Objective:To investigate the efficacy of our self-made distractor in assistance of percutaneous minimally invasive treatment of Sanders Ⅱ and Ⅲ calcaneal fractures.Method:A retrospective study was conducted to analyze the clinical data of 51 patients with calcaneal fracture who had been treated from March 2020 to August 2022 at Department of Orthopedic Trauma, Changsha Central Hospital Affiliated to University of South China. There were 42 males and 9 females with an age of (44.6±10.2) years. All the 34 Sanders type Ⅱ fractures and 17 Sanders type Ⅲ fractures were isolated unilateral ones which were treated by percutaneous reduction and fixation with percutaneous screws assissted by our self-made calcaneal distractor. All patients were evaluated both clinically and radiologically. The interval from injury to surgery, surgery duration, hospitalization period, fracture union, complications, reduction of the posterior inferior articular surface, the American Orthopaedic Foot & Ankle Society (AOFAS) ankle-hindfoot score and visual analog scale (VAS) pain score were recorded. The length, width, height, B?hler angle, and Gissane angle of the calcaneus were measured and compared between pre-surgery and the last follow-up.Results:All patients in this cohort were followed up after uneventfull surgery for 21 (18, 24) months. The interval from injury to surgery was 0 (0, 1) d, the surgery duration 38 (35, 40) min, and the hospitalization period 4 (3, 4) d. All the surgical incisions healed smoothly without any wound-related complications. All fractures got united by the 12-week postoperative assessment with normal alignment of the posterior foot and no varus or valgus. One patient developed symptoms of sural nerve injury after surgery, and one patient experienced a screw rupture after surgery. Evaluation of the posterior subtalar articular surfaces revealed an anatomic reduction rate of 66.7% (34/51), a near-anatomical reduction rate of 33.3% (17/51), and no cases of approximate reduction or failure in articular surface reduction. The last follow-up found no loss of reduction. At the last follow-up, the AOFAS ankle-hindfoot score was 91.3 (87.0, 98.0) points, giving 32 excellent, 17 good, and 2 fair cases, resulting in an excellent and good rate of 96.1%, and the VAS pain score 0 (0, 1) point. At the last follow-up, the width [34.0 (32.6, 34.9) mm], height [49.6 (47.1, 50.4) mm], B?hler angle [35.8 (34.0, 37.8)°], and Gissane angle [129.0 (124.2, 135.6)°] of the calcaneus were all significantly improved compared with the preoperative values [(41.0±2.1) mm, 39.3 (37.9, 40.2) mm, -5.7 (-20.4, 4.6)°, 94.5 (80.4, 104.0)°] ( P < 0.05). There was no statistically significant difference in the length of the calcaneus between pre-surgery and post-surgery ( P > 0.05). Conclusion:In the percutaneous minimally invasive treatment of Sanders Ⅱ and Ⅲ calcaneal fractures, use of our self-made calcaneal distractor has demonstrated favorable clinical outcomes, making the surgery minimally invasive and efficient.

Blood deficiency syndrome (BDS) refers to a pathological state with blood dysfunction and organ dystrophy in traditional Chinese medicine. Danggui Wuji granules (DWG) was developed from a formula containing Angelicae Sinensis Radix and Musculus et Os Galli Domestici. Herein, we investigated the mechanism of DWG in treating BDS by modulating gut microbiota. We found that DWG protected mice from BDS by elevating the levels of red blood cell count, hemoglobin, and hematocrit in peripheral blood and increasing the erythrocyte membrane Na⁺-K⁺-ATPase activity. Danggui Wuji granules changed the composition and metabolites of colonic flora. Notably, Lactobacillus, Muribaculaceae, and Alistipes were the main genera showing changes after DWG treatment. Our findings revealed that DWG presented a positive therapeutic effect on BDS in mice by regulating the gut microbiota and metabolites. The protective mechanism of DWG was associated with pathways such as metabolic pathways, biosynthesis of secondary metabolites, ABC transporters, ribosome, thyroid hormone synthesis, lysine degradation, galactose metabolism, tyrosine metabolism, etc.

