1.A reporter gene assays for bioactivity determination of human chorinonic gonadotropin
Ying HUANG ; Xiao-ming ZHANG ; He-yang LI ; Lü-yin WANG ; Hui ZHANG ; Ping LÜ ; Jing LI ; Xiang-dong GAO ; Cheng-gang LIANG
Acta Pharmaceutica Sinica 2024;59(2):432-438
		                        		
		                        			
		                        			 This study constructed a LHCGR-CRE-luc-HEK293 transgenic cell line according to the activation of the cAMP signaling pathway after recombinant human chorionic gonadotropin binding to the receptor. The biological activity of recombinant human chorionic gonadotropin was assayed using a luciferase assay system. The relative potency of the samples was calculated using four-parameter model. And the method conditions were optimized to validate the specificity, relative accuracy, precision and linearity of the method. The results showed that there was a quantitative potency relationship of human chorinonic gonadotropin (hCG) in the method and it was in accordance with the four-parameter curve. After optimization, the conditions were determined as hCG dilution concentration of 2.5 μg·mL-1, dilution ratio of 1∶4, cell number of 10 000-15 000 cells/well, and induction time of 6 h. The method had good specificity, relative accuracy with relative bias ranging from -8.9% to 3.4%, linear regression equation correlation coefficient of 0.996, intermediate precision geometric coefficient of variation ranging from 3.3% to 15.0%, and linearity range of 50% to 200%. This study successfully established and validated a reporter gene method to detect hCG biological activity, which can be used for hCG biological activity assay and quality control. 
		                        		
		                        		
		                        		
		                        	
2.Early gait analysis after total knee arthroplasty based on artificial intelligence dynamic image recognition
Ming ZHANG ; Ya-Nan SUI ; Cheng WANG ; Hao-Chong ZHANG ; Zhi-Wei CAI ; Quan-Lei ZHANG ; Yu ZHANG ; Tian-Tian XIA ; Xiao-Ran ZU ; Yi-Jian HUANG ; Cong-Shu HUANG ; Xiang LI
China Journal of Orthopaedics and Traumatology 2024;37(9):855-861
		                        		
		                        			
		                        			Objective To explore early postoperative gait characteristics and clinical outcomes after total knee arthroplasty(TKA).Methods From February 2023 to July 2023,26 patients with unilateral knee osteoarthritis(KOA)were treated with TKA,including 4 males and 22 females,aged from 57 to 85 years old with an average of(67.58±6.49)years old;body mass in-dex(BMI)ranged from 18.83 to 38.28 kg·m-2 with an average of(26.43±4.15)kg·m-2;14 patients on the left side,12 pa-tients on the right side;according to Kellgren-Lawrence(K-L)classification,6 patients with grade Ⅲ and 20 patients with grade Ⅳ;the courses of disease ranged from 1 to 14 years with an average of(5.54±3.29)years.Images and videos of standing up and walking,walking side shot,squatting and supine kneeling were taken with smart phones before operation and 6 weeks after operation.The human posture estimation framework OpenPose were used to analyze stride frequency,step length,step length,step speed,active knee knee bending angle,stride length,double support phase time,as well as maximum hip flexion angle and maximum knee bending angle on squatting position.Western Ontario and McMaster Universities(WOMAC)arthritis index and Knee Society Score(KSS)were used to evaluate clinical efficacy of knee joint.Results All patients were followed up for 5 to 7 weeks with an average of(6.00±0.57)weeks.The total score of WOMAC decreased from(64.85±11.54)before op-eration to(45.81±7.91)at 6 weeks after operation(P<0.001).The total KSS was increased from(101.19±9.58)before opera-tion to(125.50±10.32)at 6 weeks after operation(P<0.001).The gait speed,stride frequency and stride length of the affected side before operation were(0.32±0.10)m·s-1,(96.35±24.18)steps·min-1,(0.72±0.14)m,respectively;and increased to(0.48±0.11)m·s 1,(104.20±22.53)steps·min-1,(0.79±0.10)m at 6 weeks after operation(P<0.05).The lower limb support time and active knee bending angle decreased from(0.31±0.38)sand(125.21±11.64)° before operation to(0.11±0.04)s and(120.01±13.35)° at 6 weeks after operation(P<0.05).Eleven patients could able to complete squat before operation,13 patients could able to complete at 6 weeks after operation,and 9 patients could able to complete both before operation and 6 weeks after operation.In 9 patients,the maximum bending angle of crouching position was increased from 76.29° to 124.11° before operation to 91.35° to 134.12° at 6 weeks after operation,and the maximum bending angle of hip was increased from 103.70° to 147.25° before operation to 118.61° to 149.48° at 6 weeks after operation.Conclusion Gait analysis technology based on artificial intelligence image recognition is a safe and effective method to quantitatively identify the changes of pa-tients'gait.Knee pain of KOA was relieved and the function was improved,the supporting ability of the affected limb was im-proved after TKA,and the patient's stride frequency,stride length and stride speed were improved,and the overall movement rhythm of both lower limbs are more coordinated.
		                        		
