Objective:To identify and validate secreted effector proteins of Chlamydia psittaci ( C. psittaci) through bioinformatic prediction and experimental verification, and to characterize their subcellular localization in host cells. Methods:Potential effector proteins were predicted using bioinformatics tools. Candidate effectors were fused to β-lactamase through the constructed expression vectors, and these vectors were transformed into C. psittaci. The secretion of these candidate effectors was evaluated by β-lactamase translocation assays. Eukaryotic expression vectors of confirmed effectors were transfected into host cells to determine their intracellular localization patterns. Results:Bioinformatic analysis identified 29 candidate effector proteins. Experimental validation confirmed the secretion of five effectors, with four exhibiting cytoplasmic localization and one displaying nuclear localization in host cells.Conclusion:This study characterizes five novel C. psittaci secreted effector proteins, providing critical insights for investigating the molecular pathogenesis of psittacosis.

Objective:To identify and validate secreted effector proteins of Chlamydia psittaci ( C. psittaci) through bioinformatic prediction and experimental verification, and to characterize their subcellular localization in host cells. Methods:Potential effector proteins were predicted using bioinformatics tools. Candidate effectors were fused to β-lactamase through the constructed expression vectors, and these vectors were transformed into C. psittaci. The secretion of these candidate effectors was evaluated by β-lactamase translocation assays. Eukaryotic expression vectors of confirmed effectors were transfected into host cells to determine their intracellular localization patterns. Results:Bioinformatic analysis identified 29 candidate effector proteins. Experimental validation confirmed the secretion of five effectors, with four exhibiting cytoplasmic localization and one displaying nuclear localization in host cells.Conclusion:This study characterizes five novel C. psittaci secreted effector proteins, providing critical insights for investigating the molecular pathogenesis of psittacosis.

Objective To investigate the prevalence of tick-borne rickettsial infections in selected areas of Liupanshui City, Guizhou Province, 2023, so as to provide insights into the management of tick-borne rickettsioses in the city. Methods Ticks were captured from the body surface of bovines and sheep in Gaoxing Village, Dashan Township, Liupanshui City, Guizhou Province during the period between April and June, 2023, and tick species were identified using morphological and molecular biological techniques. In addition, tick-borne Rickettsia was identified using a nested PCR assay, including spotted fever group rickettsiae (SFGR), Coxiella spp., Anaplasma spp., Ehrlichia spp., and Orientia spp., and positive amplified fragments were sequenced and aligned with known sequences accessed in the GenBank database. Results A total of 200 ticks were collected and all tick species were identified as Rhipicephalus microplus. Nestle PCR assay combined with sequencing identified ticks carrying Candidatus Rickettsia jingxinensis (40.50%), Coxiella burnetii (1.50%), and Coxiella-like endosymbionts (27.00%), and Anaplasma spp., Ehrlichia spp. or Orientsia spp. was not detected. Conclusions R. microplus carried Candidatus R. jingxinensis, C. burnetii, and Coxiella-like endosymbionts in selected areas of Liupanshui City, Guizhou Province. Intensified monitoring of tickborne rickettsial infections is needed in livestock and humans to reduce the damages caused by rickettsioses.

Objective A recombinant plasmid for CRISPR interference(CRISPRi)that incorporates dCas9 from Streptococcus pyogenes was constructed to investigate the function of Coxiella burnetii genes through a prompt,simple method.Methods A single guide RNA(sgRNA)sequence targeting C.burnetii dotB was integrated into a recombinant plasmid,which was then transformed into C.burnetii using electroporation.Next,dCas9 expression was induced with dehydrated tetracycline(aTC)to inhibit the transcription of dotB.Results The results demonstrated that dCas9 can be expressed normally in the CRISPRi system through aTC induction.Suppression of dotB was observed at the transcriptional and translational levels.In addition,the intracellular reproduction of C.burnetii significantly decreased after the suppression of dotB expression.Conclusion Silencing technology for the C.burnetii gene based on CRISPRi was established in this study,providing support for studying the biological functions of C.burnetii genes.

