Microbial communities such as those residing in the human gut are highly diverse and complex, and many with important implications for health and diseases. The effects and functions of these microbial communities are determined not only by their species compositions and diversities but also by the dynamic intra- and inter-cellular states at the transcriptional level. Powerful and scalable technologies capable of acquiring single-microbe-resolution RNA sequencing information in order to achieve a comprehensive understanding of complex microbial communities together with their hosts are therefore utterly needed. Here we report the development and utilization of a droplet-based smRNA-seq (single-microbe RNA sequencing) method capable of identifying large species varieties in human samples, which we name smRandom-seq2. Together with a triple-module computational pipeline designed for the bacteria and bacteriophage sequencing data by smRandom-seq2 in four human gut samples, we established a single-cell level bacterial transcriptional landscape of human gut microbiome, which included 29,742 single microbes and 329 unique species. Distinct adaptive response states among species in Prevotella and Roseburia genera and intrinsic adaptive strategy heterogeneity in Phascolarctobacterium succinatutens were uncovered. Additionally, we identified hundreds of novel host-phage transcriptional activity associations in the human gut microbiome. Our results indicated that smRandom-seq2 is a high-throughput and high-resolution smRNA-seq technique that is highly adaptable to complex microbial communities in real-world situations and promises new perspectives in the understanding of human microbiomes.
Humans ; Gastrointestinal Microbiome/genetics* ; Bacteriophages/physiology* ; High-Throughput Nucleotide Sequencing ; Sequence Analysis, RNA/methods* ; Bacteria/virology*

As people's living standards improve， the development trend of diabetes has gradually become severe. Diabetes is a chronic inflammatory disease associated with abnormal expression of nuclear factor-kappa B （NF-κB） in patients. NF-κB exists in various tissue cells and participates in the regulation of a variety of genes related to immune function and inflammation. Varieties of factors can activate NF-κB when the body is stimulated by external factors， so as to produce inflammation and other reactions. Previous studies on NF-κB mainly focus on cancer， and the pathological mechanism of the treatment of diabetes by related signaling pathways and the progress of traditional Chinese medicine （TCM） treatment have not been systematically elaborated on. By referring to the relevant literature in China and abroad， it was found that NF-κB is not isolated in the development and progression of diabetes but is associated with signal molecules related to inflammation， oxidative stress， and energy metabolism， and it is involved in mediating inflammation， pancreatic β cell apoptosis， insulin signal transduction， and other physiological functions. Therefore， blocking the transmission of NF-κB signaling pathway is beneficial to the treatment of diabetes. At present， Western medicine for the treatment of diabetes mainly includes oral hypoglycemic drugs and insulin injections， but the adverse reactions are obvious. TCM has been characterized by multi-target， extensive action， and excellent curative effects in the treatment of diabetes. TCM and its compounds with functions of tonifying Qi and promoting blood circulation， regulating qi and eliminating phlegm， clearing heat and detoxifying， and nourishing Yin and moistening dryness can effectively intervene in the abnormal expression of NF-κB signaling pathway in vivo through anti-inflammatory effects. In this paper， the association between NF-κB signaling pathway and diabetes was summarized， and the modern research progress of TCM intervention of NF-κB signaling pathway in the treatment of diabetes in the past five years was reviewed， so as to lay a laboratory foundation for the study of a new pathological mechanism of diabetes based on NF-κB signaling pathway and provide new targets and research direction for the prevention and treatment of diabetes and development of related TCM.