ObjectiveTo clarify the therapeutic effect of Huashi Baidu prescription on pneumonia in mice caused by influenza A （H1N1） virus and explore its mechanism based on the transcriptome. MethodA mouse influenza viral pneumonia model was built by intranasal infection with influenza A virus， and mice were continuously administered the drug for five days， so as to investigate the general condition， lung index， viral load， pathological morphology of lung tissue， survival time， and prolongation rate of survival time of mice and clarify the therapeutic effect of Huashi Baidu prescription on influenza viral pneumonia. Transcriptome technology was used to detect the differentially expressed genes in the lung tissue of mice in the model group and the normal group， as well as the Huashi Baidu prescription group and the model group， and the potential core target of the Huashi Baidu prescription for the treatment of influenza viral pneumonia was screened. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to verify the effect of Huashi Baidu prescription on the mRNA expression level of core target genes. ResultCompared with the normal group， the lung index and viral load in the lung tissue of the model group were significantly increased （P<0.05， P<0.01）. Compared with the model group， the high-dose group of Huashi Baidu prescription significantly prolonged the survival time of mice infected with influenza A virus （P<0.05） and significantly reduced the lung index value of mice （P<0.05） and the viral load of lung tissue. The high-dose， medium-dose， and low-dose groups of Huashi Baidu prescription could significantly reduce lung tissue inflammation， blood stasis， swelling， and other pathological changes in mice （P<0.05， P<0.01）. Transcriptome analysis of lung tissue showed that core genes were mainly enriched in the nuclear transcription factor-κB （NF-κB） signaling pathway， interleukin-17 （IL-17） signaling pathway， cytokine-cytokine receptor interaction， and other pathways after the intervention of Huashi Baidu prescription. TRAF6， NFKBIA， CCL2， CCL7， and CXCL2 were the top five node genes with combined score values. Real-time PCR validation showed that Huashi Baidu prescription significantly downregulated the mRNA expression of key genes TRAF6 and NFKBIA in the NF-κB signaling pathway， as well as chemokines CCL2， CCL7， and CXCL2 （P<0.05， P<0.01）. ConclusionHuashi Baidu prescription has a therapeutic effect on influenza viral pneumonia， possibly by inhibiting the expression of key nodes TRAF6 and NFKBIA in the NF-κB signaling pathway and that of chemokines CCL2， CCL7， and CXCL2， reducing the recruitment of inflammatory cells and viral load， and exerting anti-influenza viral pneumonia effects.

Objective To study the clinical use of microwave scalpel in hepatectomy for hepatocellular carcinoma without hepatic vascular inflow occlusion.Methods A retrospective study was conducted on the clinical data of 126 consecutive patients with hepatocellular carcinoma who were treated at the Department of Hepatobiliary Surgery of the Third Affiliated Hospital of Sun Yat-sen University from December 2015 to August 2018.The patients included 111 males and 15 females,with ages which ranged from 25 to 75 years.These patients were divided into two groups by the different surgical resection methods:the microwave scalpel hepatectomy (MSH) group which employed microwave scalpels to perform hepatectomy (n =42),and the ultrasound scalpel hepatectomy (USH) group which employed ultrasound scalpels (n =84).The perioperative conditions which included baseline data,amount of intraoperative bleeding,liver function after operation,and morbidity of postoperative complications were compared.Results There were no significant differences in the preoperative indexes which included tumor diameter,number of tumors,levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) between the 2 groups (all P ＞ 0.05).In the MSH group,the amount of intraoperative bleeding was 100.0 (100.0,200.0) ml,which was significantly lower than the 300.0 (100.0,400.0) ml in the USH group (P ＜ 0.05).Compared with the USH group,the levels of AST and ALT on postoperative day 1,3,7 after operation in the MSH group were significantly lower (all P ＜ 0.05),and the corresponding albumin levels were significantly higher than the USH group (P ＜0.05).The incidence of complications was 4.8％ (2/42) in the MSH group which was significantly lower than that of 20.2％ (17/84) in the USH group (P ＜ 0.05).Conclusion Microwave scalpel significantly reduced intraoperative bleeding and postoperative complications,and led to less liver functional injury.

Objective To analyze the effects of different therapies on patient survival,to explore the related prognostic factors in elderly patients with advanced gastric cancer,and to provide recommendations for the treatment of such patients.Methods Retrospective analysis was conducted on 146 elderly patients with advanced gastric cancer hospitalized from January 2009 to October 2013 in Yixing People's Hospital of Jiangsu Province.Detailed clinical data were recorded,and patients were followed up during the total survival time.Univariate analysis with the Log rank test and multivariate analysis with the COX proportional hazard model were utilized to examine the related prognostic factors in elderly patients with advanced gastric cancer.Results The 1-,2,3-year survival rates of t46 elderly patients with advanced gastric cancer were 33.6 ％,11.0 ％,and 2.1 ％,respectively,and the median survival time was 10.3 months.The Log-rank test showed that Karnofsky (KPS) score,differentiation degree,number of metastatic sites,malignant serous effusion,chemotherapy,and traditional Chinese medicine (TCM) were associated with the prognosis of elderly patients with advanced GC (all P＜0.05).Multivariate analysis by the COX proportional hazard model showed that KPS score (HR=1.575,95％ CI:1.094 2.267,P=0.015),differentiation degree (HR=0.499,95％CI:0.340-0.732,P＜0.001),malignant serous effusion (HR=0.516,95％ CI:0.356-0.748,P＜ 0.001),chemotherapy (HR=1.669,95％ CI:1.185 2.351,P=0.003),and TCM (HR=1.793,95％ CI:1.237-2.600,P=0.002) were independent factors related to the prognosis of elderly patients with advanced GC.Conclusions The prognosis of elderly patients with advanced gastric cancer is poor,especially for patients with a low KPS score,a poor differentiation degree,or malignant serous effusion.Chemotherapy and TCM can improve the prognosis.

