Objective: To investigate the differences and diversity changes in gut microbiota between children and adolescents with constipation and diarrhea, and healthy individuals, and to explore the correlation between changes in stool consistency and gut microbiota, in order to provide a scientific reference for the research on intestinal microecology among children and adolescents. Methods: From October 2021 to March 2022, a total of 42 children and adolescents with constipation and 37 with diarrhea from a tertiary hospital in Hangzhou City, and 43 healthy individuals from 3 primary and secondary schools were included in this study. Fecal samples of children and adolescents were collected and then stool genomic DNA was extracted for 16S rRNA gene high throughput sequencing, and the sequencing results were analyzed. In the analysis of alpha diversity, the Kruskal-Wallis rank sum test was used to compare the differences between the three groups, and the FDR multiple testing correction was used for pairwise comparisons. In the analysis of beta diversity, the Adonis test was used to compare the overall differences between the three groups, and the ANOSIM test was used for pairwise comparisons. In the LEfSe analysis, the LDA scores obtained through LDA analysis (linear regression analysis). Results: Alpha diversity analysis showed that there were statistically significant differences in the Shannon index (4.01, 3.81, 4.19) and Simpson index (0.05, 0.06, 0.04) between the diarrhea group, constipation group, and healthy group ( H=6.05, 6.35, P <0.1). Further pairwise comparison showed that the Shannon index and Simpson index of the healthy group were higher than those of the constipation group ( P <0.1). Beta diversity analysis showed that the impact of grouping factors on inter group differences was statistically significant ( R 2=0.045, P <0.1). Community composition analysis showed that there were 234 species in total among the three groups, and 36 unique species in the healthy group, 36 species in the diarrhea group, and 48 species in the constipation group. Species difference analysis showed significant differences in species composition at the genus level among the three groups ( H=0.000 05, 0.000 16, 0.000 20, 0.000 21, 0.000 53, 0.001 39, P <0.1), including Lachnospiraceae of Firmicutes phylum, Eubacterium hallii, Veillonellaceae, Qscillospiraceae, Butyricicoccaceae and Staphylococcaceae, respectively. KEGG abundance statistics and COG functional analysis showed that there were no significant differences in gene expression abundance of the same function among the three groups ( P >0.1). Conclusions The different stool consistency of children and adolescents is related to changes in gut microbiota composition. Compared to the healthy group, children with constipation or diarrhea have disrupted gut microbiota balance, with a shift in dominant bacteria and a higher abundance of opportunistic pathogens.

In fixed prosthodontics, clear exposure of the preparation margin is the prerequisite for obtaining accurate digital impressions and improving the marginal fit of restorations. To resolve the issues associated with the cord retraction technique, such as pain, acute injury, and prolonged procedural time, this study proposes a new technology for intraoral digital impression taking with pneumatic gingival retraction. The new scanning head blows a high-speed airflow that instantaneously separates the free gingiva, locally exposing the subgingival preparation margin. Combined with the farthest point preservation stitching algorithm based on the distance from the normal vector and high-speed laser scanning photography, it achieves global preparation edge data and gingival reconstruction, realizing painless, non-invasive, and efficient precise acquisition of the preparation margin. Using this new technique, a patient with a full porcelain crown restoration on a posterior tooth was treated. The digital impression revealed a clear margin of the preparation, and the crown made from this data has a good marginal fit.

