochow University from September 2020 to October 2021 were selected, and general information, including age, height, weight, bone mineral density, history of fragility fractures, and use of anti-osteoporosis drugs. All subjects received denosumab 60 mg subcutaneously, and were given calcium and vitamin D at the same time. Pain was scored by NRS before treatment and 3 months after treatment, and functional improvement was assessed by ODI.Results:After 3 months of denosumab treatment in postmenopausal women with osteoporosis, among patients with different age groups, different degrees of osteoporosis, history of fragility fractures, and history of use of anti-osteoporosis drugs, NRS score and ODI score were lower than those before treatment, and the difference was statistically significant ( P<0.05). In addition, in patients with a history of fragility fractures (mainly vertebral fractures), the NRS scores and the ODI score decreased more significantly, and the difference was statistically significant ( P<0.05); the NRS score and ODI score decreased more significantly in patients with severe osteoporosis than in patients with osteoporosis, and the difference was statistically significant ( P<0.05); the BMD value of lumbar spine was negatively correlated with the reduction of NRS score before and after treatment ( P=0.042). In this study, 260 patients had musculoskeletal pain in 6 (2.3%), fatigue in 5 (1.9%), rash in 4 (1.5%), urinary tract infection in 2 (0.7%), and dizziness in 2 (0.7%), 2 case of fever (0.7%), 1 case of hypocalcemia (0.4%), a total of 22 cases of adverse reactions were reported, and the overall adverse reaction rate was 8.5%. Conclusion:Denosumab can improve pain symptoms and functional disability early in the clinical application of Chinese postmenopausal women with osteoporosis, and the incidence of adverse reactions is low. Especially for postmenopausal female osteoporosis patients with severe osteoporosis, low lumbar spine bone density, and a history of fragility fractures (mainly vertebral fractures), the application effect is more significant.
6.Efficacy of the biopsy strategy combining 6-core systematic and 3-core MRI-targeted biopsy in the detection of prostate cancer
Xiang TU ; Xingyu XIONG ; Chichen ZHANG ; Diming CAI ; Shengjiang BAI ; Weitao ZHENG ; Tianhai LIN ; Zhenhua LIU ; Lu YANG ; Qiang WEI
Chinese Journal of Urology 2022;43(12):914-919
Objective:To investigate the efficacy of the biopsy strategy combining 6-core systematic and 3-core MRI-targeted biopsy on prostate cancer (PCa) detection in biopsy-na?ve patients.Methods:The clinical data of 121 biopsy-na?ve patients who underwent transperineal prostate biopsy in West China Hospital of Sichuan University from July 2018 to January 2020 were retrospectively analyzed. The average age was (64.7±9.1) years old. Pre-biopsy prostate-specific antigen (PSA) was (12.4±7.5)ng/ml, f/t PSA was 0.13±0.05. Prostate volume was (43.1±26.1) ml and PASD was (0.35±0.27) ng/ml 2. The prostate-imaging and data system (PI-RADS) score of MRI before biopsy was reported to be 3 for 29 patients (24.0%), 4 for 54 patients (44.6%) and 5 for 38 patients (31.8%). All 121 patients underwent 12-core systematic biopsy combined with a 3-core or 5-core MRI-targeted biopsy, of which 61 patients underwent 3-core targeted biopsy and 60 underwent 5-core targeted biopsy. There was no significant difference in the pre-biopsy clinical data between the two groups ( P>0.05). A 6-core systematic biopsy was redefined as the results of 6 cores among the 12-core systematic biopsy. We compared the detection rates among the single 12-core systematic biopsy, 6-core systematic biopsy, MRI-targeted biopsy (3-core or 5-core), and different systematic biopsy combing with targeted biopsy for any PCa and clinically significant PCa, and we also analyzed the cumulative cancer detection rates for MRI-targeted biopsy of different cores. Results:Of the 121 patients in this study, the biopsy results were negative for 43 patients (35.5%) and positive for 78 (64.5%). The detection rate of clinically significant PCa was 55.4% (67/121). The detection rate of the 6-core systematic biopsy combined with MRI-targeted biopsy was 62.0% (75/121) for PCa and 55.4% (67/121) for clinically significant PCa, which was of no difference compared with that for the 12-core systematic biopsy combined with MRI-targeted biopsy ( P>0.05), but the 6-core systematic biopsy combined with MRI-targeted biopsy avoided the overdiagnosis of 3 patients with Gleason score 3+ 3. The detection rate of PCa for MRI-targeted biopsy was 57.9% (70/121), including 42.1% (51/121) for the first core, 55.4% (67/121) for the first two cores, and 57.9% (70/121) for the first three cores. Compared with the single-core targeted biopsy for suspicious lesions, the first 2-core targeted biopsy ( OR=1.7, 95% CI 1.0-2.8) and 3-core targeted biopsy ( OR=1.9, 95% CI 1.1-3.1) can significantly increase the detection rate of PCa, while the fourth or fifth core of targeted biopsy can not increase the detection rate additionally (60%, 36/60). Conclusion:For patients with suspected PCa, the prostate biopsy strategy combing 6-core systematic and 3-core MRI-targeted biopsy performs no inferior than the current 12-core systematic biopsy combined with MRI-targeted biopsy.
