mRNA Expression of Aquaporins in Human Luteinized Granulosa Cells from Different Diameter Follicles / 中山大学学报(医学科学版)

ObjectiveTo investigate the mRNA expression levels of various aquaporins （AQPs） in luteinized granulosa cells from follicles of different diameters. MethodsFrom March 25， 2022 to September 23， 2022 in our reproductive medicine center， 48 women undergoing in-vitro fertilization （IVF） were enrolled and divided into the antagonist group and the agonist group according to the ovarian stimulation protocol. Follicular fluid samples were collected on the day of oocyte pick-up and granulosa cells were extracted from follicles of different diameters： small （<13 mm）， medium （13~18 mm） and large （≥18 mm）. After RNA quantification， 22 cases （66 samples） were included for analysis and mRNA expression levels of AQPs were compared among the three follicle groups. ResultsThe mRNA expression of aquaporin 2 （AQP2） in luteinized granulosa cells increased with the increase of follicle diameter （linear trend P = 0.004） and the difference was statistically significant between two groups of large and small follicles （P = 0.017）. Statistical difference was found in the antagonist group （P = 0.049 6）， but not in the agonist group （P = 0.108）. ConclusionThe mRNA level of AQP2 in luteinized granulosa cells increases with the increase of follicle diameter and its expression is related to the ovarian stimulation protocol， suggesting that AQP2 may play a role in follicle growth and follicular fluid formation， and its mRNA expression level may be regulated by follicle stimulating hormone （FSH） and luteinizing hormone （LH）.

Clinical study on new risk assessment and prediction system for early osteonecrosis of the femoral head / 中国骨伤

OBJECTIVE@#To establish a risk assessment and prediction system for early osteonecrosis of the femoral head (ONFH) in order to predict the collapse risk.@*METHODS@#The risk assessment system for early necrosis and collapse of femoral head was established based on the combination of Steinberg stage, ABC typing and the proportion of the proximal sclerotic rim. Firstly, Steinberg stage system was applied. ABC typing was applied to predict risk in stage I, type C was risk free, type B was low risk, type A and type BC were medium risk, type A-C and type AB were high risk. The classification of proximal sclerotic rim was first applied when the Steinberg stage was Ⅱ-Ⅲ, and type 2 was expected to be low risk. If the classification of proximal sclerotic rimwas type 1, then the ABC typing was applied, type C was risk-free, type B was low risk, type A and type BC were medium risk, type A-C and type AB were high risk. According to this prediction system, the collapse risk of femoral head in 188 cases(301 hips) were predicted by retrospective analysis. All the hips were enrolled at the out-patient department of orthopedic in Guang'anmen Hospital attached to China Academy of Chinese Medical Science. The consistency of the prediction results of three doctors and one doctor at different times were evaluated.@*RESULTS@#Among them, 136 cases were male, 52 were female. 75 cases were single hip, 113 were double hip. The age of the patients wa 19 to 64(42.61±12.07) years. The natural course of disease was 0.33 to 5.00(3.62±1.93) years. 206 hips in 301 hips had collapsed, with a collapse rate of 68.44%. In the risk-free group, none hip had collapsed, with a collapse rate of 0%. In the low-risk group, 9 hip in 91 hips had collapsed, with a collapse rate of 9.89%. In the medium-risk group, 12 hip in 19 hips had collapsed, with a collapse rate of 63.16%. And in the high risk group, 185 hips in 190 hips had collapsed, with a collapse rate of 97.37%. They were significantly differences in their collapse rate (@*CONCLUSION@#The risk assessment and prediction system for early ONFH selects different methods to predict the risk of collapse according to the imaging characteristics of different stages, which is combines with the comprehensive assessment of multiple risk factors. The system is applicable to a wide range, simple operation and convenient for clinical application.