Objective:To delineate the surgical methodology and therapeutic paradigm of proximal tibial notch retrograde interlocking intramedullary nailing for ameliorating deformities due to osteofibrous dysplasia (OFD) in a pediatric population.Methods:A retrospective assessment was conducted on the medical records of individuals undergoing orthopedic osteotomy complemented by retrograde interlocking intramedullary nailing for OFD of the tibia from January 2016 to December 2019. The cohort comprised 15 patients, with a follow-up exceeding three years, documenting complete data sets. The patient profile included 8 males and 7 females, with 8 left-side and 7 right-side afflictions. The mean age at the time of surgery was 10.1±2.5 years, ranging from 7.1 to 12.6 years. Parameters measured were preoperative and postoperative imaging findings, which encompassed the scope of the lesion (longitudinal lesion length relative to tibial length), coronal and sagittal limb alignments, and lower limb length discrepancies.Results:The mean follow-up duration was 3.4±1.3 years, ranging from 3 to 6.6 years. Preoperatively, prominent anterior tibial arch deformities and limping were present, with 7 cases reporting fatigue-induced pain and 3 instances of pathological fractures. Post-surgery, pain symptoms were resolved, gait disturbances were improved in 9 patients, and completely resolved in 6. Tibial osteotomy or fracture healing of 15 patients averaged 3.9±0.7 months (range 3-5 months). The lesion range before surgery was 0.41±0.17, immediately after surgery was 0.38±0.17, and at the last follow-up was 0.30±0.16, with no statistical significance ( F=0.101, P=0.904). Lesion range showed no significant change throughout treatment, but radiographic density within the lesion notably increased post-surgery, suggesting bone improvement. The anterior tibial arch Angle was 28.30°±6.62° (range 20°-45°) before surgery, 4.73°±1.53° immediately after surgery, and 6.87°±1.36° at the last follow-up, with statistical significance ( F=159.739, P<0.001). A significant correction in the anterior tibial arch deformity was achieved and maintained postoperatively. There was no significant angular deformity of the tibia in the coronal plane before operation, and the medial proximal tibial angle (MPTA) and lateral distal tibial angle (LDTA) were 87.50°±1.46° and 88.30°±1.62°, 88.40°±1.46° and 88.70°±1.45° immediately after surgery, and 88.00°±1.39° and 89.10°±1.53° at the last follow-up, the differences were statistically significant ( F=1.741, P=0.188; F=1.016, P=0.371), there was no coronal deformity of tibia. The limb length discrepancy (LLD) was 0.60±0.98 cm before surgery, 0.18±0.93 cm at the last follow-up, with statistical significance ( t=0.096, P=0.761). There were no incidents of postoperative complications such as infection. Conclusion:In pediatric cases of tibial deformities attributed to osteofibrous dysplasia, a therapeutic strategy involving osteotomy for lower limb realignment, sans curettage or bone grafting of the lesion, followed by retrograde interlocking intramedullary nailing, yields favorable outcomes. Importantly, this implantation technique does not compromise the integrity of the proximal tibial epiphyseal plate in children and adolescents.

Objective:To explore the clinical effect of plating after lengthening to assist the consolidation of tibial shortening deformity in children.Methods:A retrospective analysis was conducted on 10 children with tibial shortening who were treated with circular external fixator lengthening and replacement with plate-assisted internal fixation (study group) in the Department of Pediatric Orthopedics of Tianjin Hospital from November 2019 to October 2022, and 16 children who were treated by circular external fixator only during the same period were enrolled as the external fixator group (control group). Among the 26 cases, there were 15 males and 11 females, 10 left knees and 16 right knees. The average age at the time of surgery was 8.8±4.7 years (range 5.1-13.8 years). The gender, side, age at the time of surgery were compared between the two groups. The extension length, external fixator index, healing index, Kolcaba comfort scale score, knee and ankle joint range of motion, complications and average total hospitalization costs were recorded.Results:There were no significant differences in gender, side, age at the time of surgery between the study group and control group. The extended lengths were 5.44±1.25 cm and 5.78±1.11 cm respectively without significant difference ( t=0.096, P=0.096). The external fixator index and healing index were 17.86±2.94 d/cm vs. 50.97±7.03 d/cm and 40.94±6.63 d/cm vs. 45.24±5.98 d/cm in study group and the control group with significant differences ( t=13.299, P<0.001; t=1.289, P=0.033). The Kolcaba comfort scale score of the children in the study group was higher than that of the control group, with significant difference ( t=6.821, P=0.001). Comparing the range of motion of the knee and ankle joints between the two groups, there were no significant differences before surgery, at the end of extension