7.Efficacy and safety of domestic RECO flow restoration device in acute intracranial large-vessel occlusion
Jie CAO ; Hang LIN ; Min LIN ; Kaifu KE ; Yunfeng ZHANG ; Yong ZHANG ; Weihong ZHENG ; Xingyu CHEN ; Wei WANG ; Meng ZHANG ; Jinggang XUAN ; Ya PENG
Chinese Journal of Neuromedicine 2020;19(5):462-469
Objective:To explore the efficacy and safety of domestic RECO flow restoration device in acute intracranial large-vessel occlusion (LVO).Methods:This study was a multicenter, prospective, randomized, open, controlled trial; 136 patients with acute intracranial LVO at 7 Chinese stroke centers from February 2014 to August 2016 were randomly assigned into an experimental group (thrombectomy by RECO device, n=67) and a control group (thrombectomy by Solitaire device, n=69). The efficacy and safety of patients from the two groups were compared and analyzed. The primary efficacy end point was set as achievement of good recanalization (modified thrombolysis in cerebral infarction [mTICI] grading≥2); the secondary efficacy end points included good prognosis (modified Rankin scale scores≤2 90 d after thrombectomy), time from puncture to achieving good recanalization/time from puncture to final angiogram on condition that good recanalization was not gained, or mortality within 90 d of thrombectomy. The safety end points included any device-related serious adverse events, symptomatic intracerebral hemorrhage or serious adverse events within 24 of thrombectomy. Results:There was no statistically significant difference between the experimental group and the control group in successful rate of good recanalization (91.0% vs. 86.9%), good prognosis rate (62.7% vs. 46.4%), time from puncture to achieving good recanalization/time from puncture to final angiogram on condition that good recanalization was not gained([85.4±47.0] min vs. [89.9±53.3] min), and mortality within 90 d of thrombectomy (13.4% vs. 23.2%, P>0.05). There were no device-related serious adverse events in all patients. No significant differences were found in the incidences of symptomatic intracranial hemorrhage (1.5% vs. 7.4%) or serious adverse events (death [1.5% vs. 1.4%] and brain hernia [4.5% vs. 0.0%]) between the two groups ( P>0.05). Conclusion:The domestic RECO flow restoration device is an effective and safe mechanical thrombectomy stent retriver for acute intracranial LVO.
8. Feasibility and safety of the medial approach "four-step method" in the laparoscopic mobilization of splenic flexure
Zejian LYU ; Wulin WU ; Zhenbin LIN ; Weijun LIANG ; Junjiang WANG ; Jiabin ZHENG ; Xingyu FENG ; Guanfu CAI ; Deqing WU ; Yong LI
Chinese Journal of Gastrointestinal Surgery 2019;22(7):668-672
Objective:
To investigate the feasibility and safety of the medial approach "four-step method" in the laparoscopic mobilization of splenic flexure.