Objective:To investigate the incidence and treatment of gastric cancer in 16 medical centers in Tianjin from 2020 to 2021.Methods:The retrospective and descriptive study was conducted. The clinical data of 3 122 gastric cancer patients who underwent surgery in 16 medical centers, including Tianjin Medical University Cancer Institute & Hospital, et al, in Tianjin from 2020 to 2021 were collected. There were 2 112 males and 1 010 females, aged (64±11)years. Observation indicators: (1) general data of patients; (2) treatment situations; (3) postoperative complications. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(range). Count data were descri-bed as absolute numbers or percentages, and comparison between groups was conducted by the chi-square test. Results:(1) General data of patients. From 2020 to 2021, a total of 3 122 gastric cancer patients received surgeries in 16 medical centers in Tianjin, including 2 112 males and 1 010 females. There were 1 443 cases in 2020, including 976 males and 467 females, aged (63±11) years. There were 1 679 cases in 2021, including 1 136 males and 543 females, aged (65±11) years. Of the 3 122 pati-ents, cases in stage Ⅰ, Ⅱ, Ⅲ, Ⅳ were 696, 667, 1 466, 293, accounting for 22.293%(696/3 122), 21.365%(667/3 122), 46.957%(1 466/3 122), 9.385%(293/3 122), respectively. Cases with early gastric cancer, locally advanced gastric cancer, advanced gastric cancer account for 17.265%(539/3 122), 73.350%(2 290/3 122), 9.385%(293/3 122). There were 2 829 patients without distant metastasis and 293 patients with distant metastasis. For the 2 829 patients without distant metas-tasis, cases in stage T1, T2, T3, T4a, T4b accounted for 19.053%(539/2 829), 12.089%(342/2 829), 20.148%(570/2 829), 41.499%(1 174/2 829), 7.211%(204/2 829)respectively, cases in stage N0, N1, N2, N3 account for 37.328%(1 056/2 829), 16.331%(462/2 829), 15.836%(448/2 829), 30.505%(863/2 829). For the 293 advanced gastric cancer patients with distant metastasis, 190 cases had peri-toneal metastasis, 47 cases had lymph node metastasis, 27 cases had ovarian metastasis, 37 cases had liver metastasis, 14 cases had other metastasis (some patients had ≥2 distant metastases). (2) Treatment situations. ① For the 539 with early gastric cancer, cases undergoing endoscopic submu-cosal dissection, laparoscopic surgery, open surgery were 22, 150, 86 in 2020, versus 19, 212, 50 in 2021, showing a significant difference between them ( χ2=19.42, P<0.05). For the 498 patients with early gastric cancer who underwent laparoscopic or open surgery, cases undergoing open surgery including total gastrectomy, distal gastrectomy, proximal gastrectomy were 25, 81, 30, and cases undergoing laparoscopic surgery including total gastrectomy, distal gastrectomy, proximal gastrec-tomy were 18, 309, 35, respectively, showing a significant difference between them ( χ2=40.62, P<0.05). For the 2 290 patients with locally advanced gastric cancer, cases undergoing open surgery and laparoscopic surgery were 446 and 617 in 2020, versus 410 and 817 in 2021, showing a significant difference between them ( χ2=17.75, P<0.05). For the 2 290 patients with locally advanced gastric cancer, cases undergoing open surgery including total gastrectomy, distal gastrectomy, proxi-mal gastrectomy were 336, 377, 143, and cases undergoing laparoscopic surgery including total gastrectomy, distal gastrectomy, proximal gastrectomy were 377, 920, 137, respectively, showing a significant difference between them ( χ2=89.64, P<0.05). Of the 293 patients with advanced gastric cancer, 175 cases underwent surgeries due to hemorrhage, stenosis, perforation, 76 cases under-went surgery after chemotherapy, 42 cases underwent surgery directly. ② For 756 cases of 3 122 pati-ents undergoing total gastrectomy, 357 and 4 cases received open digestive tract reconstruction including Roux-en-Y and other anastomosis, versus 380 and 15 cases with laparoscopic digestive tract reconstruction including Roux-en-Y and other anastomosis, showing a significant difference between them ( χ2=5.57, P<0.05). For 1 687 cases undergoing distal gastrectomy, 84, 160, 158, 55 cases received open digestive tract reconstruction including Billroth Ⅰ anastomosis, Billroth Ⅱ + Braun anastomosis, Roux-en-Y anastomosis, uncut Roux-en-Y anastomosis, versus 154, 489, 417, 170 cases with laparoscopic digestive tract reconstruction including