Objective:Investigate the safety and feasibility of using the new self-designed implanting applicator in vaginal three-dimensional intracavitary brachytherapy after hysterectomy for gynecological cancer, and to explore the clinical value of the self-designed implanting applicator.Methods:Sixty-two gynecological cancer patients who underwent brachytherapy in Sun Yat-sen University Cancer Center were selected in this study. Each patient received three-dimensional intracavitary brachytherapy because of the indication of postoperative radiotherapy. Each patient was treated with different types of self-designed implanting applicators according the condition of postoperative vagina,and the vaginal tube and implant needle were placed in the template according to the preset channnel. Based on the actual CT images, the high-risk clinical target volume (HR-CTV), and organs at risk were defined according to unified target area delineation criteria and then the brachytherapy plan was conducted. The prescription dose of high-risk clinical target volume (HR-CTV) was 5.5 Gy/time. The parameters such as target area, organs at risk volume and irradiated dose were evaluated by DVH diagram.Results:Sixty-two patients successfully completed brachytherapy under the guidance of self-designed implanting applicator. A total of 140 implantation treatments were performed. The total average dose of HR-CTV D90% was (575.48±22.30) cGy, the mean dose D 2cm3 of bladder, rectum and sigmoid colon were (328.69±102.71), (369.14±46.59) and (27.28±71.27) cGy, the small intestine did not drop the target area, so there was no statistics. There was statistical significance between target volume and organs at risk dose ( P<0.05). Conclusions:The new self-designed implanting applicator has obvious clinical advantages in vaginal three-dimensional intracavitary brachytherapy after hysterectomy for gynecological cancer, meets the requirements of the preset planning dose,and it is sample to operate and highly safe,which indicated a bright future of the clinical application.

Background Acute exposure to mercury chloride (HgCl₂) can cause liver damage. Whether oleanolic acid (OA) as a hepatoprotective drug can protect against liver injury induced by acute exposure to HgCl₂ and related mechanism of action remain unclear. Objective To investigate the protective effect and possible mechanism of OA on liver injury in mice caused by acute exposure to HgCl₂. Methods Forty SPF C57BL/6 male mice were randomly divided into four groups with 10 mice in each group according to body weight. The four groups were named control group, OA group (300 mg·kg⁻¹), HgCl₂ group (5 mg·kg⁻¹), and OA + HgCl₂ group (300 mg·kg⁻¹ OA + 5mg·kg⁻¹ Hgcl₂). Soybean oil and OA solution were administered intragastric once a day for two consecutive days. HgCl₂ solution was injected intraperitoneally 2 h after the second intragastric administration. Mice were sacrificed after 48 h, and their serum and liver were collected. Liver coefficient was calculated. The changes of liver structure and iron deposition were observed by hematoxylin-eosin (HE) staining and Prussian blue staining. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), total superoxide dismutase (T-SOD), reduced glutathione (GSH), malondialdehyde (MDA), and tissue iron content were measured with commercial kits. Western blotting was used to detect nuclear factor erythroid-2 related factor 2 (Nrf2), heme oxygenase 1 (HO-1), glutathione peroxidase 4 (Gpx4), transferrin receptor 1 (TFR1,) and solute carrier family 7 member 11 (SLC7A11). Results The AST and ALT levels of the HgCl₂ group were (76.447±9.695) U·g⁻¹ and (98.563±24.673)U·g⁻¹, respectively, which were higher than those of the control group (P<0.05). After the OA pretreatment, the liver coefficient and the above indexes were decreased to (4.769±0.237)%, (57.086±10.087) U·g⁻¹, and (87.294±27.181)U·g⁻¹, respectively. The liver coefficient and AST level of the OA + HgCl₂ group were significantly different from those of the HgCl₂ group (P<0.05). After acute exposure to HgCl₂, the hepatocytes of mice were disordered, accompanied by inflammatory infiltration, positive blue particles appeared in Prussian blue staining of liver tissue, and the above changes in liver tissue were alleviated after the OA pretreatment. The iron content in the HgCl₂ group was (3.646±0.238) μmol·g⁻¹, which was higher than that in the control group, (2.948±0.308) μmol·g⁻¹. After the OA pretreatment, the iron content decreased to (3.429±0.415) μmol·g⁻¹. Compared with the control group, acute exposure to HgCl₂ resulted in decreased levels of GSH and T-SOD, decreased protein expression levels of Nrf2, HO-1, SLC7A11, and Gpx4, increased level of MDA, and increased protein expression level of TFR1 (P<0.05). After the OA pretreatment, all indicators were improved including increased GSH level, decreased MDA level, increased Nrf2, HO-1, and SLC7A11 protein expression levels, and decreased TFR1 protein expression level; compared with the HgCl₂ group, the differences were statistically significant (P<0.05). Conclusion Acute HgCl₂ exposure could induce liver injury in mice, and its mechanism may involve iron overload and ferroptosis. OA may alleviate the liver injury caused by acute HgCl₂ exposure by affecting iron overload and the ferroptosis-related protein expression.

