[Objective] To extract hemoglobin (Hb) from red blood cells using osmotic pressure swelling method, expected to achieve a hemoglobin dissolution rate of ≥80% and a cell membrane integrity rate of ≥70%. [Methods] Human umbilical cord blood red blood cells were used as raw materials and phosphate buffer solution was used as the swelling solution for red blood cells. A three factor three-level orthogonal experiment (n=3) was conducted to determine the optimal matching conditions for selecting the osmolality molar concentration of phosphate buffer solution, pH value of hypotonic phosphate buffer solution and volume ratio of hypotonic phosphate buffer solution to washed red blood cells. Red blood cell swelling solution samples (n=6) were prepared by the optimal matching conditions and the original process conditions. The hemoglobin dissolution rate and cell membrane integrity rate were checked. In the expanded comparative experiment, red blood cell swelling solution samples (n=6) were prepared by the optimal matching conditions and the original process conditions, which was filtered by ultrafiltration membranes. The filtration time and hemoglobin yield were checked. [Results] The optimal matching conditions for preparing red blood cell swelling solution were obtained through orthogonal experiment as follows: osmotic pressure molar concentration was 30 mOsmol/Kg, pH was 7.8, and phosphate buffer to red blood cell volume ratio was 6∶1. On the basis of the above conditions, the red blood cell swelling solution sample was compared with the original process sample: the hemoglobin dissolution rate was (82.4±1.8)% vs (78.6±3.0)% (P<0.05), and the cell membrane integrity rate was (65.8±4.0)% vs (28.7±2.3)% (P<0.05). In the expanded comparative experiment, the optimal matching conditions were compared with the original process conditions: filtration time(s) (327±9) vs (434±13) (P<0.05), and hemoglobin yield was (72.3±1.2)% vs (66.0±1.4)% (P<0.05). [Conclusion] Compared with the original preparation process, the hemoglobin extraction process which optimized through orthogonal experiments greatly reduces the cell membrane fragmentation rate and minimizes the entry of cell membrane matrix into the target solution, ensuring a slightly higher hemoglobin dissolution rate, and reducing the preparation difficulty for the subsequent cell membrane separation and further purification.

[Objective] To investigate the protective effect of Salvia miltiorrhiza injection (SMI) on the kidneys of HBOC-CHP01 resuscitated haemorrhagic shock rats. [Methods] A 50% haemorrhagic shock rat model was established, with 12 rats divided into two groups: SMI + HBOC-CHP01 group and HBOC-CHP01 group, with 6 rats in each group. The rats in the SMI+ HBOC-CHP01 group were given an equal volume of HBOC-CHP01 for resuscitation after haemorrhagic shock, and an 8 mL/kg dose of SMI. Rats in the HBOC-CHP01 group were resuscitated by administering an equilibrium blood loss volume of HBOC-CHP01 and given an 8 mL/kg dose of 0.9% NaCl solution. Blood was taken from rats at five points: before bloodletting (baseline), during haemorrhagic shock (HS), immediately after resuscitation (RS0h), 1 h after resuscitation (RS1h), and 24 h after resuscitation (RS24h). A blood gas analyser was used to detect the lactate level (Lac), glucose content (Glu), residual base (BEecf), pH, bicarbonate (HCO3-), high iron haemoglobin (MetHb). White blood cells (WBC), platelets (PLT), haemoglobin content (Hb), carboxyhaemoglobin (COHb) were detected using a quintuple classification. Blood creatinine (SCr), uric acid (UA), kidney-related indexes were detected using biochemistry instrument. Kidney tissues of the rats were taken after 24 h of resuscitation and after execution, and the inflammation of kidneys of the rats of the two groups was analyzed using HE staining. Fluorescence staining was used to detect the level of ROS in the kidneys of rats in both groups. [Results] At RS 0h, the Beecf, Glu and Lac levels of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group, and the pH level of rats in the SMI+HBOC-CHP01 group was significantly higher than that of rats in the HBOC-CHP01 group, and the Glu levels of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group at RS 1h. At RS 0h, the WBC, PLT and COHb contents of rats in the SMI+HBOC-CHP01 group were all significantly higher than those of rats in the HBOC-CHP01 group, and at RS 1h, the WBC content of rats in the SMI+HBOC-CHP01 group was significantly higher than that of rats in the HBOC-CHP01 group; at RS 1h, the UA content of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC-CHP01 group; at RS 24h, the SCr content of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC-CHP01 group; at RS 24h, the inflammation level of kidney tissues of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC -CHP01 group rats, and the ROS and MPO levels in the kidney tissues of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group. [Conclusion] The combination of Salvia miltiorrhiza injection during the resuscitation of rats with severe haemorrhagic shock by HBOC-CHP01 can alleviate renal injury by reducing inflammatory response and oxidative stress.