		                        		
		                        		
		                        	
3.Analysis of Thalassemia Gene Mutation Types and Ethnic Distribution Characteristics in Hechi Area,Guangxi
Li-Fang LIANG ; Xiu-Ning HUANG ; Dong-Ming LI ; Bi-Yan CHEN ; Xiang CHEN ; Zhen-Ren PENG ; Sheng HE
Journal of Experimental Hematology 2024;32(4):1191-1196
		                        		
		                        			
		                        			Objective:To investigate the genotype,mutation type,and ethnic distribution characteristics of thalassemia in the population of Hechi area,Guangxi,and to provide a reference basis for prevention and control of thalassemia and eugenic counseling in the region.Methods:Gap-polymerase chain reaction(gap-PCR)and reverse dot blot(RDB)were used for genetic testing on suspected thalassemia persons,and the results were analyzed.Results:Among 29 136 samples,a total of 17 016(58.40%)positive samples for thalassemia genes were detected,with a higher detection rate in males than in females(X2=49.917,P<0.001).The detection rates of thalassemia genes were significant different among Zhuang,Han,Yao,Mulao,and Maonan ethnic groups(x2=546.121,P<0.001).The α-thalassemia genotypes were mainly--SEA/αα(16.67%),-α3.7/αα(8.90%),αCSα/αα(6.00%).Additionally,four rare genotypes were detected,including--THAI/αα(47 cases),HKαα/αα(2 cases),--SEA/-α21.9(2 cases),and--THAI/αcsα(1 case).The β-thalassemia genotypes were mainly βCD17/βN(7.49%),βCD41-42/βN(6.70%),βCD71-72/βN(0.44%).108 cases of moderate and severeβ-thalassemia were detected,of which 81 cases had a history of blood transfusion,the transfusion frequency of 60 cases was more than 10 times/year,and 10 cases received bone marrow transplantation.Conclusion:Thalassemia in Hechi area is predominantly deletion type--SEA/αα,the detection rate of thalassemia in ethnic minorities is higher than that in Han population.In this area,moderate and severe β-thalassemia have certain incidence,these patients mostly need regular blood transfusion and iron removal treatment,and very few patients have received bone marrow transplantation.This study provides a certain reference basis for prevention and control of thalassemia and eugenic counseling in the region.
		                        		
		                        		
		                        		
		                        	
4.Mechanisms of hypertension inducing erectile dysfunction via the cGMP/PKG signaling pathway:An investigation using transcriptomics and network pharmacology
Jun-Long FENG ; Hai-Song LI ; Song SUN ; Bin WANG ; Hua-Nan ZHANG ; Zi-Xiang GAO ; Peng-Ming MAO ; Long-Ji SUN ; Nian-Wen HUANG ; Ji-Sheng WANG
National Journal of Andrology 2024;30(9):771-781
		                        		
		                        			
		                        			Objective:To explore the mechanism of hypertension inducing erectile dysfunction(ED)using transcriptomics and network pharmacology.Methods:We randomly divided 12 male rats with spontaneous hypertension(SHT)into an L-arginine(LA)group(n=6)and an SHT model control(MC)group(n=6),took another 6 Wistar Kyoto male rats as normal controls(NC),and treated the animals in the LA group by intraperitoneal injection of LA at 400 mg/kg and those in the latter two groups with physio-logical saline,once a day,all for 7 days.Then we observed the blood pressure and penile erection of the rats,and determined the ex-pressions of the cGMP/PKG signaling pathway-related proteins and mRNAs in different groups using ELISA,Western blot and RT-qPCR.Results:Transcriptomics combined with network pharmacology showed that the cGMP/PKG signaling pathway played a key role in hypertension-induced ED.In vivo animal experiments revealed a significantly lower frequency of penile erections in the MC than in the NC group(1.33±0.52 vs 2.67±0.51,P<0.05).The protein expressions of eNOS,PKG and sGC were markedly de-creased in the model controls compared with those the normal controls(P<0.05),but remarkably upregulated in the LA group com-pared with those in the MC group(P<0.05).Conclusion:Hypertension decreases the expressions of eNOS,NO,sGC,cGMP and PKG proteins and the level of testosterone by inhibiting the cGMP/PKG signaling pathway,which consequently suppresses the relaxa-tion of the penile vascular smooth muscle and reduces erectile function.
		                        		