Objective : To investigation of the interaction between interstitial cells of Cajal (ICCs) and connexin 43 ( Cx43) in bladder contraction and its significance． Methods : Eighty male guinea pigs were randomly divided into blank control group，Glivec group，Gap27 group and Glivec + Gap27 group．Four groups of guinea pigs were per- fused with saline，Glivec，Gap 27，and Glivec + Gap 27 every morning for 2 months．Success of urodynamic testing model after 2 months．Bladder tissue was collected for an in vitro muscle strip test to detect muscle contraction in each group．Correlation between c-Kit and Cx43 was detected by immunofluorescence．The interaction between c- Kit and Cx43 in the bladder was further validated by qRT-PCR and Western blot．Ultrastructural changes in the muscle layer of the bladder were observed by electron microscopy. Results : Urodynamics revealed increased blad- der compliance in the experimental group compared to the blank control group (P＜0. 05) ; bladder compliance increased in the Glivec + Gap27 group compared to the Glivec and Gap27 groups (P＜0. 01) ．In vitro muscle strip experiments revealed that the frequency and tone of bladder muscle strip contractions were lower in the experimental group compared to the blank control group (P＜0. 05) ，and that muscle strip contractions were weaker in the ex- perimental group after administration of acetylcholine (ACH) compared to the control group(P＜0. 05) ．Immuno- fluorescence showed that c-Kit was co-expressed with Cx43 on ICCs cells．qRT-PCR and Western blot suggested that the protein expression level and gene expression level of Cx43 in bladder tissues were lower after inhibition of c-Kit than in the blank control (P＜0. 05) ; after inhibition of Cx43，the protein expression level and gene expres- sion level of c-Kit in bladder tissues the levels of c-Kit protein expression and gene expression in bladder tissues were lower than those in the blank control group (P＜0. 05) ．Electron microscopy revealed that the mitochondrial structure of bladder smooth muscle was disrupted after simultaneous inhibition of c-Kit and Cx43． Conclusion Cx43 is expressed on bladder ICCs and the two may be jointly involved in regulating bladder contractile function ; the joint reduction of Cx43 and c-Kit may have disrupted the mitochondria of bladder smooth muscle，affecting its function and consequently bladder contractile function.