To summarize the nursing experience of a patient with anthrax meningoencephalitis caused by cerebrospinal fluid rhinorrhea post-trauma.Nursing points include strict implementation of isolation and hospital infection prevention and control measures;to standardize the use of antibiotics throughout the process to prevent and manage antimicrobial-related diarrhea;to pay attention to the nutritional status of patients and develop individualized nutritional support programs based on evidence;to implement refined airway care to prevent pulmonary complications;to strengthen disease observation,identify intracranial hemorrhage in time;to provide patient and family-centered psychological empowerment in the whole process.After 66 days of careful treatment and nursing care,the patient recovered and was discharged.A follow-up visit after 2 months of discharge showed a good recovery.

Objective To investigate the medication law of Professor Wang Xuefeng in the treatment of transient tic disorders in children using data mining techniques;To clarify clinical thoughts and academic experience in the treatment of this disease.Methods The outpatient medical records of Professor Wang Xuefeng treated children with transient tic disorder in Affiliated Hospital of Liaoning University of Traditional Chinese Medicine from September 2021 to September 2024 were collected,and the prescriptions for patients'first visit were input after screening and sorting.Excel 2019 was used to establish the database.SPSS Modeler 18.0 was used to analyze the association rules of high-frequency drugs.SPSS Statistics 27.0 was used for clustering analysis.Gephi 0.10.1 was used for complex network analysis,and medication law was summarized.Results A total of 223 TCM prescriptions meeting the standard were included,involving 116 kinds of Chinese materia medica,with a total frequency of 3 668,and 30 high-frequency drugs were screened.The properties were mainly cold,neutral,and warm and the tastes were mainly pungent,bitter and sweet.The meridians were mainly liver,lung and heart meridians.The efficacy was mainly for suppressing hyperactive liver for calming endogenous wind,relieving surface and clearing heat;the results of association rules analysis showed that the drug combination with the highest correlation was Paeoniae Radix alba-Glycyrrhizae Radix et Rhizoma Praeparata cum Melle-Acori Tatarinowii Rhizoma;the high-frequency drugs were grouped into 9 categories by clustering analysis;a core drug combination consisting of 10 kinds of highly correlated Chinese materia medica was obtained by complex network analysis.Conclusion Professor Wang Xuefeng believes that transient tic disorders is mainly in the lung and liver,the etiology is due to the wind and phlegm,the pathogenesis is the external pathogens invading the lung,the pathogenic qi is reluctant to leave,the five viscera are disharmonious,the producing phlegm and turbid,the internal and external connection,the wind phlegm consolidation,the flowing meridians,thus causing twitching.It is necessary to take into account the whole process of disease development,with clear medication,fewer tastes,refined prescription and special efficacy.

Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.