surgery, and at the final follow-up. When the extension end was healed, the range of motion of the knee joint was137.89°±4.40° vs. 114.09°±13.60° and ankle joint was 64.35°±5.50° vs. 56.65°±8.86° in the study group and control group with significant difference ( t=17.235, P<0.001; t=7.821, P=0.002). In the study group, 4 cases had pin tract infection, but no refracture occurred; in the control group, pin tract infection occurred in 13 cases, and refracture occurred in 2 cases after removal of the external fixator. The average total hospitalization cost of the study group was higher than that of the control group with significant difference ( t=3.745, P=0.036). Conclusion:The clinical effect of replacing plate-assisted internal fixator during the mineralization period of tibial shortening in children is reliable, and can significantly shorten the time for using external fixator. It is beneficial to the healing of the extended end of the osteotomy and the functional recovery of the knee and ankle joints. Being more comfortable for children with fewer complications, it is applicable for children who need long-distance extension, multiple extensions with limited joint function, poor tolerance for external fixators and low treatment compliance.

This report described one patient of giant polycystic liver and polycystic kidney involving iliac fossa. Preoperative computed tomography (CT) revealed a large polycystic kidney occupying partially iliac fossa space. A decompression of lower pole of original kidney was planned for placing transplanted kidney. During total liver resection plus orthotopic liver transplantation, right polycystic kidney could move up on its own and iliac fossa space was released for placing transplanted kidney smoothly. Polycystic kidney shrunk markedly post-operation. It provided references for surgical planning of combined liver-kidney transplantation for this type of disease.

ObjectiveTo explore the clinical efficacy of Gouteng prescription in treating the patients with primary hypertension with anxiety disorder due to yang hyperactivity and heat toxin and the impact of the formula on the balance of inflammatory cytokines. MethodA total of 98 patients diagnosed with primary hypertension and anxiety disorder were randomized into control and observation groups. On the basis of conventional western medicine treatment for hypertension， the control group （47 patients） was treated with Shugan Jieyu capsules for 8 weeks， while the treatment group （51 patients） with Gouteng prescription for 8 weeks. The two groups were compared in terms of the blood pressure level， 24-hour blood pressure variability， Hamilton anxiety scale （HAMA） score， Pittsburgh sleep quality index （PSQI） score， quality of life （SF-36 scale） score， traditional Chinese medicine （TCM） syndrome score and efficacy， incidence of adverse reactions， and the levels of interleukin （IL）-1β， IL-6， IL-10， and IL-4 in the serum of peripheral blood. ResultThe final trial was completed with 95 patients， including 46 in the control group and 49 in the observation group. The treatment in both groups lowered the blood pressure and blood pressure variability （P<0.05， P<0.01）. The observation group outperformed the control group in recovering the systolic blood pressure （SBP）， 24-hour mean systolic blood pressure （24 h SBP）， 24-hour systolic blood pressure variability （24 h SBPV）， and 24-hour diastolic blood pressure variability （24 h DBPV） （P<0.05）. After treatment， the HAMA and PSQI scores in both groups decreased （P<0.05， P<0.01）， and the observation group had lower HAMA and PSQI scores than the control group （P<0.05）. Compared with those before treatment， the SF-36 scores in both groups increased （P<0.05， P<0.01）. After treatment， the observation group had higher scores of physiological function （PF）， bodily pain （BP）， social function （SF）， role-emotional （RE）， and mental health （MH） indicators than the control group （P<0.05）. After treatment， the TCM syndrome scores in both groups decreased （P<0.05， P<0.01）， and the observation group had lower score than the control group （P<0.05）. The total response rate regarding TCM syndrome in the observation group was 85.71% （42/49）， which was higher than that （63.04%， 29/46） in the control group （χ²=6.621， P<0.05）. The treatment in both groups lowered the levels of pro-inflammatory cytokines （IL-1β， IL-6） and elevated the levels of anti-inflammatory cytokines （IL-10， IL-4） （P<0.05， P<0.01）， and the changes were more obvious in the observation group than in the control group （P<0.05）. There were no adverse events during the research process. ConclusionGouteng prescription can recover the blood pressure level， reduce blood pressure variability， suppress anxiety state， improve sleep and quality of life， decrease TCM syndrome score， increase total response rate， lower serum IL-1β and IL-6 levels， and elevate serum IL-10 and IL-4 levels in the patients with primary hypertension complicated with anxiety disorder due to yang hyperactivity and heat toxin. It may exert the effects by regulating the balance of pro-inflammatory and anti-inflammatory cytokines.