Protective Effects of Scutellarin on Testicular Vessels of Rat with Type II Diabetes Mellitus-Induced Testicular Damages / 中山大学学报(医学科学版)

@#【Objective】To evaluate the protective effect of Scutellarin（SCU）on type II diabetes-induced disorder of testes and show the mechanism of Scutellarin′s action.【Method】We divided 24 Wistar rats equally into 3 groups：control group，diabetes group and diabetes + SCU group. We used streptozotocin and high-fat diet to establish type Ⅱ diabetic rat model on Wistar rat. Haematoxylin and eosin staining were used to evaluate the testicular morphologic changes. Immunohistochemical staining was used to measure the expression level of vascular endothelial growth factor（VEGF）in testes. Oxidative stress in testes and epididymis was tested by fluorescence spectrophotometer and enzyme linked immunosorbent assay（ELISA）. The blood flow rate in testicular vessels was measured by Doppler. 【Results】 Our results showed that hyperglycemia induced morphologic impairments in testes of rats, higher concentrations of Malondialdehyde（MDA） and ROS in testes and epididymis, lower testicular blood flow rate, while administration of Scutellarin could significantly inhibit these damages（DCF：12.93 vs. 7.63 mg/min ；MDA：2.37 vs. 1.48 mg/min ；testicular blood flow rate：110.0 vs. 210.2 mm/s；P < 0.05）.【Conclusion】Our data suggest that SCU has the potential to reverse male reproductive disorder in diabetes by reducing the concentrations of ROS and improving. microcirculatory disturbances.

Effect of Huoxue Ointment on Osteoarthritis Caused by Knee Joint Injury in Rabbits / 中国实验方剂学杂志

Objective:To investigate the efficacy and anti-inflammatory and analgesic effect of Huoxue ointment on osteoarthritis of the knee caused by knee joint injury in rabbits. Method:A total of 60 big ear white rabbits were selected, with half male and half female. Ten rabbits were randomly selected by weight and set as normal control group. For the remaining rabbits, the injury of articular cartilage and anterior cruciate ligament were caused in the femoral ankle joint surface of the right hind knee joint, so as to establish an animal model of knee osteoarthritis in rabbits. For the normal control group, the skin was cut open and immediately sutured. Two weeks after the operation, rabbits were randomly divided into model control group according to body weight，positive voltaren group (1 cm·kg-1·d-1), and high-dose (6 g·kg-1·d-1), medium-dose (3 g·kg-1·d-1) and low-dose(1.5 g·kg-1·d-1) Huoxue ointment groups, with 10 animals in each group. Interleukinin-1β （IL-1β） in rabbit articular cavity fluid and peripheral blood were measured, contents of tumor necrosis factor-α （TNF-α） and free radical nitric oxide（NO）, rabbit articular cartilage was examined by histopathology, in order to study the efficacy of Huoxue ointment on rabbit knee arthritis. The model of increased permeability of capillaries in abdominal cavity of mice induced by acetic acid and the model of foot swelling of rats induced by carrageenan were used to determine the absorbance of mice peritoneal fluid and the rat foot swelling value, and study the anti-inflammatory effect of huoxue ointment. A total of 120 SPF ICR mice and 60 SD rats were selected, with half male and half female. The mice were randomly divided according to body weight into model control groups, positive Voltaren group (3 cm·kg-1·d-1), and high-dose (16 g·kg-1·d-1), medium-dose (8 g·kg-1·d-1), low-dose (4 g·kg-1·d-1) Huoxue ointment groups，with 12 animals in each group. The rats were randomly divided into the model control group, positive voltaren group (2 cm·kg-1·d-1), and high-dose (8 g·kg-1·d-1), medium-dose (4 g·kg-1·d-1), low-dose (2 g·kg-1·d-1) Huoxue ointment groups，with 10 animals in each group. Result:The tested dose of Huoxue ointment，the levels of IL-1β, TNF-α and NO in rabbit serum and joint fluid were decreased to varying degrees, and the effect was significant in the high-dose group (P<0.05). According to the pathological examination, the tested dose of Huoxue ointment, knee cartilage and synovial tissue lesions were significantly lower than those of the model control group (P<0.05)，the value of peritoneal fluid concentration and the foot swelling in medium and high-dose Huoxue ointment groups were significantly lower than those in the model control group (P<0.05). The analgesic effect of tail shaking in mice, and the pain threshold rate of the high-dose Huoxue ointment group were significantly higher than those of the model control group (P<0.05)， the analgesic effect lasted for 4 hours, the foot tenderness of rats in the large and medium-dose Huoxue ointment groups was significantly higher than that in the model control group (P<0.05). Conclusion:Huoxue ointment has obvious joint repair and anti-inflammatory and analgesic effects on osteoarthritis of knee caused by knee joint injury in rabbits.