Objective : To investigate the effect of the combination of stem cell leukemia(SCL)recombinant lentivirus and connexin 43 ( Cx43 ) recombinant lentivirus on the function of diabetic cystipathy ( DCP) in guinea pigs. Methods : After 90 healthy guinea pigs were fed normally for 1 week, streptozotocin(STZ)was injected intraperito⁃ neally at 200 mg/kg in a single dose, and random blood glucose was monitored weekly. After 12 weeks of normal feeding, 40 guinea pigs meeting the criteria were screened by urodynamic examination and randomly divided into 4 groups: diabetic group, SCL group, Cx43 group, and SCL + Cx43 group. In the diabetic group, 0. 2 ml of empty lentivirus without gene was instilled into the bladder through the urethra, in the SCL and Cx43 groups, 0. 2 ml of SCL lentivirus and Cx43 lentivirus were instilled using the same method, in the SCL + Cx43 lentivirus group, 0. 2 ml of each SCL and Cx43 lentivirus was instilled through the urethra. After 14 days of transfection, urodynamic examination was performed, and then the guinea pigs were executed after the examination, and the bladders were quickly removed for frozen bladder sections and fluorescent double staining. Results : There were no differences in urodynamic examinations between the SCL and Cx43 groups compared to the diabetic group( P > 0. 05 ) . Urodynamic examination in the SCL + Cx43 group showed improvement in detrusor pressure, abdominal pressure and bladder pressure compared to the diabetic group(P < 0. 05) . In laser confocal experiments, the number of interstitial cells of Cajal (ICC) was reduced in the diabetic group, and the spindle structure and cell protrusions were obviously destroyed and appeared in a state of cell lysis. In the SCL and Cx43 groups, there was an improvement in the spindle structure and cell protrusion of ICC⁃like cells, and there was no significant change in the number of cells. In the SCL + Cx43 group, there was an increase in the number of ICC⁃like cells, a significant improvement in the spindle structure and cell protrusion, and the formation of ICC⁃dimers. Conclusion Transurethral co⁃infusion of SCL and Cx43 gene recombinant lentivirus can be successfully transfected in guinea pig DCP bladder, which can restore the number and structure of damaged ICC cells and form ICC dimer structure, improve the pressure of diabetic bladder forced urinary muscle, improve the weakness of urination, ventral pressure voiding and other characteristics. It provides a new direction for the treatment of DCP.

Objective:To investigate the clinical feasibility of the Unity radiotherapy system guided by magnetic resonance imaging.Methods:Twenty-four patients were enrolled and received a total of 384 fractions of treatment at Unity system. According to the treatment site, all patients were divided into head-neck, abdomen-thorax, pelvic, spine and limb groups. The patients were set-up without external laser. And then, the time required at different stages in online treatment process and the registration error of each fraction were separately calculated. The geometric deformations of MR images were weekly measured by using MR geometric deformation phantom. At last, the Arccheck was used to perform the dose verification of reference plan, online plan and offline plan.Results:The mean duration of radiotherapy in the five groups were 29.1, 27.6, 26.6, 25.6 and 32.0 min, respectively. The set-up errors in the left-right, superior-inferior and anterior-posterior direction in the five groups were: head-neck group (0.08±0.06 cm, 0.16±0.13 cm, 0.08±0.05 cm), abdomen-thorax group (0.23±0.18 cm, 0.50±0.47 cm, 0.12±0.1 cm), pelvic group (0.25±0.19 cm, 0.32±0.25 cm, 0.11±0.09 cm), spine group (0.46±0.38 cm, 0.26±0.26 cm, 0.13±0.07 cm) and limb group (0.33±0.30 cm, 0.34±0.23 cm, 0.08±0.06 cm), respectively. In the central region, the geometric deformation of MR was less than 0.3 mm, and that of the sphere with a diameter of 500 mm was less than 2.1 mm. The meanγ pass rate of the reference plan, online plan and offline plan were 97.92%, 97.84% and 94.58%, respectively.Conclusions:MR-guided radiotherapy has great potential for clinical application, whereas the process of Unity system is relatively complex. The synergy of different departments has a great impact on the treatment, which needs further optimization.

Pulmonary hypertension (PH) is a severe pathophysiological condition characterized by pulmonary artery remodeling and continuous increases in pulmonary artery pressure, which may eventually develop to right heart failure and death. Although newly discovered and incredible treatment strategies in recent years have improved the prognosis of PH, limited types of effective and economical drugs for PH still makes it as a life-threatening disease. Some drugs from Chinese materia medica (CMM) have been traditionally applied in the treatment of lung diseases. Accumulating evidence suggests active pharmaceutical ingredients (APIs) derived from those medicines brings promising future for the prevention and treatment of PH. In this review, we summarized the pharmacological effects of APIs derived from CMM which are potent in treating PH, so as to provide new thoughts for initial drug discovery and identification of potential therapeutic strategies in alternative medicine for PH.
China ; Drugs, Chinese Herbal ; Humans ; Hypertension, Pulmonary/drug therapy* ; Materia Medica ; Medicine, Chinese Traditional