Objective:To analyze the efficacy of phantom-less quantitative computer tomography (PL-QCT) for the diagnosis of osteoporosis in patients with lumbar degenerative diseases.Methods:From October 2021 to October 2023, a total of 1 248 patients with lumbar degenerative disease who did not receive anti-osteoporosis treatment in the Department of Spine Surgery, Tianjin Hospital were retrospectively analyzed. There were 520 males and 728 females, aged 62.31±9.37 years (range, 40-87 years), height 1.66±0.08 m (range, 1.43-1.89 m), weight 69.04±8.27 kg (range, 49-93 kg). The mean body mass index was 26.11±3.67 kg/m 2 (range, 14.40-37.11 kg/m 2). Dual-energy X-ray absorptiometry (DXA) and PL-QCT were used to diagnose osteoporosis, and the detection rates of the two diagnostic methods were compared. The receiver operating characteristic (ROC) curve of PL-QCT for the diagnosis of osteoporosis was drawn, the area under the curve (AUC) and 95% confidence interval (CI), sensitivity and specificity were calculated. Results:Among 1 248 patients with lumbar degenerative diseases, 626 (50.2%) were diagnosed as osteoporosis by PL-QCT, 423(33.9%) were diagnosed by spine DXA, 488(39.1%) were diagnosed by hip DXA and 539 patients(43.2%) were diagnosed by dual-site DXA. The detection rate of osteoporosis of PL-QCT was higher than that of spine DXA (χ 2=193.557, P<0.001), hip DXA (χ 2=322.201, P<0.001) and dual-site DXA (χ 2=94.683, P<0.001), and the difference was statistically significant. Taking the diagnostic results of spinal DXA for osteoporosis as a reference, the ROC curve of PL-QCT for the diagnosis of osteoporosis showed a sensitivity of 79%, a specificity of 81%, and an AUC and 95% CI of 0.82(0.79, 0.85). Taking the diagnostic results of hip DXA for osteoporosis as a reference, the ROC curve of PL-QCT for the diagnosis of osteoporosis showed a sensitivity of 85%, a specificity of 55%, and an AUC and 95% CI of 0.75(0.71, 0.78). Taking the diagnostic results of two-site DXA for osteoporosis as a reference, the ROC curve of PL-QCT for the diagnosis of osteoporosis showed a sensitivity of 72%, a specificity of 75%, and an AUC and 95% CI of 0.81(0.78, 0.83). Conclusion:Compared with DXA, PL-QCT has a higher detection rate of osteoporosis in patients with degenerative lumbar spine disease and good diagnostic efficacy.

Objective:To investigate the clinical efficacy of lumbar lateral interbody fusion (LLIF) versus posterior lumbar interbody fusion (PLIF) in the treatment of severe lumbar spinal stenosis.Methods:The data of patients with severe lumbar spinal stenosis who underwent LLIF or PLIF from February 2019 to December 2023 were retrospectively analyzed. There were 30 patients in the LLIF group, 10 males and 20 females, aged 62.7±5.6 years (range, 53-74 years), including 21 cases of single segment and 9 cases of double segment. There were 46 patients in the PLIF group, including 20 males and 26 females, aged 63.2±8.4 years (range, 43-75 years), 40 cases of single segment and 6 cases of double segment. The visual analogue scale (VAS), Oswestry disability index (ODI), intervertebral space height, intervertebral foramen height and postoperative complications were compared between the two groups.Results:All patients were followed up for an average of 21.3±6.4 months (range, 12-32 months). The intraoperative blood loss in the LLIF group was 112.2±76.9 ml, which was significantly lower than 193.9±88.2 ml in the PLIF group ( P<0.05). The VAS scores of back pain and leg pain after operation were significantly lower than those before operation in the two groups ( P<0.05). There was no statistically significant difference between groups in back pain VAS scores at preoperative, 6 months postoperative, and final follow-up ( P>0.05); the back pain VAS score at 1 month postoperatively in the LLIF group was 1.6±1.2, which was less than 2.8±0.7 in the PLIF group ( P<0.05). There was no statistically significant difference between groups in leg pain VAS scores at preoperative, 1 month postoperative, and 6 months postoperative ( P>0.05); the leg pain VAS score at the final follow-up in the LLIF group was 1.2±1.5, which was smaller than 1.8±1.0 in the PLIF group ( P<0.05). The postoperative ODI was smaller than the preoperative one in both groups, and the difference was statistically significant ( P<0.05); the preoperative, 1-month postoperative, 6-month postoperative, and final follow-up ODIs in the LLIF group were 45.7%±16.0%, 17.9%±12.0%, 16.2%±11.6%, and 15.7%±11.7%, and those in the PLIF group were 47.9%±15.4%, 20.1%±9.3%, 16.9%±10.6%, and 14.6%±11.0% in the PLIF group, and the difference between the groups was not statistically significant ( P>0.05). The preoperative intervertebral space height in the LLIF group was 10.6±2.0 mm, which was smaller than that in the PLIF group 11.8±2.2 mm ( P<0.05). The intervertebral space heights in the immediate postoperative period and at the final follow-up were 13.3±2.3 mm and 12.3±2.2 mm in the LLIF group and 13.7±1.7 mm and 13.0±1.9 mm in the PLIF group ( P>0.05). The preoperative intervertebral foraminal height in the LLIF group was 18.0±3.2 mm, which was smaller than that of 19.7±2.4 mm in the PLIF group ( P<0.05); the intervertebral foraminal heights in the immediate postoperative period and at the final follow-up were 21.4±2.5 mm and 20.2±2.4 mm in the LLIF group, and in the PLIF group were 20.7±2.4 mm and 19.7±2.6 mm in the PLIF group ( P>0.05). In the LLIF group, 2 cases had femoral nerve injury and 2 cases had transient back pain after operation. There were 2 cases of cerebrospinal fluid leakage, 1 case of screw loosening, and 2 cases of deep vein thrombosis in the PLIF group. In the PLIF group, 2 patients underwent revision, including 1 case due to cage displacement and 1 case due to screw malposition. The fusion settling rate was 21% (8/39) in the LLIF group and 12% (6/52) in the PLIF group ( P>0.05). Conclusion:Both LLIF and PLIF can effectively restore the intervertebral height, improve the lumbar function and the symptoms of back and leg pain in the treatment of severe lumbar spinal stenosis.