Application of methods on target identification and validation of label-free natural products / 药学学报

Natural products areone source of new drugs, and their target identification is pivotal forelucidating the mechanism of action. Most methods of target discovery and validation utilize labeling natural products with probes. This is time-consuming and laborious, and often results in activity decrease or change of the natural products. Recent methods withoutchemicalmodificationhave become the main force in the target identification of natural products, including direct or indirect methods. Direct methods are mainly based on the principle of affinity and stability of protein, and indirect methods infer the drug target from the change in physiological responses or biochemical signatures. This review summarizes recent methods for target identification and validation of label-freenatural products, providing new ideas and strategies for future research in natural products.

Clinical Outcome Analysis of Single Embryo Transfer / 中山大学学报(医学科学版)

@#【Objective】To explore the application prospect of single embryo transfer（SET）in assisted reproduction. 【Methods】A total of 5 130 cases of fresh and frozen embryo transfer in the Reproductive Center of the First Affiliated Hospital，Sun Yat-sen University in the recent two years were analyzed retrospectively. To analyze the main outcome of clinical pregnancy in different age groups，we classified the SET cycles into 4 groups of fresh or frozen，elective or obliged.【Results】In non-preimplantation genetic testing（PGT）elective SET cycles，the clinical pregnancy rate of the day 3 embryos（D3）and day 6 embryos（D6）in patients under 35 were as high as 35%. The clinical pregnancy rate of day 5 embryos（D5）was 50% approximately. While in PGT cycles，the clinical pregnancy rate of patients less than 42 years old was 45%，especially implantation rate of D5 embryos was close to 60% in all patients under 40. With the increase of age，the pregnancy rate of SET decreased. In non-PGT group，D3 embryos showed significant difference in either elective/obliged or fresh/frozen cycles（P < 0.001 for all）. In general，elective SET showed a higher pregnancy rate than obliged cycles. There was no significant difference between the elective and obliged frozen cycles of D5 or D6 embryos regarding clinical pregnancy rate（P = 0.074 and P = 0.596，respectively）. Elective D5 embryo transfer group had a higher pregnancy rate than that of the obliged group（P = 0.011）. However，there was no significant difference of clinical pregnancy rate between elective and obliged groups of D6 embryos（P = 0.979）. In non-PGT cycles，clinical pregnancy rate was significantly different among different age groups of either elective/obliged or fresh/ frozen cycles（P < 0.001）. Obliged D3 and D6 embryo transfer groups showed the lowest pregnancy rate，and in PGT cycles of patients under 40，D5 embryo showed a higher pregnancy rate than that of D6 group as a whole（P < 0.001）.【Conclusion】The outcome of SET strategy seemed good in the last 2 years. In order to avoid perinatal complications caused by multiple fetuses，clinicians and patients should actively promote and choose SET.

Case-control study on the treatmentof the fifth metatarsal base fractures by cardboard compression pad versus short leg plaster / 中国骨伤

<p><b>OBJECTIVE</b>To compare the effect,safety,and advantage of flexible fixation with paperboard and pad versus short leg plaster in treating the fifth metatarsal base fracture,and establish the standard of diagnosis and treatment of the fifth metatarsal base fractures in flexible fixation with paperboard and pad.</p><p><b>METHODS</b>From June 2010 to March 2013,59 patients with the fifth metatarsal base fracture were treated with paperboard and pad fixation or short leg plaster. Patients were enrolled and divided into paperboard and pad treatment group (paperboard group) and short leg plaster treatment group (plaster group) randomly according to the random number table. In paperboard group,there were 29 cases including 9 males and 20 females with an average age of (51.79±11.40) years old; the average course of injury was (11.59±6.58) hours. In plaster group, there were 30 cases including 9 males and 21 females with an average age of (52.13+17.34) years old ;the average course of injury was (11.03±7.06) hours. According to whether the fracture line across the articular surface, in paperboard group there were 14 cases of type A,15 of type B; in plaster group,16 of type A, 14 of type B. According to the degree of dislocation,in paperboard group there were 16 cases of degree I ,13 of degree II ; in plaster group,20 were degree I ,10 were degree II. Fracture was restored according to the type in manual. Patients in paperboard group were treated with paperboard and pad, and patients in plaster group were treated with short leg plaster. Fracture was fixed for 4 to 6 weeks according to fracture healing. On the 2nd, 4th,6th, 8th week and 3rd, 6th month after fixation, patients were followed up, and the foot function score was used to evaluate the function of injured foot. X-ray of injured foot was taken on the 2nd, 4th, 6th and 8th week were used to assess fracture healing.</p><p><b>RESULTS</b>All patients got complete follow-up. The X-ray result showed that all fracture reached at clinical healing on the 8th week after fixation without skin ulcer,nonunion and displacement of fracture. From the 4th to 8th week after fixation, paperboard group had a higher X-ray score than plaster group, but the difference between two groups had no statistically significance. Repeated analysis result showed that there was interact at different time point and between groups,the difference had statistically significance (P<0.01). The foot function score showed that at all time point, paperboard group had a higher score than plaster group, and on the 2nd, 4th, and 6th week, it had statistically significant difference(P<0.01) between two groups. On the 6th months after fixation,the excellent and good rate of paperboard group was 93.10%, higher than that of plaster group, which was 86.67%. But it had no statistically difference(P=0.483) between two groups.</p><p><b>CONCLUSION</b>Using paperboard and pad fixation to treat the fifth metatarsal base fracture has the advantage of simplicity operating,reliable fixation, satisfactory effects, easily obtainable material.</p>