Billroth Ⅰ anastomosis, Billroth Ⅱ + Braun anastomosis, Roux-en-Y anastomosis, uncut Roux-en-Y anastomosis, showing a significant difference between them ( χ2=10.90, P<0.05) . Of the 539 patients with early gastric cancer, 65 cases had lymph node metastasis, in which 18 of 306 stage T1a cases had lymph node metastasis and 47 of 233 stage T1b cases had lymph node metastasis. The number of detected lymph nodes for the 2 290 patients with advanced gastric cancer was 31±15, including ≥16 for 2 059 cases and ≥30 for 1 276 cases. Of the 3 122 patients, cases with neoadjuvant therapy, complete response and incomplete response was 128, 13 and 115 in 2020, versus 250, 49 and 201 in 2021, showing a significant difference between them ( χ2=5.51, P<0.05). (3) Postoperative complications. Of the 3 122 patients, 746 cases had postoperative complications, with an incidence of 23.895%(746/3 122). There were 62 patients with grade 3 or more complications. Reoperation was conducted in 34 patients. There were 14 cases of postoperative death. The duration of postoperative hospital stay and hospital expense were (11±5)days and (98 114±46 598)yuan for the 3 122 patients, (26±14)days and (122 066±68 317)yuan for cases with complications, (40±21)days and (196 926±12 747)yuan for cases with grade 3 or more complications. Conclusion:Compared with 2020, cases undergoing laparoscopic surgery and distal gastrectomy for gastric cancer in Tianjin increases in 2021, and the digestive tract reconstruction also differs. The number of patients with neoadjuvant chemotherapy and complete response rate for advanced gastric cancer increases.

Objective:To evaluate the minimally invasive fixation with a locking plate via the tarsal sinus approach in the surgical treatment of Sanders Ⅱ and Ⅲ calcaneal fractures.Methods:A retrospective study was conducted to analyze the 65 patients who had been surgically treated for Sanders Ⅱ and Ⅲ calcaneal fractures at Department of Foot and Ankle Surgery, Binzhou Medical College Hospital from April, 2019 to September, 2020. There are 44 males and 21 females with an age of (42.5±10.4) years, and 46 Sanders type Ⅱ fractures and 19 Sanders type Ⅲ ones. The patients were divided into group L and group S according to surgical methods. Group S of 35 cases was fixated with a minimally invasive locking plate through the tarsal sinus incision while group L of 30 cases fixated with a locking plate through the L-shaped incision. The 2 groups were compared in terms of waiting time before surgery, surgical time, hospital stay, intraoperative bleeding, visual analogue scale (VAS) at 3 days after surgery, American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score at 1 year after surgery, excellent and good rate by the Maryland foot function score, B?hler angle, Gissane angle, varus angle, and complications.Results:There was no significant difference in the general data before surgery between the 2 groups, showing comparability ( P>0.05). All patients were followed up for (13.3±1.6) months after surgery. The waiting time before surgery [(2.8±1.8) d], surgical time [(80.7±9.9) min], hospital stay [(6.7±1.9) d], intraoperative bleeding [(54.3±14.2) mL], and VAS at 3 days after surgery [6 (5, 7) points] in group S were all significantly less or lower than those in group L [(8.2±2.8) d, (105.0±15.7) min, (14.6±3.4) d, (74.3±12.8) mL, and 7 (6, 8) points] (all P<0.05). At one year after surgery, the AOFAS ankle-hindfoot score [(90.1±3.5) points] in group S was significantly higher than that in group L [(83.5±6.7) points] ( P<0.05), but there was no statistically significant difference in the excellent and good rate by the Maryland foot function score between the 2 groups [91.4% (32/35) versus 86.7% (26/30)] ( P=0.695). The B?hler angle, Gissane angle, and varus angle were significantly improved in all patients one year after surgery compared with the values before surgery ( P<0.05), but there were no statistically significant differences within either group or between the 2 groups at 3 days or 1 year after surgery ( P>0.05). Peroneal muscle pain was reported in 1 case in group S; there were 2 cases of skin necrosis, 1 case of incision hematoma and 1 case of sural nerve injury in group L. Conclusion:The minimally invasive fixation with a locking plate via the tarsal sinus approach is an effective treatment for Sanders Ⅱ and Ⅲ calcaneal fractures.

