Objective To explore the application value of metagenomic next-generation sequen-cing(mNGS)technology in the diagnosis of spinal tuberculosis.Methods A total of 129 patients with suspected spinal tuberculosis admitted from January 2021 to January 2023 were selected as study subjects.Lesion tissue samples were collected intraoperatively and subjected to conventional microbio-logical testing(CMT),Mycobacterium tuberculosis DNA(MTB-DNA)amplification testing,and mNGS testing.The diagnostic efficacy of different testing methods was compared using results of com-prehensive clinical diagnosis as the gold standard.Results Among 129 patients,101(78.29％)were confirmed to have spinal tuberculosis,and 28(21.71％)had other spinal infections.Using clinical results as the diagnostic gold standard,the sensitivity of mNGS was 94.06％(95/101),and specificity was 89.29％(25/28);the sensitivity of MTB-DNA amplification was 90.10％(91/101),and specificity was 89.29％(25/28);the sensitivity of CMT was 86.14％(87/101),and specifici-ty was 85.71％(24/28).Compared with MTB-DNA amplification and CMT,mNGS showed the highest consistency with clinical results,and its consistency in detecting different lesion sites was also optimal,with statistically significant differences(P＜0.05).Conclusion mNGS testing has high diagnostic value for spinal tuberculosis and can provide a reference for clinical diagnosis,thereby guiding clinical decision-making.

Objective To explore the efficacy and safety of platelet-derived growth factor(PDGF)combined with allograft bone transplantation in the treatment of spinal tuberculosis.Methods A total of 177 patients with lumbar tuberculosis admitted to the 4th People's Hospital of Qinghai Province from August 2018 to August 2023 were selected as the research subjects.Patients were divided into control group(n=49)and observation group(n=128)based on the source of the transplanted bone.All patients underwent at least 2 weeks of standard quadruple anti-tuberculosis chemotherapy before surgery.Patients in the control group received PDGF combined with autograft bone transplantation,while patients in the observation group received PDGF combined with allograft bone transplantation.The surgical duration,intraoperative blood loss,and length of hospital stay of patients in the two groups were recorded;the erythrocyte sedimentation rate(ESR)and serum C-reactive protein(CRP)levels of patients in the two groups were compared before surgery and at 1,3,6 months after surgery.Preoperative CT and magnetic resonance imaging(MRI)examinations were performed,and postoperative CT and MRI were performed after bone fusion was completed to compare the changes in Cobb angle before and after surgery.The visual analogue scale(VAS)was used to assess the pain degree in the lumbar region before surgery and at 1,3,6 months after surgery.The VAS scores of patients in the two groups,VAS scores of male patients in the two groups,and VAS scores of female patients in the two groups were compared before and after surgery,respectively.Results There was no statistically significant difference in surgical duration and length of hospital stay between the observation group and the control group(P＞0.05).The intraoperative blood loss of patients in the observation group was significantly less than that in the control group(P＜0.05).There was no statistically significant difference in Cobb angle before and after surgery between the two groups(P＞0.05).The postoperative Cobb angle significantly decreased in both groups when compared to preoperative values(P＜0.05).The VAS scores of patients in both groups decreased sequentially before surgery and at 1,3,6 months after surgery,with statistically significant differences in intra-group pairwise comparisons(P＜0.05).The VAS scores of male patients in both groups decreased sequentially before surgery and at 1,3,6 months after surgery,with statistically significant differences in intra-group pairwise comparisons(P＜0.05).The VAS scores of female patients in both groups also decreased sequentially before surgery and at 1,3,6 months after surgery,with statistically significant differences in intra-group pairwise comparisons(P＜0.05).There was no statistically significant difference in VAS scores between the observation group and the control group before surgery and at 1,6 months after surgery(P＞0.05);the VAS score of patients in the observation group was significantly lower than that in the control group at 3 months after surgery(P＜0.05).There was no statistically significant difference in VAS scores between male patients in the observation group and male patients in the control group before surgery and at 1,3,6 months after surgery(P＞0.05);the VAS score of male patients in the observation group was significantly lower than that in the control group at 3 months after surgery(P＜0.05).There was