Objective:To investigate the risk factors for complications following lateral lumbar interbody fusion (LLIF) surgery.Methods:A retrospective analysis was conducted on 196 patients who underwent LLIF surgery via the psoas major muscle approach in the Department of Spinal Surgery, Tianjin Hospital, Tianjin University, from October 2018 to July 2024. The age, gender, body mass index (BMI), presence of hypertension, type 2 diabetes, heart disease, osteoporosis, history of lumbar surgery, history of abdominal surgery, history of abdominal inflammation, smoking status, operative time, surgical segments (whether the surgery was single-segment or multi-segments), and use of internal fixation or not were compared between patients with and without postoperative complications, the indicators with P<0.10 were included in the binary variable logistic regression analysis, and determine the independent risk factors for complications after LLIF surgery. Complications included anterior thigh symptoms (pain, numbness, weakness), cage subsidence, surgery-related complications (nerve injury, surgical site infection, postoperative buttock pain, urinary and fecal incontinence, etc.), and medical complications (cerebrovascular accident, deep vein thrombosis, urinary tract infection, etc.). Results:All 196 patients were followed up for 27.02 (12.6, 40.69) months. Postoperative complications occurred in 71 cases (96 times), and no complications occurred in 125 cases. In the complication group, there were 15 males and 56 females with a mean age of 61.82±7.57 years; in the non-complication group, there were 43 males and 82 females with a mean age of 62.00± 8.39 years. In the complication group, there were 43 cases of anterior thigh symptoms, 38 cases of cage subsidence, 11 cases of surgical operation-related complications, and 4 cases of medical complications. There were statistically significant differences in gender (χ 2=3.829, P=0.051), operation time ( t=2.391, P=0.018), and surgical segment (χ 2=4.245, P=0.039) between the complication group and the non-complication group. No statistically significant differences were found in age, gender, BMI, presence of hypertension, type 2 diabetes, heart disease, osteoporosis, history of lumbar surgery, history of abdominal surgery, history of abdominal inflammation, smoking status, or use of internal fixation ( P>0.10). Binary variable logistic regression analysis indicated that prolonged operative time [ OR=1.007, 95% CI(1.001, 1.013), P=0.019] and multilevel surgery [ OR=2.099, 95% CI(1.095, 4.025), P=0.026] were independent risk factors for complications following LLIF. Conclusion:Prolonged operative time and multi-segments surgery are independent risk factors for complications following LLIF.