Clinical observation of jianpi bushen formula to prevent collapse of osteonecrosis of femoral head / 中国中药杂志

To observe the clinical effect of Jianpi Bushen formula to prevent the collapse of osteonecrosis of femoral head (ONFH) of type phlegm and blood stasis obstructing the collaterals. 50 cases (including 73 hips) of non-collapse ONFH (ARCO I, II, III a) were selected from the out-patient department of orthopedic in Guang'anmen Hospital attached to China Academy of Chinese Medical Science. All the cases fit for diagnostic criteria were given Jianpi Bushen formula and followed up. Staging criteria was ARCO classification. Harris score evaluated the hip function. The mean follow-up time was 4.2 years (3-5.4 years). After a mean of 4.2 years (3-5.4 years) followed-up, 12 hips collapse occurred while 61 hips not (the non-collapse rate was 83.56%). Collapse did not occurred among the all 7 hips at stage ARCO I (the non-collapse rate was 100%). Among the 49 hips at stage ARCO II, 9 hips collapse occurred while 40 hips not (the non-collapse rate was 81.63%). Among the 17 hips at stage ARCO III a, 3 hips collapse occurred while 14 hips not (the non-collapse rate was 82.35%). Kaplan-Meier analysis showed the average survival time of non-collapse was 5 years (4. 8-5.2 years). The 3 year survival rate of non-collapse was 92.5%. The 4 year survival rate was 74% and the condition tended to be stability trend. The Harris score was 71.93 +/- 11.25 before treatment and 81.63 +/- 12.16 after treatment, significantly different. These results suggest that: Jianpi Bushen formula is an effective method for treating ONFH of type phlegm and blood stasis obstructing the collaterals. It can delay or prevent the collapse of femoral head and significantly improve the hip function.

Research on the distribution difference of MRI signals in osteonecrosis of the femoral head patients of different TCM syndrome types / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the distribution difference of magnetic resonance imaging (MRI) signals in osteonecrosis of the femoral head (ONFH) patients of different TCM syndrome types.</p><p><b>METHODS</b>Recruited 29 ONFH patients were assigned to the tendon and vessel stagnation group (14 cases) and the Gan-Shen deficiency group (15 cases) according to TCM syndrome typing. The distribution difference of their MRI signals of MRIT1WI, T2WI + fs, T1 and T2 combined signal were compared.</p><p><b>RESULTS</b>There was statistical difference in T1W1 signal distribution between the two TCM syndrome types (P = 0.04). There was no statistical difference in T2WI + fs or T1 and T2 combined signal between the two TCM syndrome types (P = 0.42, P = 0.15). MRI signals in the tendon and vessel stagnation group were mainly manifested as fat-like signals,while they were mainly manifested as mixed signals in the Gan-Shen deficiency group.</p><p><b>CONCLUSION</b>Distribution difference of MRI signals exists between ONFH patients of different TCM syndrome types.</p>

Study on the relationship between clinical syndromes and X-ray manifestation of knee osteoarthritis / 中国骨伤