no statistically significant difference in VAS scores between female patients in the observation group and female patients in the control group before surgery and at 1,6 months after surgery(P＞0.05);the VAS score of female patients in the observation group was significantly lower than that in the control group at 3 months after surgery(P＜0.05).The ESR of patients in both groups decreased sequentially before surgery and at 1,3,6 months after surgery,with statistically significant differences in intra-group pairwise comparisons(P＜0.05).The serum CRP levels of patients in both groups also decreased sequentially before surgery and at 1,3,6 months after surgery,with statistically significant differences in intra-group pairwise comparisons(P＜0.05).There was no statistically significant difference in ESR between the observation group and the control group before surgery and at 1,3,6 months after surgery(P＞0.05).There was no statistically significant difference in serum CRP level between the observation group and the control group before surgery and at 1,6 months after surgery(P＞0.05);the serum CRP level of patients in the observation group was significantly higher than that in the control group at 3 months after surgery(P＜0.05).Conclusion The effect of PDGF combined with allograft bone transplantation in the treatment of spinal tuberculosis is comparable to that of autograft bone transplantation,but PDGF combined with allograft bone transplantation can significantly reduce postoperative pain degree,improve patient comfort,avoid additional damage caused by autograft bone transplantation,and reduce the physical burden on patients.It can be considered a safe and reliable surgical method for bone grafting in lumbar tuberculosis surgery.

Objective:To analyze the value of high-risk human papillomavirus (HR-HPV) persistent infection in the follow-up of expectant treatment for cervical intraepithelial neoplasia (CIN) Ⅱ in patients aged ≤40 years.Methods:A prospective cohort study was conducted. Women aged ≤40 years with fertility needs, cervical transformation zone type Ⅰ-Ⅱ, and CIN Ⅱ pathology under colposcopy-guided biopsy were selected from the cervical outpatient clinic of Shenzhen Maternity and Child Healthcare Hospital, Southern Medical University. HR-HPV genotyping test, cytological examination, and colposcopy evaluation were used for regular follow-up for 2 years. The natural outcomes of the lesions were obesrved, and the predictive value of HR-HPV persistent infection in the persistence or progression of CIN Ⅱ lesions was analyzed.Results:A total of 135 eligible patients were collected, and completed the follow-up for (26±11) months. Of them, 88 patients (65.2%, 88/135) showed regression, 27 (20.0%, 27/135) persistence to CIN Ⅱ, and 20 (14.8%, 20/135) progression to CIN Ⅲ (no one cervical cancer). Among 135 patients with CIN Ⅱ, there were 68 cases (50.4%, 68/135) of HR-HPV persistent infection and 67 cases (49.6%, 67/135) of HR-HPV non-persistent infection, respectively. The persistence or progression rate of CINⅡ lesions in patients with HR-HPV persistent infection was significantly higher than that in patients with HR-HPV non-persistent infection [66.2% (45/68) vs 3.0% (2/67); χ2=59.38, P<0.001]. HR-HPV persistent infection was an independent risk factor for persistence or progression of CIN Ⅱlesions ( OR=30.93, 95% CI: 5.63-169.74, P<0.001); with predictive sensitivity and specificity of 95.7% (45/47) and 73.9% (65/88), respectively, the sensitivity and specificity of predicting the progression of CIN Ⅱ lesions were 100.0% (20/20) and 58.3% (67/115), respectively. Within the first year of follow-up, 86.4% (76/88) of CIN Ⅱ patients showed lesion regression, and 83.6% (56/67) of CIN Ⅱ patients experienced HR-HPV conversion to negative. The persistence or progression rate of CIN Ⅱ lesions in 42 patients with HPV 16 and (or) 18 (HPV 16/18) infection (47.6%, 20/42) at the time of enrollment was significantly higher than that in 93 patients with other HR-HPV (not HPV 16/18) infection (29.0%, 27/93; χ2=4.40, P=0.036); the progression rate of CIN Ⅱ in patients with HPV 16/18 persistent infection (44.4%, 12/27) was significantly higher than that in patients with other HR-HPV (not HPV 16/18) persistent infection (19.5%, 8/41; χ2=4.87, P=0.027). Conclusions:HR-HPV persistent infection is a sensitive indicator to predict persistence or progression of CIN Ⅱ lesions in patients aged ≤40 years, which could be used as the basis for risk stratification management of expectant treatment of CIN Ⅱ. For CIN Ⅱ lesion regression and HR-HPV negative conversion usually occur within the first year, the follow-up time could be shortened to 1 year to reduce the risk of disease progression; surgical treatment is recommended for patients with HR-HPV persistent infection, especially HPV 16/18 infection.