ObjectiveTo investigate the effect of rutin on the browning of 3T3-L1 preadipocytes and the mechanism. MethodCell counting kit-8 （CCK-8） assay was used to detect the effect of different concentration of rutin （3.125， 6.25， 12.5， 25， 50， 100， 200 μmol·L^-1） on 3T3-L1 cell activity， and Western blot to examine the effect of rutin （12.5， 25， 50 μmol·L^-1） on the expression of thermogenesis-associated proteins uncoupling protein 1 （UCP1）， PR domain containing 16 （PRDM16） and peroxisome proliferator-activated receptor γ coactivator-1α （PGC-1α） in adipocytes. After the optimal concentration of rutin was determined， the effect of rutin on lipid droplet formation in adipocytes was observed based on oil red O staining， and the expression of nuclear respiratory factor 1 （NRF1）， nuclear respiratory factor 2 （NRF2） and mitochondrial transcription factor A （TFAM）， which were the landmark proteins of mitochondrial biosynthesis， was detected by Western blot. ResultCompared with the blank group， 200 μmol·L^-1 rutin inhibited 3T3-L1 cell activity （P<0.01）. Compared with the blank group， at the concentration of 12.5， 25， 50 μmol·L^-1 rutin significantly promoted the expression of thermogenesis-associated proteins （UCP1， PRDM16， and PGC-1α） （P<0.01）， which was determined as the optimal concentration. Compared with the blank group， 50 μmol·L^-1 rutin significantly increased the immunofluorescence intensity of mitochondrial UCP1 protein in 3T3-L1 cells （P<0.01） and the expression of the markers of mitochondrial biosynthesis （NRF1， NRF2， and TFAM） （P<0.01）. In addition， 50 μmol·L^-1 rutin significantly inhibited lipid droplet formation of 3T3-L1 adipocytes （P<0.01）. ConclusionRutin inhibited lipid droplet deposition in 3T3-L1 adipocytes and increased the expression of thermogenesis-related proteins （UCP1， PRDM16， and PGC-1α） and markers of mitochondrial biosynthesis （NRF1， NRF2， and TFAM）， thereby inducing the browning of 3T3-L1 adipocytes. This lays a basis for the development of drugs that safely regulate the browning of white cells.