OBJECTIVE: To measure the concentration of bone morphogenetic protein 2 (BMP-2) in demineralized bone matrix (DBM) prepared from different long bones and to evaluate the osteoinductivity of different DBM on MC3T3-E1 cells. METHODS: Different bones from the same cadaver donor were used as the initial materials for making DBM, which were divided into ulna group (uDBM), humerus group (hDBM), tibia group (tDBM), and femur group (fDBM) according to the origins, and boiled DBM (cDBM) was taken as the control group. The proteins of DBM were extracted by guanidine hydrochloride, and the concentrations of BMP-2 were determined by ELISA assay. Then the DBM were co-cultured with MC3T3-E1 cells, the proliferation of MC3T3-E1 cells was observed by cell counting kit 8 (CCK-8) assay. The osteogenic differentiation ability of MC3T3-E1 cells was qualitatively observed by alizarin red, alkaline phosphatase (ALP), and Van Gieson staining, and the osteogenic differentiation ability of MC3T3-E1 cells was quantitatively analyzed by ALP content. Linear regression was used to analyze the effect of BMP-2 concentration in DBM on ALP synthesis. RESULTS: There were significant differences in the concentration of BMP-2 among the DBM groups (P<0.05). The concentrations of BMP-2 in the lower limb long bone were higher than those in the upper limb long bone, and the concentration of BMP-2 in the fDBM group was about 35.5 times that in the uDBM group. CCK-8 assay showed that the cells in each group continued to proliferate within 5 days of co-culture, and the absorbance (A) values at different time points were in the order of cDBM groupP<0.05). Linear regression analysis showed that y^=0.361+0.017x, the effect of BMP-2 concentration in DBM on cellular ALP content was significant (t=3.552, P=0.005); for every 1 ng/g increase in BMP-2 concentration, ALP content would increase by 0.017 [95%CI (0.006, 0.027)] U/100 mL. CONCLUSION The concentration of natural BMP-2 in different long bones varies greatly, and the lower limb long bone is higher than the upper limb long bone. The harvested location of bone material was an important factor affecting the osteoinductivity of DBM.
Alkaline Phosphatase ; Bone Matrix ; Bone Morphogenetic Protein 2 ; Cell Count ; Coloring Agents ; Osteogenesis ; Animals ; Mice

OBJECTIVE: To study the effectiveness of TightRope elastic fixation combined with functional total repair of the inferior tibiofibular ligament in the treatment of distal tibiofibular syndesmosis injury. METHODS: The clinical data of 34 patients with distal tibiofibular syndesmosis injury who met the selection criteria between January 2020 and January 2022 were retrospectively analyzed, and they were divided into improved group (TightRope elastic fixation combined with functional total repair of inferior tibiofibular ligament) and control group (distal tibiofibular screw fixation) according to the surgical methods, with 17 cases in each group. There was no significant difference in age, gender, body mass index, fracture type, and other baseline data between the two groups (P>0.05). The operation time, intraoperative blood loss, and complications were recorded in the two groups. The American Orthopaedic Foot and Ankle Society (AOFAS) score, ankle metatarsal flexion and dorsal extension range of motion were used to evaluate the ankle function. The patient satisfaction survey was conducted at last follow-up. RESULTS: All 34 patients were followed up 8-20 months, with a median of 13 months. The operation time and intraoperative blood loss in the improved group were significantly longer than that in the control group (P<0.05). In the improved group, no infection or poor reduction occurred, and only 1 patient had TightRope knot reaction at 6 months after operation. In the control group, there were 2 cases of poor reduction, 1 case of lower tibiofibular screw rupture, and 1 case of subcutaneous infection (cured after anti-infection treatment). There was no significant difference in the incidence of complications between the two groups (P>0.05). At last follow-up, the AOFAS score and ankle metatarsal flexion and dorsal extension range of motion of the improved group were significantly better than those of the control group (P<0.05). The satisfaction rates of patients in the improved group and the control group were 94.1% and 82.4%, respectively, showing significant difference (P<0.05). CONCLUSION TightRope elastic fixation combined with functional total repair of inferior tibiofibular ligament in the treatment of distal tibiofibular syndesmosis injury has sufficient fixation strength, and can achieve better effectiveness and joint function compared with traditional screw fixation.