Objective:To analyze the value of high-risk human papillomavirus (HR-HPV) persistent infection in the follow-up of expectant treatment for cervical intraepithelial neoplasia (CIN) Ⅱ in patients aged ≤40 years.Methods:A prospective cohort study was conducted. Women aged ≤40 years with fertility needs, cervical transformation zone type Ⅰ-Ⅱ, and CIN Ⅱ pathology under colposcopy-guided biopsy were selected from the cervical outpatient clinic of Shenzhen Maternity and Child Healthcare Hospital, Southern Medical University. HR-HPV genotyping test, cytological examination, and colposcopy evaluation were used for regular follow-up for 2 years. The natural outcomes of the lesions were obesrved, and the predictive value of HR-HPV persistent infection in the persistence or progression of CIN Ⅱ lesions was analyzed.Results:A total of 135 eligible patients were collected, and completed the follow-up for (26±11) months. Of them, 88 patients (65.2%, 88/135) showed regression, 27 (20.0%, 27/135) persistence to CIN Ⅱ, and 20 (14.8%, 20/135) progression to CIN Ⅲ (no one cervical cancer). Among 135 patients with CIN Ⅱ, there were 68 cases (50.4%, 68/135) of HR-HPV persistent infection and 67 cases (49.6%, 67/135) of HR-HPV non-persistent infection, respectively. The persistence or progression rate of CINⅡ lesions in patients with HR-HPV persistent infection was significantly higher than that in patients with HR-HPV non-persistent infection [66.2% (45/68) vs 3.0% (2/67); χ2=59.38, P<0.001]. HR-HPV persistent infection was an independent risk factor for persistence or progression of CIN Ⅱlesions ( OR=30.93, 95% CI: 5.63-169.74, P<0.001); with predictive sensitivity and specificity of 95.7% (45/47) and 73.9% (65/88), respectively, the sensitivity and specificity of predicting the progression of CIN Ⅱ lesions were 100.0% (20/20) and 58.3% (67/115), respectively. Within the first year of follow-up, 86.4% (76/88) of CIN Ⅱ patients showed lesion regression, and 83.6% (56/67) of CIN Ⅱ patients experienced HR-HPV conversion to negative. The persistence or progression rate of CIN Ⅱ lesions in 42 patients with HPV 16 and (or) 18 (HPV 16/18) infection (47.6%, 20/42) at the time of enrollment was significantly higher than that in 93 patients with other HR-HPV (not HPV 16/18) infection (29.0%, 27/93; χ2=4.40, P=0.036); the progression rate of CIN Ⅱ in patients with HPV 16/18 persistent infection (44.4%, 12/27) was significantly higher than that in patients with other HR-HPV (not HPV 16/18) persistent infection (19.5%, 8/41; χ2=4.87, P=0.027). Conclusions:HR-HPV persistent infection is a sensitive indicator to predict persistence or progression of CIN Ⅱ lesions in patients aged ≤40 years, which could be used as the basis for risk stratification management of expectant treatment of CIN Ⅱ. For CIN Ⅱ lesion regression and HR-HPV negative conversion usually occur within the first year, the follow-up time could be shortened to 1 year to reduce the risk of disease progression; surgical treatment is recommended for patients with HR-HPV persistent infection, especially HPV 16/18 infection.