Methods:
A retrospective cohort study was performed. Clinical data of 157 colorectal cancer patients undergoing the medial approach "four-step method" in the laparoscopic mobilization of splenic flexure at Gastrointestinal Surgical Department of Guangdong Provincial People′s Hospital from July 2015 to June 2018 were retrospectively analyzed. Of 157 cases, 17 were transverse colon cancer, 94 were descending colon cancer, 25 were sigmoid cancer and 21 were rectal cancer; 89 were male and 68 were female; mean age was (61.8±10.3) years and mean body mass index was (23.2±3.7) kg/m2. The medial approach "four-step method" in the laparoscopic mobilization of splenic flexure was performed as follows: (1) The root vessels were treated with the "provocation" technique to expand the Toldt′s gap. This expansion was extended from the lateral side to the peritoneum reflex of left colonic sulcus, from the caudal side to the posterior rectal space, and from the cephalad side to the lower edge of pancreas. (2) The left colonic sulcus was mobilized, converging with the posterior Toldt′s gap. Mobilization was carried out from cephalad side to descending colon flexure, freeing and cutting phrenicocolic ligament and splenocolic ligament, and from caudal side to peritoneal reflex. (3) Gastrocolic ligament was moblized. Whether to enter the great curvature of stomach omentum arch when the gastrocolic ligament was cut, that was, whether to clean the fourth group of lymph nodes, should be according to the tumor site and whether serosal layer was invaded. (4) Transverse mesocolon was moblized and transected at the lower edge of the pancreatic surface, merging with the posterior Toldt′s gap, and from lateral side to lower edge of the pancreatic body, merging with the lateral left paracolonic sulcus. Safety and short-term clinical efficacy of this surgical procedure was summarized.
Results:
All the patients completed this procedure. During operation, 3 cases were complicated with organ injury, including 1 case of colon injury, 1 case of spleen injury and 1 case of pancreas injury. No operative death and conversion to open surgery was found. The average operation time was (147.5±35.1) minutes, the average intra-operative blood loss was (40.8±32.7) ml and the average number of harvested lymph node was (16.1±5.8), including (4.0±2.3) of positive lymph nodes. The first exhaust time after surgery was (41.3±20.6) hours, the fluid intake time was (1.5±1.3) days, the postoperative hospital stay was (5.2±2.3) days. Eight (5.1%) cases developed postoperative complications, and all were improved and discharged after conservative treatments. According to the TNM classification system, postoperative pathology revealed that 31 patients were stage I, 51 were stage II, 53 were stage III, 22 were stage IV.
Conclusion
The medial approach "four-step method" is safe and feasible, which can effectively decrease the operation difficulty of the laparoscopic mobilization of the splenic flexure.
9.Feasibility and safety of the medial approach "four?step method" in the laparoscopic mobilization of splenic flexure
Zejian LYU ; Wulin WU ; Zhenbin LIN ; Weijun LIANG ; Junjiang WANG ; Jiabin ZHENG ; Xingyu FENG ; Guanfu CAI ; Deqing WU ; Yong LI
Chinese Journal of Gastrointestinal Surgery 2019;22(7):668-672
Objective To investigate the feasibility and safety of the medial approach "four?step method" in the laparoscopic mobilization of splenic flexure. Methods A retrospective cohort study was performed. Clinical data of 157 colorectal cancer patients undergoing the medial approach "four?step method" in the laparoscopic mobilization of splenic flexure at Gastrointestinal Surgical Department of Guangdong Provincial People′s Hospital from July 2015 to June 2018 were retrospectively analyzed. Of 157 cases, 17 were transverse colon cancer, 94 were descending colon cancer, 25 were sigmoid cancer and 21 were rectal cancer; 89 were male and 68 were female; mean age was (61.8±10.3) years and mean body mass index was (23.2±3.7) kg/m2. The medial approach "four?step method" in the laparoscopic mobilization of splenic flexure was performed as follows: (1) The root vessels were treated with the "provocation"technique to expand the Toldt′s gap. This expansion was extended from the lateral side to the peritoneum reflex of left colonic sulcus, from the caudal side to the posterior rectal space, and from the cephalad side to the lower edge of pancreas. (2) The left colonic sulcus was mobilized, converging with the posterior Toldt′s gap. Mobilization was carried out from cephalad side to descending colon flexure, freeing and cutting phrenicocolic ligament and splenocolic ligament, and from caudal side to peritoneal reflex. (3) Gastrocolic ligament was moblized. Whether to enter the great curvature of stomach omentum arch when the gastrocolic ligament was cut, that was, whether to clean the fourth group of lymph nodes, should be according to the tumor site and whether serosal layer was invaded. (4) Transverse mesocolon was moblized and transected at the lower edge of the pancreatic surface, merging with the posterior Toldt′s gap, and from lateral side to lower edge of the pancreatic body, merging with the lateral left paracolonic sulcus. Safety and short?term clinical efficacy of this surgical procedure was summarized. Results All the patients completed this procedure. During operation, 3 cases were complicated with organ injury, including 1 case of colon injury, 1 case of spleen injury and 1 case of pancreas injury. No operative death and conversion to open surgery was found. The average operation time was (147.5±35.1) minutes, the average intra?operative blood loss was (40.8±32.7) ml and the average number of harvested lymph node was (16.1±5.8), including (4.0±2.3) of positive lymph nodes. The first exhaust time after surgery was (41.3±20.6) hours, the fluid intake time was (1.5 ± 1.3) days, the postoperative hospital stay was (5.2 ± 2.3) days. Eight (5.1%) cases developed postoperative complications, and all were improved and discharged after conservative treatments. According to the TNM classification system, postoperative pathology revealed that 31 patients were stage I, 51 were stage II, 53 were stage III, 22 were stage IV. Conclusion The medial approach "four?step method" is safe and feasible, which can effectively decrease the operation difficulty of the laparoscopic mobilization of the splenic flexure.