		                        		
		                        		
		                        	
5.Clinical characteristics and microbiological tests analysis of 18 patients with Talaromyces marneffei infection
ZOU Sheng-hua ; FANG Mei-lan ; LIN Zhen-li ; CHEN Xin-chao ; HUANG Ming-xiang ; GUAN Hong-lian ; WANG Xin-hang
China Tropical Medicine 2023;23(2):139-
		                        		
		                        			
		                        			Abstract:Objective To investigate the clinical characteristics and early diagnostic methods of patients with Talaromyces marneffei infection, so as to reduce the mortality of patients. Methods    The clinical characteristics and microbiological analysis data including fungal culture, smear examination and mass spectrometry were collected from 18 patients with Talaromyces marneffei infection in the Department of Respiratory Medicine, Department of Tuberculosis, and Department of Critical Respiratory Medicine in Fuzhou Pulmonary Hospital from January 2017 to December 2021, and descriptive analysis was conducted. Results All the 18 patients were confirmed to be infected with Talaromyces marneffei by conventional culture and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS). The main infection sites of 18 patients with Talaromyces marneffei infection were lungs and lymph nodes, and the patients were accompanied by clinical manifestations such as cough, sputum and fever. The imaging features such as patchy shadows, mediastinal lymph node shadows and nodular shadows were common. Microbiological testing showed a statistically significant difference between smear and culture with a higher positive culture rate (χ2=13.74, P<0.05). The positive rate of blood culture in microbiological test was 60.0% (9/15), the positive rate of bronchial lavage fluid culture was 26.7% (4/15), the positive rate of sputum culture was 5.6% (1/18), one case each of pus, bone marrow, pleural fluid and cerebrospinal fluid was positive for culture and the other cases were negative, one case of sputum and one case of pus were positive for smear and the rest were negative. Colony characteristics showed that the colony morphology was mycelial phase at 25 ℃, producing red pigment, and the branching pattern of the penicillus was seen microscopically as monoverticillate or biverticillate; At 35 ℃, the yeast phase appeared at the initial stage, and then the mycelium phase changed after 5-6 days; the yeast phase was observed at 37 ℃, and yeast-like cells were seen under the microscope. All 18 patients with Talaromyces marneffei infection got better after using antifungal drugs. Compared with non-HIV patients with Talaromyces marneffei infection, leukopenia and anemia were common in HIV patients with Talaromyces marneffei infection, and the differences were statistically significant (P<0.05).  Conclusions    The infection of Talaromyces marneffei can be divided into localized type and disseminated type, which usually invade the lungs, skin, lymph nodes and other places. The main manifestations of patients are fever, cough, phlegm and other atypical symptoms. At present, the diagnosis of Talaromyces marneffei infection is mostly based on the fungal culture test, and the application of MALDI-TOF MS method can effectively shorten the diagnosis time of Talaromycosis marneffei. Clinical characteristics combined with microbiological analysis provide an objective basis for early diagnosis of patients with Talaromyces marneffei infection, and timely use of antifungal therapy can improve the prognosis of patients.
		                        		
		                        		
		                        		
		                        	
6.Vasectomy reversal in China during the recent decade: insights from a multicenter retrospective investigation.
Li-Juan REN ; Rui-Zhi XUE ; Zi-Qiang WU ; Er-Lei ZHI ; Wei LI ; Liang HUANG ; Xu-Yu XIANG ; Dao-Yuan LI ; Xue-Ming LIN
Asian Journal of Andrology 2023;25(3):416-420
		                        		
		                        			
		                        			To date, there is little information about the demography of vasectomy reversal (VR) patients or the factors currently influencing VR effectiveness in China, especially after the universal two-child policy was released in 2015. In this research, demographic data and perioperative medical records of VR patients were extracted from seven major hospitals in different provinces or municipalities of China. Meanwhile, a telephone survey of the patients was conducted to collect follow-up information. Eventually, 448 VR cases from the past 13 years were included. The results were analyzed by stratified comparison to investigate factors that can influence postoperative vas deferens patency and pregnancy rate. Appropriately statistical methods were used, and all of the protocols were approved by the Ethics Committees of the institutes in this research. The results showed that the annual operation volume of VR quadrupled after the two-child policy was implemented. Nonmicrosurgery and a long duration of vasectomy were significantly associated with a lower patency rate. A follow-up survey showed that the general postoperative pregnancy rate was 27.2%. For female partners over the age of 35 years, the postoperative pregnancy rate showed a more severe decline, but only 35.5% of them had been given a fertility examination before their husbands' VR surgery. Our work revealed that more patients in China have been demanding VR in recent years. High-quality microsurgery and a short duration of vasectomy are crucial for restoring patency by VR. Clinical andrologists should perform a preoperative fertility evaluation of the patients' female partners.
		                        		