Background Plateau environment may pose a serious impact on the physiological and psychological stress of people stationed on a plateau, especially for those engaged in military training and occupational activities. There is an urgent need to find drugs to prevent and treat injuries caused by high-altitude environment. Objective To analyze the current status, hotspots, and future trends of domestic and international research in the field of prevention and treatment of high-altitude disease (HAD) by traditional Chinese medicine (TCM), and provide references for scientific research. Methods Web of Science Core Collection (WOSCC) and China National Knowledge Infrastructure (CNKI) were searched for literature on TCM and HAD published from inception to 2022. Excel, CiteSpace, VOSviewer, and RStudio softwares were used to conduct visual analysis on the number of publications, types of publications, journals, authors, research institutions, and keywords. Results A total of 501 publications were evaluated in the present study, including 443 Chinese publications and 58 English publications. The annual number of publications showed a rising trend. MA Huiping was the leading author in number of publications in Chinese (37 publications), and ZHANG Yi and MENG Xianli were the leading authors in the number of publications in English (both 8 publications), respectively. The institutions with the most publications in Chinese were The 940th Hospital of Joint Logistics Support Force of Chinese People's Liberation Army and Lanzhou General Hospital of Lanzhou Military Region (both 32 publications), and the institution with the most publications in English was Chengdu University of Traditional Chinese Medicine (8 publications), respectively. The Chinese and English journals with the largest number of publications were the Journal of High Altitude Medicine (39 publications) and the Journal of Ethnopharmacology (10 publications), respectively. The most highly cited Chinese and English literature included Effects of rhodiola on the free radical metabolism and serum creatine kinase after exercise at plateau (61 citations) and Anti-hypoxic activity at simulated high altitude was isolated in petroleum ether extract of Saussurea involucrate (68 citations) , respectively. The most frequent keywords in the Chinese and English literature were high altitude polycythemia and oxidative stress, respectively. The keyword time zone and emergence maps showed that the research hotspots in this field shifted from prevention and treatment of HAD to animal experiments, and then to mechanisms of action, in which oxidative stress, hypoxic injury, inflammation, and apoptosis were the main focuses. Conclusion The research of TCM against HAD is identified from early clinical observation to associations between clinical outcome variation and pharmacological mechanisms, and further to applying multi-omics techniques to explore the physical basis of TCM efficacy and mechanisms of action with focuses like TCM formula and single herb active ingredients, so as to elaborate potential scientific connotation of TCM against HAD.

Abstract: To investigate the differential expression of lncRNA SH3BP5-AS1 in hepatocellular carcinoma and the possible mechanism of promoting hepatocarcinogenesis. Methods: The expression of SH3BP5-AS1 in hepatocellular carcinoma tissue/paraneoplastic tissue(n=91) was detected by qRT-PCR. TCGA database visualization software UALCAN analyzed SH3BP5-AS1 expression in a large sample of hepatocellular carcinoma tissues(n= 371). The expression of SH3BP5-AS1 was detected by qRT-PCR in normal hepatocytes THLE-2 and hepatoma cell lines Huh 7, BEL-7405, SNU-387, Hep 3 B. After silencing SH3BP5-AS1 expression by siRNA technology, the effects of silencing SH3BP5-AS1 on the proliferation of Hep 3 B cells were detected by CCK-8 assay, cell clone formation assay and Edu assay, and the effects of silencing SH3BP5-AS1 on apoptosis and cycle of Hep 3 B cells were detected by flow-through assay. Based on the expression of SH3BP5-AS1 and TM6 SF2 in hepatocellular carcinoma tissues, the expression correlation was counted and analyzed. Results: SH3BP5-AS1 was highly expressed in hepatocellular carcinoma tissues. SH3BP5-AS1 was highly expressed in a large sample of hepatocellular carcinoma tissues from the TCGA database SH3BP5-AS1 was highly expressed in different hepatocellular carcinoma cell lines Huh 7, BEL-7405, SNU-387 and Hep 3 B to a different extent. Silencing of SH3BP5-AS1 by siRNA inhibited the proliferation ability of Hep 3 B cells, promoted apoptosis ability, affected the cell cycle and increased the number of G1 phase cells. SH3BP5-AS1 and TM6 SF2 expression were negatively correlated in hepatocellular carcinoma tissues. Conclusion SH3BP5-AS1 is an important oncogenic non-coding RNA that can provide a theoretical basis for tumour-targeted therapy in patients with hepatocellular carcinoma.