Objective:To design an anatomical plate of ulna coronoid process using 3D printing and computer model design software based on a collection of CT scanning data of the ulna coronoid process.Methods:The CT scans of the elbow joint with no obvious anatomic variation, no fracture, or no history of elbow operation were collected which had been taken at Trauma Center, The First Affiliated Hospital of Kunming Medical University from September 2017 to January 2022. There were 52 males and 50 females. RadiAnt DICOM Viewer and Mimics Medical 21.0 were used to visualize the CT data of the elbow joint of 102 volunteers. The software was used to measure the angle between the tip of the ulna coronoid process and the tuberosity of the ulna, the width at 1/2 height of the ulna coronoid process, the distance between the tip of the ulna coronoid process and the horizontal plane of the ulna tuberosity, and the safety angle for screw placement. After the values were measured, Siemens Ungraphics NX12.0 software was used to design the anatomical plate and the screw guide device of the ulna coronoid process. After the plate model was designed, a 1:1 actual plate model of the ulna coronoid process was produced by 3D printing. The actual plate model was placed onto an adult model of the ulna coronoid process and an adult cadaveric specimen of the ulna coronoid process to verify its matching degree. An in vitro operation was simulated using the plate model to verify its operability. Results:There were no significant differences between the left and right sides in the angle between the tip of the ulna coronoid process and the tuberosity of the ulna, the width at 1/2 height of the ulna coronoid process, the distance between the tip of the ulna coronoid process and the horizontal plane of the ulna tuberosity, or the safety angle for screw placement in either males or females ( P>0.05). There were no significant differences between males and females in the angle between the tip of the ulna coronoid process and the tuberosity of the ulna or in the safety angle for screw placement ( P>0.05). There were statistically significant differences between males and females in the width of 1/2 height of the ulna coronoid process and the distance between the tip of the ulna coronoid process and the horizontal plane of the ulna tuberosity ( P<0.05). However, the experiments on computer simulative adaptation and plate model simulative adaptation found that the anatomical plates of the ulna coronoid process designed on various parameters of males and females were exchangeable, leading to similarly good marching degrees and safe angles for screw placement. Conclusions:The anatomical plate of the ulna coronoid process designed in this study demonstrates a good fit and a safe angle for screw placement, basically achieving the goal expected to provide a basis for fabrication of a titanium alloy plate. In design of an anatomical plate of ulna coronoid process, it is not necessary to differentiate males from females or to differentiate the left side from the right one, because only a general plate can be used for both males and females and for both the left and the right sides.

Objective:To investigate the effect of fat mass index (FMI) on early recovery after total knee arthroplasty (TKA).Methods:Patients who underwent primary unilateral TKA in Xi'an Honghui Hospital from July 2020 to July 2021 were retrospectively analyzed. The preoperative body composition was measured by dual energy X-ray absorptiometry and the FMI was calculated. Patients were divided into normal group (male: 3.0-6.0 kg/m 2; female: 5.0-9.0 kg/m 2), overweight group (male: 6.1-9.0 kg/m 2; female: 9.1-13.0 kg/m 2), and obese group (male: >9 kg/m 2; female: >13 kg/m 2) according to level of FMI, and the operation time, blood loss, and incidence of postoperative complications were collected. Multifactorial analysis of the effect of FMI on early recovery after TKA was performed using a generalized linear model. Draw the receiver operating characteristics (ROC) curve of BMI and FMI on the predicted effect of postoperative Western Ontario and McMaster Universities (WOMAC) osteoarthritis index scores and Knee Society Score (KSS) to compare the effect of FMI with BMI on early recovery after TKA. Results:A total of 100 patients were included in the study, 24 males and 76 females, aged 65.0±8.2 years (range, 42-81 years). There were 15 cases in normal group, 55 cases in overweight group and 30 cases in obese group. All patients successfully completed the operation and were followed up for 3.15±0.72 months (range, 2.8-3.2 months). The WOMAC scores of the obese group at 2 weeks, 1 and 2 months postoperative were 34.57±3.68, 22.03±2.79, and 15.77±2.96, which were greater than those of the normal group (28.73 ±2.58, 19.07±2.71, 12.27±3.10), as well as the overweight group (30.05±4.09, 19.33±2.42, 14.84±2.42), with statistically significant differences ( P<0.05). The KSS scores of the obese group at postoperative 1 and 2 months were 68.83±5.52 and 81.17±4.49, which were lower than those of the normal group (77.33±5.63, 87.33±4.17), as well as the overweight group (72.64±5.43, 83.73 ±5.02), with statistically significant differences ( P<0.05). The WOMAC score, KSS score, and postoperative complications at 2 months postoperatively were selected as outcome indicators to plot the ROC curve, and the ROC curve for the WOMAC score at 2 months postoperatively showed an area under the curve corresponding to FMI of 0.744 (95% CI: 0.54, 0.82), which was greater than that of BMI [0.624 (95% CI: 0.51, 0.74)], and the difference was statistically significant ( Z=2.19, P=0.021). The ROC curve for the KSS score at 2 months postoperatively showed an area under the curve corresponding to FMI of 0.718 (95% CI: 0.62, 0.82), which was greater than that of BMI [0.612 (95% CI: 0.52, 0.74)], with a statistically significant difference ( Z=2.58, P=0.016). The ROC curve for postoperative complications showed an area under the curve of 0.639 (95% CI: 0.41, 0.88) for FMI and 0.605 (95% CI: 0.37, 0.84) for BMI, with no statistically significant difference ( Z=0.48, P=0.632). Conclusion:The greater the FMI the poorer the early functional recovery after initial TKA, and FMI is more valuable than BMI in predicting the early functional recovery.