		                        		
		                        		
		                        			Male
		                        			;
		                        		
		                        			Pregnancy
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Adult
		                        			;
		                        		
		                        			Vasovasostomy
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Vas Deferens/surgery*
		                        			;
		                        		
		                        			Vasectomy
		                        			;
		                        		
		                        			China/epidemiology*
		                        			
		                        		
		                        	
7.Research progress of occupational hazards in plywood manufacturing.
Xiang HUANG ; Yue Ming JIANG ; Qing Qing NONG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2023;41(4):317-320
		                        		
		                        			
		                        			Occupational disease hazards in plywood manufacturing mainly include wood dust, formaldehyde, phenol, ammonia, noise, terpene, microorganisms, etc. The exposure is complex with multiple factors accompanied or coexisted. In the production process, these factors are exceeded, and mass occupational disease hazard events occurred among workers. Exposure to wood dust, formaldehyde, terpene, etc., put workers at increased risk of cancer. This article provides a review of this issue in order to provide a scientific basis for the prevention and control of occupational disease hazards in plywood manufacturing.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Wood/chemistry*
		                        			;
		                        		
		                        			Occupational Diseases/chemically induced*
		                        			;
		                        		
		                        			Formaldehyde/adverse effects*
		                        			;
		                        		
		                        			Terpenes
		                        			;
		                        		
		                        			Dust
		                        			;
		                        		
		                        			Occupational Exposure/adverse effects*
		                        			
		                        		
		                        	
8.Discussion on "sugar needle - comfortable acupuncture and moxibustion".
Tao HUANG ; Xiang-Hong JING ; Neng-Gui XU ; Qiang WU ; Yong-Ming LI ; Can-Hui LI ; Ye-Meng CHEN ; Wei-Dong LU ; Yi-Fan YANG ; Tian-Jun WANG ; Shao-Bai WANG
Chinese Acupuncture & Moxibustion 2023;43(3):341-344
		                        		
		                        			
		                        			The cases of feeling comfort during acupuncture and moxibustion treatment in literature were summarized and its biological basis was explored. A simple classification of comfort was made, and the importance of obtaining comfort in acupuncture treatment was pointed out. Considering the pursuit of less pain and harmlessness in modern clinical treatment, sugar needle should be advocated and popularized in current clinical practice of acupuncture and moxibustion.
		                        		
		                        		
		                        		
		                        			Sugars
		                        			;
		                        		
		                        			Moxibustion
		                        			;
		                        		
		                        			Acupuncture Therapy
		                        			;
		                        		
		                        			Emotions
		                        			;
		                        		
		                        			Needles
		                        			
		                        		
		                        	
9.Clinical analysis of inflatable video-assisted mediastinoscopic transhiatal esophagectomy combined with laparoscopy.
Zhi Ning HUANG ; Chang Qing LIU ; Ming Fa GUO ; Mei Qing XU ; Xiao Hui SUN ; Gao Xiang WANG ; Ming Ran XIE
Chinese Journal of Surgery 2023;61(1):48-53
		                        		
		                        			
		                        			Objective: To examine the safety and effectiveness of inflatable video-assisted mediastinoscopic transhiatal esophagectomy (IVMTE). Methods: Totally 269 patients admitted to the Anhui Provincial Hospital of Anhui Medical University who underwent IVMTE (IVMTE group, n=47) or thoracoscopy combined with minimally invasive Mckeown esophageal cancer resection (MIME group, n=222) from September 2017 to December 2021 were analyzed retrospectively. There were 31 males and 16 females in IVMTE group, aged (68.6±7.5) years (range: 54 to 87 years). There were 159 males and 63 females in MIME group, aged (66.8±8.8) years (range: 42 to 93 years). A 1∶1 match was performed on both groups by propensity score matching, with 38 cases in each group. The intraoperative conditions and postoperative complication rates of the two groups were compared by t test, Wilcoxon rank, χ2 test, or Fisher exact probability method. Results: Patients in IVMTE group had less intraoperative bleeding ((96.0±39.2) ml vs. (123.8±49.3) ml, t=-2.627, P=0.011), shorter operation time ((239.1±47.3) minutes vs. (264.2±57.2) minutes, t=-2.086, P=0.040), and less drainage 3 days after surgery (85(89) ml vs. 675(573) ml, Z=-7.575, P<0.01) compared with that of MIME group. There were no statistically significant differences between the two groups in terms of drainage tube-belt time, postoperative hospital stay, and lymph node dissection stations and numbers (all P>0.05). The incidence of Clavien-Dindo grade 1 to 2 pulmonary infection (7.9%(3/38) vs. 31.6%(12/38), χ²=6.728, P=0.009), total complications (21.1%(8/38) vs. 47.4%(18/38), χ²=5.846, P=0.016) and total lung complications (13.2%(5/38) vs. 42.1%(16/38), χ²=7.962, P=0.005) in the IVMTE group were significantly lower. Conclusion: Inflatable video-assisted mediastinoscopic transhiatal esophagectomy combined with laparoscopic esophagectomy is safe and feasible, which can reach the same range of oncology as thoracoscopic surgery.
		                        		