Objective To investigate the influence of high + Gzenvironment on receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) messenger RNA (mRNA ) expression in the peri-implant bone tissue by an animal model. Methods Thirty New Zealand white rabbits were randomly and averagely divided into 3 control groups and 3 experimental groups corresponding to the investigation for 3 weeks ,5 weeks and 12 weeks respectively.Rabbits′ mandible incisors of both sides were extracted and im planted with 1 implant in each socket immediately.After 1 week of rest ,the experimental rabbits were exposed to high + Gzenvironment 3 times a week (Monday ,Wednesday ,Friday) ,The exposure scheme choose the + Gzprofile with 4～9 G for 10～45 s with 1 G/s onset rate and with the intervals of 1 min.The rabbits of control groups were not exposed to + Gzenvironment and normally fed.One control group and one experimental group rabbits were respectively sacrificed at the 3rd week (+Gzexposure for 2 weeks) , 5th week (+Gzexposure for 4 weeks) and 12th week (+Gzexposure for 4 weeks and normal feeding for 7 weeks) after the implanting surgery.The specimens were obtained to find the mRNA expression of RANKL and OPG by real-time polymerase chain reaction (RT-PCR) examination. Results The mRNA expression of RA NKL and OPG were the highest 3 weeks after the surgery in both groups , and then decreased gradually.The RANKL mRNA expression and the RANKL/OPG ratio of the experimental group were significantly higher than those of the control group 3 weeks after the surgery (P＜0.05).Five weeks after the surgery ,the OPG mRNA expression of the experimental group was significantlylowerthanthatofthecontrolgroup(P＜0.05),buttheRANKLmRNAexpressionand RANKL/OPG ratio were still higher than that of control group (P＜0.05).Twelve weeks after the surgery ,the differences on RANKL and OPG mRNA expressions was not obvious between two groups. Conclusions High +Gzexposure could increase the expression of RA N K L ,raise RA N K L/OPG ratio and promote bone resorption ,but may be not conducive to implant bone bonding.

Objective To investigate the influence of high + Gzenvironment on receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) messenger RNA (mRNA ) expression in the peri-implant bone tissue by an animal model. Methods Thirty New Zealand white rabbits were randomly and averagely divided into 3 control groups and 3 experimental groups corresponding to the investigation for 3 weeks ,5 weeks and 12 weeks respectively.Rabbits′ mandible incisors of both sides were extracted and im planted with 1 implant in each socket immediately.After 1 week of rest ,the experimental rabbits were exposed to high + Gzenvironment 3 times a week (Monday ,Wednesday ,Friday) ,The exposure scheme choose the + Gzprofile with 4～9 G for 10～45 s with 1 G/s onset rate and with the intervals of 1 min.The rabbits of control groups were not exposed to + Gzenvironment and normally fed.One control group and one experimental group rabbits were respectively sacrificed at the 3rd week (+Gzexposure for 2 weeks) , 5th week (+Gzexposure for 4 weeks) and 12th week (+Gzexposure for 4 weeks and normal feeding for 7 weeks) after the implanting surgery.The specimens were obtained to find the mRNA expression of RANKL and OPG by real-time polymerase chain reaction (RT-PCR) examination. Results The mRNA expression of RA NKL and OPG were the highest 3 weeks after the surgery in both groups , and then decreased gradually.The RANKL mRNA expression and the RANKL/OPG ratio of the experimental group were significantly higher than those of the control group 3 weeks after the surgery (P＜0.05).Five weeks after the surgery ,the OPG mRNA expression of the experimental group was significantlylowerthanthatofthecontrolgroup(P＜0.05),buttheRANKLmRNAexpressionand RANKL/OPG ratio were still higher than that of control group (P＜0.05).Twelve weeks after the surgery ,the differences on RANKL and OPG mRNA expressions was not obvious between two groups. Conclusions High +Gzexposure could increase the expression of RA N K L ,raise RA N K L/OPG ratio and promote bone resorption ,but may be not conducive to implant bone bonding.