<p><b>OBJECTIVE</b>To study the relationship between clinical syndromes and X-ray manifestation of knee osteoarthritis (KOA), so as to provide evidence for clinical diagnosis and treatment.</p><p><b>METHODS</b>Seventy-eight patients (108 knees) with KOA from out-patient clinic of orthopedic were collected, aged from 41 to 77, including 65 females (89 knees) and 13 males (19 knees). Questionnair investigation of Lequesne index was performed among all these patients and the Lequesne index was calculated. And all these patients were divided into two groups, mild group (Lequesne index < or = 8) and severe group (Lequesne index > 8). Weight-bearing anteriorposterior and lateral plain film of knee joint and patellofemoral Skyline plain flim was taken. Joint space narrowing, osteophyte formation, subchondral osteosclerosis and subchondral cystic degeneration were evaluated. The lower limb alignment and lateral patella angle were detected. The correlation of the Lequesne index and X-ray manifestation was analyzed by logistic regression analysis.</p><p><b>RESULTS</b>There were significant differences between mild and severe groups in the following indexes: lateral tibiofemoral joint space narrowing, the medial retropatellar space narrowing, medial and lateral tibia osteophytes, medial and tibial femur femur osteophytes, medial and lateral trochlea osteophytes, lateral patella osteophytes, tibia introcondylar osteophytes (P<0.05). The results of the logistic regression analysis showed that the lateral femur osteophytes were the most important radiologic manifestation to evaluate the severity degree of KOA,with the minimum associated probability (0.009) and the maximum wald value (6.779).</p><p><b>CONCLUSION</b>When evaluating the severity degree of KOA,the joint space narrowing and osteophytes are the most sigenificant radiologic manifestation which includs the lateral tibiofemoral joint space narrowing, the medial retropatellar space narrowing, medial and lateral tibia osteophytes, medial and lateral femur osteophytes, medial and lateral trochlea osteophytes, lateral patella osteophyte, tibial intercondylar osteophyte.</p>

Modified vitrification method for cryopreservation of human ovarian tissues / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Vitrification is a prospective technology in ovarian tissue cryopreservation, but it is still in an initial stage. This study was conducted to investigate a modified vitrification protocol for human ovarian tissue, which can be used as an alternative to preserve fertility for young women with cancer who have to undergo cytotoxic therapy and sterilization.</p><p><b>METHODS</b>Ovarian tissue samples were collected from 15 patients and randomly allocated to groups of fresh, vitrification, and conventional slow freezing. A modified carrierless vitrification method was applied. The proportion of morphologically intact follicles in fresh ovarian tissues was compared with that in warmed/thawed tissues. The initial growth of the follicles and the concentrations of estradiol and progesterone were detected to determine the viability and endocrine function of the cryopreserved tissues.</p><p><b>RESULTS</b>The proportion of morphologically intact primordial follicles in the fresh group (97.6%) was significantly higher than that in the other two groups (vitrification group 80.3% and slow-freezing group 72.6%, P < 0.001). In both the vitrification and slow-freezing groups, estradiol and progesterone were secreted continuously during 2-week culture in vitro, the proportion of primary follicles were both significantly increased compared to the fresh group. No statistically significant differences existed between the two groups after cryopreservation in the proportion of both primordial and primary follicles, and the concentrations of estradiol and progesterone (P > 0.05).</p><p><b>CONCLUSION</b>The modified vitrification method for cryopreservation of human ovarian tissues is effective, simple, and inexpensive.</p>

Mononostril-septum-transsphenoidal approach for pituitary adenoma / 中南大学学报(医学版)

OBJECTIVE@#To summarize the mononostril-septum-transsphenoidal approach for pituitary adenoma.@*METHODS@#The clinical features, operative techniques, and outcome of 36 patients with pituitary adenoma were analyzed retrospectively.@*RESULTS@#Tumors were totally removed in 28 cases, and subtotally resected in 8 patients. No patient died after the operation. Endocrine symptom of 31 patients returned to the normal level, the symptom of the other 5 cases were improved. Thirty patients with visual field defects recovered after the operation. Cerebrospinal fluid leakage occurred in one patient, and was cured with conservative treatment in 2 weeks.@*CONCLUSION@#Mononostril-septum-transsphenoidal approach can make use of the natural space of the nasal cavity, which has many advantages, such as direct approach, short operative time, minimal invasion, and few complications. It is a effective transsphenoidal surgical approach.