Objective:To synthesize Gd labeled probe targeting transporter protein(TSPO) 2-(8-amino-2-(4-chlorophenyl)imidazo[1, 2-a]pyridine-3-yl)- N, N-dipropylacetamide (CB86)-diethylene triamine pentaacetic acid (DTPA), and investigate its MRI effect in rheumatoid arthritis (RA) model. Methods:CB86-DTPA was prepared by coupling a bifunctional chelating agent, and then chelated with Gd to obtain MRI targeted contrast agent CB86-DTPA-Gd. The cytotoxicity, MR relaxation rate and in vitro stability of CB86-DTPA-Gd were determined. RA model was established with Freund′s adjuvant and the biodistribution study and MRI was performed. The RA lesion and its surrounding normal tissue were used as regions of interest (ROI) to calculate the signal to noise ratio (SNR). Independent-sample t test was used to analyze the data. Results:CB86-DTPA-Gd had excellent biosafety and a good MR relaxation rate ( r1=11.05 mmol·L -1·s -1). The survival rate of RAW264.7 cells and 4T1 cells was still more than 90% at the maximum concentration (20 μmol/L) of Gd 3+. CB86-DTPA-Gd also exhibited good stability in human serum and phosphate buffer saline solution (PBS; pH=7.4). The in vivo biodistribution showed that CB86-DTPA-Gd had better inflammatory targeting efficiency, and the uptake of Gd in the inflamed site of the ankle joint was still (2.33±0.29) percent dose rate per gram of tissue (%ID/g) at 120 min after injection. MicroMRI showed that the inflammation of the right ankle joint displayed significant enhancement after the injection of CB86-DTPA-Gd and Gd-DTPA. The SNR of CB86-DTPA-Gd group was up to 23.21±1.44, and the maximum intensification time was 90 min after injection, and can be significantly inhibited by CB86-DTPA at all time points ( t values: 6.083-12.451, all P<0.05), while the Gd-DTPA group had a strengthening time of 30 min after injection with the SNR of 16.12±1.24. Conclusion:CB86-DTPA-Gd shows good macrophage targeting and good uptake in arthritic reaction sites, and is expected to be a novel MRI molecular probe for peripheral inflammation imaging.

A viable spermatozoon is a prerequisite for fertilization in intracytoplasmic sperm injection (ICSI). Thus, it is crucial to select viable but immotile spermatozoa on the day of ICSI. We report conflicting results in the identification of viable but immotile spermatozoa between the eosin-nigrosin staining and the laser test, which resulted in confusion for embryologists during assisted reproductive technology (ART). Three patients’ semen samples that showed no motile spermatozoa are described in this report. To identify viable spermatozoa, we used both the eosin-nigrosin test and the laser test for each sample, and repeated the semen analysis twice in each patient. Viable but immotile spermatozoa selected by the laser test were used for ICSI. Viable spermatozoa were detected by both the eosin-nigrosin and laser tests in two patients (case 1, 95.00% vs. 24.21% and 92.68% vs. 22.22%; case 2, 41.18% vs. 23.48% and 39.81% vs. 22.52%), indicating consistent results between the two methods. In the third patient, the eosin-nigrosin test yielded viability rates of 20.75% and 19.14%, while the result of the laser test was 0%. Thus, testicular aspiration was performed to collect viable sperm from this patient. Normal fertilization was achieved after the injection of viable but immotile spermatozoa selected from these patients by the laser test, resulting in the birth of two healthy babies. Our study documents a case where the eosin-nigrosin test showed a limitation in identifying viable but immotile spermatozoa for ART, while the laser test may overcome this limitation. Larger samples may be required to corroborate the clinical value of the laser test.