		                        		
		                        		
		                        			Male
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Esophagectomy/methods*
		                        			;
		                        		
		                        			Treatment Outcome
		                        			;
		                        		
		                        			Laparoscopy
		                        			;
		                        		
		                        			Thoracoscopy
		                        			;
		                        		
		                        			Lymph Node Excision/methods*
		                        			;
		                        		
		                        			Esophageal Neoplasms/surgery*
		                        			;
		                        		
		                        			Postoperative Complications
		                        			
		                        		
		                        	
10.Neuroprotective effect and mechanism of cPLA2 inhibitor increases autophagic flux on spinal cord injury.
Wen-Hai YAN ; Ming-Sheng TAN ; Cheng HUANG ; Nan-Shan MA ; Xiang-Sheng TANG
China Journal of Orthopaedics and Traumatology 2023;36(9):873-879
		                        		
		                        			OBJECTIVE:
		                        			To investigate the mechanism of cytosolic phospholipase A2(cPLA2) inhibitor to improve neurological function after spinal cord injury (SCI).
		                        		
		                        			METHODS:
		                        			Thirty-six 3 months old female SD rats, with body mass (280±20) g, were divided into three groups (n=12):sham group, SCI group, and SCI+ arachidonyl trifluoromethyl ketone(AACOCF3) group. Balloon compression SCI model was established in all three groups. In the sham model group, the spinal cord compression model was created after the balloon was placed without pressure treatment, and the remaining two groups were pressurized with the balloon for 48 h. After successful modeling, rats in the SCI+AACOCF3 group were injected intraperitoneally with AACOCF3, a specific inhibitor of cPLA2. The remaining two groups of rats were injected intraperitoneally with saline. The animals were sacrificed in batches on 7 and 14 days after modeling, respectively. And the damaged spinal cord tissues were sampled for pathomorphological observation, to detect the expression of cPLA2 and various autophagic fluxPrelated molecules and test the recovery of motor function.
		                        		
		                        			RESULTS:
		                        			Spinal cord histomorphometry examination showed that the spinal cord tissue in the sham group was structurally intact, with normal numbers and morphology of neurons and glial cells. In the SCI group, spinal cord tissue fractures with large and prominent spinal cord cavities were seen. In the SCI+AACOCF3 group, the spinal cord tissue was more intact than in the SCI group, with more fused spinal cord cavities, more surviving neurons, and less glial cell hyperplasia. Western blot showed that the sham group had the lowest protein expression of LC3-Ⅱ, Beclin 1, p62, and cPLA2 compared with the SCI and SCI+AACOCF3 groups (P<0.05) and the highest protein expression of LC3-Ⅰ (P<0.05). P62 and cPLA2 expression in the SCI group were higher than in the SCI+AACOCF3 group (P<0.05). Behavioral observations showed that the time corresponding to BBB exercise scores was significantly lower in both the SCI and SCI+AACOCF3 groups than in the sham group (P<0.05). Scores at 3, 7, and 14 days after pressurization were higher in the SCI+AACOCF3 group than in the SCI group (P<0.05).
		                        		
		                        			CONCLUSION
		                        			cPLA2 inhibitors can reduce neuronal damage secondary to SCI, promote neurological recovery and improve motor function by improving lysosomal membrane permeability and regulating autophagic flux.
		                        		
		                        		
		                        		
		                        			Female
		                        			;
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Rats
		                        			;
		                        		
		                        			Rats, Sprague-Dawley
		                        			;
		                        		
		                        			Neuroprotective Agents/pharmacology*
		                        			;
		                        		
		                        			Spinal Cord Injuries/drug therapy*
		                        			;
		                        		
		                        			Spinal Cord Compression
		                        			
		                        		
		                        	
            
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