Humans ; Ankle Joint/surgery* ; Blood Loss, Surgical ; Ligaments/surgery* ; Plastic Surgery Procedures ; Retrospective Studies ; Ankle Injuries/surgery*

OBJECTIVE: To compare the effectiveness of subtalar arthroereisis (STA) combined with modified Kidner procedure versus STA alone in the treatment of flexible flatfoot combined with painful accessory navicular bone in children. METHODS: The clinical data of 33 children with flexible flatfoot combined with painful accessory navicular bone who were admitted between August 2018 and August 2021 and met the selection criteria were retrospectively analyzed. They were divided into a combination group (17 cases, treated by STA combined with modified Kidner procedure) and a control group (16 cases, treated by STA alone) according to the surgical methods. There was no significant difference in baseline data between the two groups ( P>0.05), such as gender, age, affected side of the foot, disease duration, and preoperative visual analogue scale (VAS) score, American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score, talus-first metatarsal angle (T1MT), talus-second metatarsal angle (T2MT), talonavicular coverage angle (TCA), talus first plantar angle (Meary angle), calcaneal inclination angle (Pitch angle), and heel valgus angle (HV). The operation time, incision length, intraoperative blood loss, number of intraoperative fluoroscopies, and perioperative complications were recorded in both groups. The anteroposterior, lateral, and calcaneal axial X-ray films for the affected feet were taken regularly, and T1MT, T2MT, TCA, Meary angle, Pitch angle, and HV were measured. The VAS score, AOFAS ankle-hindfoot score were used to evaluate pain and functional recovery before and after operation. RESULTS: Surgeries in both groups were successfully performed without surgical complication such as vascular, nerve, or tendon injuries. Less operation time, shorter incision length, less intraoperative blood loss, and fewer intraoperative fluoroscopies were found in the control group than in the combination group ( P<0.05). One case in the combination group had partial necrosis of the skin at the edge of the incision, which healed after the dressing change and infrared light therapy, and the rest of the incisions healed by first intention. All children were followed up 12-36 months, with a mean of 19.6 months. At last follow-up, VAS score and AOFAS ankle-hindfoot score significantly improved in both groups when compared with preoperative ones ( P<0.05), and the differences of these scores between before and after operation improved more significantly in the combination group than in the control group ( P<0.05). Imaging results showed that the T1MT, T2MT, TCA, Meary angle, and HV significantly improved in both groups at last follow-up when compared with preoperative ones ( P<0.05), and the Pitch angle had no significant difference when compared with preoperative one ( P>0.05). But there was no significant difference in the difference of these indicators between before and after operation between the two groups ( P>0.05). CONCLUSION Both procedures are effective in the treatment of flexible flatfoot children with painful accessory navicular bone. STA has the advantage of minimally invasive, while STA combined with modified Kidner procedure has better effectiveness.
Humans ; Child ; Flatfoot/surgery* ; Blood Loss, Surgical ; Retrospective Studies ; Treatment Outcome ; Osteotomy/methods* ; Talus ; Pain

Objective    To study the effect of Tangeretin on non-small cell lung cancer (NSCLC) and the tumor stemness, and to find the molecular mechanism of its effect. Methods    We used cell counting and cell cloning experiments to study the effect of Tangeretin on the proliferation of NSCLC cells in vitro. The effect of Tangeretin on the invasion of NSCLC cells was detected by transwell assay. We detected the effect of Tangeretin on the proliferation of NSCLC cells in vivo by nude mouse tumor-bearing experiment. The effect of Tangeretin on tumor stemness of NSCLC cells was detected by self-renew assay, and CD133 and Nanog protein expressions. The expressions of PI3K/AKT/mTOR signaling pathway-related proteins were detected by Western blotting (WB). Results    Tangeretin had a good inhibitory effect on the proliferation of NSCLC cells in vivo and in vitro. Cell counting experiment, clonal formation experiment and nude mouse tumor-bearing experiment showed that Tangeretin could inhibit the proliferation activity, clonal formation ability, and tumor size of NSCLC cells in vivo. Self-renew experiments showed that Tangeretin could inhibit the self-renew ability of NSCLC cells. WB experiments showed that Tangeretin inhibited the expressions of tumor stemness markers CD133 and Nanog in NSCLC cells. Tangeretin could inhibit the activation of PI3K/AKT/mTOR signaling pathway-related proteins in NSCLC cells, and the activation of PI3K/AKT/mTOR signaling pathway could partially remit the inhibitory  effect of Tangeretin on tumor stemness of NSCLC cells. Conclusion    Tangeretin can inhibit the tumor stemness of NSCLC cells, which may be related to the regulation of PI3K/AKT/mTOR signaling pathway.