Fertilization ; Humans ; Parturition ; Reproductive Techniques, Assisted ; Semen ; Semen Analysis ; Sperm Injections, Intracytoplasmic ; Spermatozoa

Objective To synthesize a novel 18 F labeled probe targeting translocator protein ( TSPO) ligand 2-( 5, 7-diethyl-2-( 4-( 2-fluoroethoxy ) phenyl ) pyrazolo [ 1, 5-a ] pyrimidin-3-yl )-N, N-diethylacet-amide (VUIIS1008), and evaluate its biodistribution and imaging in rheumatoid arthritis (RA) model. Methods The tosylate substrate was labeled with 18 F using a tosyloxy for fluorine nucleophilic aliphatic substitution to obtain 18 F-VUIIS1008. The labeling efficiency, radiochemical purity, and stability in vitro were determined. In vitro cellular uptake and competitive binding assay were performed on RAW264.7 mac-rophage cells. Biodistribution and microPET/CT imaging were investigated on RA mice established by Com-plete Freund's Adjuvant. Two-sample t test was used to analyze the data. Results 18 F-VUIIS1008 was syn-thesized with the labeling yield up to (41.00±5.00)％, the radiochemical purity>98.00％, and the specific radioactivity >1. 52 × 108 MBq/mmol. 18 F-VUIIS1008 was highly stable with the radiochemical purity >98. 00％ at 4 h after incubation in mouse serum. In vitro, it also exhibited high specific TSPO binding in RAW264.7 macrophage cells. The uptake ratio was (14.00±0.30)％ at 1 h after incubation, and decreased significantly ((4.00±0.70)％;t=12.894, P<0.05) after adding excessive unlabeled VUIIS1008. The half maximal inhibitory concentration (IC50) of 18F-VUIIS1008 binding to TSPO was 0.05 nmol/L in RAW264.7 macrophage cells. In vivo distribution results showed that the uptake of 18 F-VUIIS1008 in the left arthritic ankles reached the peak value of (1.33±0.02) percentage activity of injection dose per gram of tissue (％ID/g) at 1 h after injection. The radioactivity ratio of left ankle arthritic tissue to blood ( A/B) and to normal muscle ( A/M) was 4.40±0.22 and 1.65±0.07 respectively. MicroPET/CT imaging demonstrated that 18F-VUIIS1008 could specifically target and retained in the inflammation site. Conclusion 18 F-VUIIS1008 can be easily synthe-sized with high radiochemical purity and can clearly visualized in RA imaging with low background, suggesting its potential as a novel promising molecular probe targeting TSPO for RA PET imaging.

OBJECTIVE To study the cardiotoxicity of ophiopogonin D′(OPD′) for rat H9c2 cardio? myocytes. METHODS H9c2 cells were exposed to OPD′ 0.1, 1, 5, 10, 20, 25 and 50 μmol·L-1 for 24 h. Cell viability was examined by MTS assay, and the morphological changes in H9c2 cells were quanti? fied. The cell nucleus injury was examined by high content immune fluorescence screening and the morphological changes were observed under a fluorescence microscope. After treatment with OPD′ 0.1, 1, 5 and 10 μmol·L- 1 for 24 h, the effect on reactive oxygen species (ROS), mitochondrial mem? brane potential(MMP) and apoptosis was detected by flow cytometry. RESULTS The viability was sig? nificantly reduced following exposure to OPD′ 0.1, 1, 5, 10, 20, 25 and 50 μmol·L- 1 (P<0.05,P<0.01). The IC50 value was 9.9 μmol ·L- 1 and cell shrinkage and apoptosis occurred. The levels of ROS and apoptosis rate of H9c2 cells were significantly increased after exposure to OPD′ 0.1, 1, 5 and 10 μmol·L-1 for 24 h (P<0.05,P<0.01) and MMP markedly declined (P<0.05,P<0.01). CONCLUSION OPD′ has significent cytotoxicity on H9c2 cells. It may be related to inducing apopotsis pathways.