Objective To explore the impact of telomerase regulation factor PinX1 to the proliferation and invasion ability of hepatoma cells. Methods Hepatoma cells PinX1-7721 (experimental group) with stable expression of PinX1 as well as control cell VECTOR-7721 (control group) were constructed. The expression of PinX1 mRNA was detected by RT-PCR. The proliferation ability and clonality of hepatoma cells were detected by CCK-8 method and plate clonality assay, and the invasion ability of hepatoma cells by Transwell assay. Comparison of the experiment data was conducted by t test. Results Expression level of PinX1 mRNA in experiment group was (13.9±2.0)×10-3, which was significantly higher than (1.1±0.2)×10-3in control group (t=10.98, P<0.05). A450of the cells on 1-7 d in experiment group was respectively 0.260±0.004, 0.340±0.008, 0.450±0.040, 0.500±0.020, 0.730±0.030, 1.350±0.040 and 1.640±0.050, which were significantly lower than 0.280±0.009, 0.410±0.007, 0.680±0.044, 0.730±0.029, 0.850±0.070, 1.700±0.020 and 2.080±0.280 in control group (t=-5.82, -12.99, -6.36, -5.96, -28.42,-18.98, -5.08; P<0.05). The plate clonality assay results showed that the clone formation quantity of cells in experiment group was 143±32, which was significantly lower than 305±25 in control group (t=-6.91, P<0.05).Transwell assay results showed that the quantity of trans-membrane cell in experiment group was 230±16, which was significantly lower than 650±30 in control group (t=-21.40, P<0.05). Conclusion PinX1 could inhibit the proliferation and invasion ability of hepatoma cells.

Objective To investigate the effect and mechanism of long noncoding RNA (lncRNA) PTEN pseudogene 1 (PTENP1) on the proliferation and migration of hepatocellular carcinoma (HCC) cells. Methods Lentiviral vectors expressing PTENP1 were constructed. HCC cells BEL-7404 were infected with LV003-GFP-PTENP1 and control vectors LV003-GFP. BEL-7404 cells stably expressing PTENP1 were constructed and the experimental and control groups were established. The proliferation and clone formation abilities of HCC cells in two groups were detected by CCK-8 assay and clonogenic assay. The migration ability of HCC cells was detected by wound healing assay. The expression of p44/42 mitogen-activated protein kinase (MAPK) and p38 MAPK proteins were detected by Western blot. Results The absorbance values A450 of the cells at 48 and 72 h in the experimental group were 1.4±0.3 and 2.3±1.1, signiifcantly lower compared with 3.2±1.7 and 3.4±1.1 in the control group (t=-5.78,-4.23;P<0.05). The number of cell clone formation in the experimental group was 55±12, signiifcantly less than 154±45 in the control group (t=-3.98, P<0.05). The percentage of cell migration in the experimental group was (21.7±2.6)%, signiifcantly lower than (57.7±4.9)%in the control group (t=-8.34, P<0.05). Western blot revealed that the expression of p44/42 MAPK and p38 MAPK proteins in the experimental group was significantly down-regulated compared with those in the control group. Conclusion lncRNA PTENP1 can inhibit the proliferation and migration of HCC cells probably through regulating MAPK signaling pathway.

Objective To investigate the effect and mechanism of liver cancer derived mesenchymal stem cell (LCMSC) on the invasion of liver cancer cells. Methods The expressions of interleukin (IL)-6, IL-8 and chemotactic factors CXCL1, CXCL5 and CXCL12 mRNA in the bone marrow derived mesenchymal stem cell (BMSC) and LCMSC were detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of protein CXCL12 in the supernate of LCMSC was detected by enzyme-linked immunosorbent assy (ELISA) and Western blot. Different cells were co-cultured and divided into the HepG2+BMSC, HepG2+LCMSC and HepG2+LCMSC+siRNA-CXCL12 groups. The effect of CXCL12 on the invasion of liver cancer HepG2 cells were detected by Transwell migration assay. The experiment data were compared using one way analysis of variance and LSD-t test or t test. Results The expression of CXCL12 mRNA in LCMSC was 60.3±2.4, significantly higher than 13.8±1.8 in BMSC (t=15.68, P<0.05). The expression of protein CXCL12 in the supernate of LCMSC was (31.5±1.7) ng/L, significantly higher than (14.3±1.5) ng/L in BMSC (t=7.60, P<0.05). And the expression of protein CXCL12 was up-regulated. Transwell migration assay indicated that the quantity of membrane-invasion cells in the HepG2+LCMSC group was 110±12, significantly higher than 65±9 in the HepG2+BMSC group and 76±7 in the HepG2+LCMSC+siRNA-CXCL12 group (LSD-t=5.25, 4.86; P<0.05). Conclusion CXCL12 is highly expressed in LCMSC. LCMSC may enhance the invasion of HepG2 cells through up-regulating the expression of CXCL12. The invasion of liver cancer cells can be effectively weakend by silencing the CXCL12 gene with siRNA.