Objective    To investigate the effects of telmisartan on the proliferation, migration and apoptosis of non-small cell lung cancer A549 and the mechanism of regulating Wnt signaling pathway. Methods    Non-small cell lung cancer cell line A549 was cultured in vitro. Cell counting kit-8 (CCK-8) assay was used to detect the effect of telmisartan at different concentrations on the proliferative activity of A549 cells. The survival fraction of A549 treated with different concentrations of telmisartan was determined by colony-formation assay. The effect of telmisartan at different concentrations on the migration ability of A549 cells was examined in the wounding healing assay. Hoechst staining was used to detect the effects of telmisartan at different concentrations on the apoptosis of A549. Western bloting was used to detect the expressions of β-actin, proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, Wnt-3a, Beta-catenin (β-catenin), serine protein kinase 3β (p-GSK-3β), glycogen synthase kinase-3β (GSK-3β) and c-myc. Results    Different concentrations of telmisartan treatment inhibited the proliferation activity, colony-formation rate and migration of A549 cells, and reduced the expression of PCNA in a concentration-dependent manner. Telmisartan treatment promoted the apoptosis of A549 cells, significantly increased the expression of pro-apoptotic protein Bax and decreased the expression of anti-apoptotic protein Bcl-2. The expression levels of Wnt-3a, β-catenin, p-GSK-3β, and c-myc in A549 cells increased after treatment with telmisartan, while the expression levels of GSK-3β decreased. Conclusion    Telmisartan may play a role in the proliferation, migration and apoptosis of non-small cell lung cancer A549 cells, and  inhibiting the Wnt/β-catenin signaling pathway may be one of the mechanisms.

To explore the different chloroplast genome characteristics of Sinopodophyllum hexandrum, five chloroplast genome sequences of S. hexandrum were compared. Its genome map, repeat sequence, codon preference, inverted repeat (IR)/single-copy (SC) boundary, alignment of chloroplast genome sequences and phylogenetic were analyzed using bioinformatics tools. The results showed that: the total length of five chloroplast genomes of S. hexandrum, with a typical tetrad structure, were 157 203-157 940 bp, and a total of 133-137 genes were annotated, reflecting the diversity of chloroplast genomes of S. hexandrum. Different chloroplast genomes of S. hexandrum has different simple sequence repeat (SSR), where simple repeat of single nucleotide of A/T were the majority among the SSR detected. The interspersed repetitive sequences included direct, palindromic and inverted repeats. The value of effective number of codon (ENc) which was analyzed by using codon bias was 51.14~51.17, the proportion of GC and GC3s was less than 50%, the codon usage pattern tended towards frequently use of A/U-ending bases. Genome sequences and the IR/SC boundaries of five chloroplast genomes of S. hexandrum were relatively conservative. Phylogenetic analysis showed that S. hexandrum and Podophyllum pettatum had the closest genetic relationship. In summary, the chloroplast genome characteristics and evolutionary relationship of different chloroplast genomes of S. hexandrum were obtained, which may facilitate the utilization, protection, variety identification and genetic evolution of S. hexandrum resources.
Phylogeny ; Genome, Chloroplast ; Chloroplasts/genetics* ; Genomics ; Evolution, Molecular