10.Feasibility and safety of the medial approach "four?step method" in the laparoscopic mobilization of splenic flexure
Zejian LYU ; Wulin WU ; Zhenbin LIN ; Weijun LIANG ; Junjiang WANG ; Jiabin ZHENG ; Xingyu FENG ; Guanfu CAI ; Deqing WU ; Yong LI
Chinese Journal of Gastrointestinal Surgery 2019;22(7):668-672
Objective To investigate the feasibility and safety of the medial approach "four?step method" in the laparoscopic mobilization of splenic flexure. Methods A retrospective cohort study was performed. Clinical data of 157 colorectal cancer patients undergoing the medial approach "four?step method" in the laparoscopic mobilization of splenic flexure at Gastrointestinal Surgical Department of Guangdong Provincial People′s Hospital from July 2015 to June 2018 were retrospectively analyzed. Of 157 cases, 17 were transverse colon cancer, 94 were descending colon cancer, 25 were sigmoid cancer and 21 were rectal cancer; 89 were male and 68 were female; mean age was (61.8±10.3) years and mean body mass index was (23.2±3.7) kg/m2. The medial approach "four?step method" in the laparoscopic mobilization of splenic flexure was performed as follows: (1) The root vessels were treated with the "provocation"technique to expand the Toldt′s gap. This expansion was extended from the lateral side to the peritoneum reflex of left colonic sulcus, from the caudal side to the posterior rectal space, and from the cephalad side to the lower edge of pancreas. (2) The left colonic sulcus was mobilized, converging with the posterior Toldt′s gap. Mobilization was carried out from cephalad side to descending colon flexure, freeing and cutting phrenicocolic ligament and splenocolic ligament, and from caudal side to peritoneal reflex. (3) Gastrocolic ligament was moblized. Whether to enter the great curvature of stomach omentum arch when the gastrocolic ligament was cut, that was, whether to clean the fourth group of lymph nodes, should be according to the tumor site and whether serosal layer was invaded. (4) Transverse mesocolon was moblized and transected at the lower edge of the pancreatic surface, merging with the posterior Toldt′s gap, and from lateral side to lower edge of the pancreatic body, merging with the lateral left paracolonic sulcus. Safety and short?term clinical efficacy of this surgical procedure was summarized. Results All the patients completed this procedure. During operation, 3 cases were complicated with organ injury, including 1 case of colon injury, 1 case of spleen injury and 1 case of pancreas injury. No operative death and conversion to open surgery was found. The average operation time was (147.5±35.1) minutes, the average intra?operative blood loss was (40.8±32.7) ml and the average number of harvested lymph node was (16.1±5.8), including (4.0±2.3) of positive lymph nodes. The first exhaust time after surgery was (41.3±20.6) hours, the fluid intake time was (1.5 ± 1.3) days, the postoperative hospital stay was (5.2 ± 2.3) days. Eight (5.1%) cases developed postoperative complications, and all were improved and discharged after conservative treatments. According to the TNM classification system, postoperative pathology revealed that 31 patients were stage I, 51 were stage II, 53 were stage III, 22 were stage IV. Conclusion The medial approach "four?step method" is safe and feasible, which can effectively decrease the operation difficulty of the laparoscopic mobilization of the splenic flexure.

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