OBJECTIVE: To investigate the effectiveness of one-stage direct suture technique under arthroscopy for the treatment of anterior cruciate ligament (ACL) injury combined with anterior meniscus root injury. METHODS: The clinical data of 9 patients with ACL injury or tibial intercondylar eminence fracture combined with anterior meniscus root injury who were admitted between September 2017 and September 2024 and met the selection criteria were retrospectively analyzed. There were 3 males and 6 females, aged 21-62 years, with an average age of 37.1 years. The time from injury to surgery ranged from 5 days to 5 years, with a median time of 40 days. Among them, 5 cases had ACL injury, including 4 cases of type 1 and 1 case of type 2 according to modified Sherman classification; 4 cases had tibial intercondylar eminence fracture, including 3 cases of type 3 and 1 case of type 2 according to Meyers-McKeever classification. There were 7 cases of anterior root injury of lateral meniscus and 2 cases of anterior root injury of medial meniscus. The preoperative International Knee Documentation Committee (IKDC) score was 45.0±12.3, and Lysholm score was 49.2±12.4. Preoperatively, 7 cases were positive in anterior drawer test, Lachman test, and McMurray test, while 2 cases could not complete the test due to pain limitation. Preoperatively and at last follow-up, IKDC score and Lysholm score were used to evaluate knee joint function, anterior drawer test and Lachman test were used to evaluate knee joint stability, and McMurray test was used to evaluate meniscus condition. RESULTS: The operation time was 30-100 minutes, with an average of 64.2 minutes; the total hospital stay was 2-12 days, with an average of 4.5 days; the postoperative hospital stay was 1-4 days, with an average of 1.8 days. All incisions healed by first intention without surgery-related complications. All 9 patients were followed up 2-30 months, with an average of 18.8 months. No internal fixation-related complications occurred during follow-up. At last follow-up, MRI review showed good ligament tension, and CT showed good fracture healing. The results of anterior drawer test and Lachman test were all negative. McMurray test was negative in all cases. The IKDC score was 88.3±5.1, and Lysholm score was 88.3±5.6, both showing significant improvement compared to preoperative scores ( t=14.001, P<0.001; t=10.192, P<0.001). CONCLUSION One-stage direct suture technique under arthroscopy for repairing ACL injury or tibial intercondylar eminence fracture combined with anterior meniscus root injury can achieve good effectiveness without fixation device-related complications.
Humans ; Adult ; Male ; Female ; Arthroscopy/methods* ; Middle Aged ; Anterior Cruciate Ligament Injuries/surgery* ; Tibial Meniscus Injuries/surgery* ; Retrospective Studies ; Suture Techniques ; Young Adult ; Treatment Outcome ; Tibial Fractures/surgery* ; Anterior Cruciate Ligament Reconstruction/methods* ; Menisci, Tibial/surgery*

This article systematically analyzes the historical evolution of the name， origin， quality evaluation， harvesting， processing and other aspects of Picrorhizae Rhizoma by referring to the medical books， prescription books， and other documents of the past dynasties， combined with relevant modern research materials， in order to provide a basis for the development and utilization of famous classical formulas containing this medicinal herb. The research results indicate that Picrorhizae Rhizoma was first recorded in New Revised Materia Medica from the Tang dynasty. Throughout history， Huhuanglian has been used as its official name， and there are also aliases such as Gehu Luze， Jiahuanglian and Hulian. The main source of past dynasties is the the rhizomes of Picrorhiza kurrooa and P. scrophulariiflora. In ancient times， Picrorhizae Rhizoma was mainly imported by foreign traders via Guangzhou and other regions， and also produced in China， mainly in Xizang. In ancient times， it was harvested and dried in early August of the lunar calendar， while in modern times， it is mostly harvested from July to September， with the best quality being those with thick and crispy rhizomes without impurities， and bitter taste. Throughout history， Picrorhizae Rhizoma was collected， washed， sliced， and dried before being used as a raw material for medicine， it has a bitter and cold taste， mainly used to treat bone steaming， hot flashes， infantile chancre fever， and dysentery. There is no significant difference in taste and efficacy between ancient and modern times. Based on the research results， it is recommended that the rhizomes of P. scrophulariiflora in the 2020 edition of Chinese Pharmacopoeia， or the rhizomes of P. kurrooa， can be used in famous classical formulas containing this medicinal herb， which can be processed according to the processing requirements marked by the original formula. For those without clear processing requirements， the dried raw products are used as medicine.