Objective:To investigate the clinical effect of "ladder reduction method" in the treatment of iliac fracture combined anterior dislocation of sacroiliac joint.Methods:The retrospective analysis was performed on 10 cases of iliac fracture combined anterior sacroiliac joint dislocation admitted to the Affiliated Hospital of Yunnan University from February 2010 to January 2022, among which 5 cases were males and 5 cases were females, aged ranging from 22 to 52 years, with an average age of 38.8 years. All patients were injured in car accidents including 5 cases of C1.2, 3 cases of C2, and 2 cases of C3 fractures according to Tile classification. All patients were treated with the "ladder reduction method" with plate and screw fixation. In the first step, 1-2 Schanz pins were inserted into the iliac crest to control the ilium, and the Schanz pins were appropriately pulled laterally; in the second step, the periosteal stripper was used to pry the reduction between the sacrum and ilium; in the third step, for the patients who still could not be reduced, a 2.5 mm diameter Kirschner wire was placed on the sacrum close to the iliac crest, and a periosteal stripper was inserted between the sacrum and iliac crest, with its tip against the Kirkner wire, and the iliac crest as the fulcrum for pry pulling to separate the two. In the fourth step, the pry was maintained, and then another 2.5 mm diameter Kirschner wire was placed on the sacrum close to the internal margin of the iliac bone. The periosteal stripper was continued to pry between the sacrum and the iliac bone, and the operation was repeated. At the same time, the anterior dislocation of the sacroiliac joint was reduced with traction of the lower limb. Postoperatively, the quality of reduction was evaluated by the Matta score, and the degree of functional recovery after pelvic fracture was evaluated by the Majeed score.Results:Four patients completed the reduction through the first and second steps, and 6 cases of refractory sacroiliac joint anterior dislocation were successfully reduced through the first to fourth steps. The fracture reduction time of 6 patients with refractory anterior sacroiliac joint dislocation was 39.67±3.09 min (range, 35-44 min), with intraoperative blood loss of 300.00±141.42 ml (range, 150-600 ml); in the other 4 cases, the fracture reduction time was 36.75±4.38 min (range, 30-42 min), and the intraoperative blood loss was 225.00±44.30 ml (range, 200-300 ml). All 10 patients were followed up for 12.9±3.7 months (range, 9-20 months). The anterior and posterior pelvic ring fractures were healed in all patients, and the fracture healing time was 12.77±1.62 weeks (range, 10.71-15.28 weeks). At the last follow-up, Matta evaluation was excellent in 5 cases, good in 1 case, and excellent in the other 4 cases. The Majeed scores of 6 cases were 86.50±6.08 points (range, 74-92 points), of which 5 cases were excellent and 1 case was good. The other 4 cases were 81.5±9.39 scores (range, 71-94), of which 2 were excellent and 2 were good.Conclusion:The "ladder reduction method" is a safe, effective and easy-to-operate method for the treatment of iliac fracture combined anterior dislocation of the sacroiliac joint, especially for refractory anterior dislocation of the sacroiliac joint, which can still obtain satisfactory curative effects.