1.Stability study of umbilical cord mesenchymal stem cells formulation in large-scale production
Wang-long CHU ; Tong-jing LI ; Yan SHANGGUAN ; Fang-tao HE ; Jian-fu WU ; Xiu-ping ZENG ; Tao GUO ; Qing-fang WANG ; Fen ZHANG ; Zhen-zhong ZHONG ; Xiao LIANG ; Jun-yuan HU ; Mu-yun LIU
Acta Pharmaceutica Sinica 2024;59(3):743-750
Umbilical cord mesenchymal stem cells (UC-MSCs) have been widely used in regenerative medicine, but there is limited research on the stability of UC-MSCs formulation during production. This study aims to assess the stability of the cell stock solution and intermediate product throughout the production process, as well as the final product following reconstitution, in order to offer guidance for the manufacturing process and serve as a reference for formulation reconstitution methods. Three batches of cell formulation were produced and stored under low temperature (2-8 ℃) and room temperature (20-26 ℃) during cell stock solution and intermediate product stages. The storage time intervals for cell stock solution were 0, 2, 4, and 6 h, while for intermediate products, the intervals were 0, 1, 2, and 3 h. The evaluation items included visual inspection, viable cell concentration, cell viability, cell surface markers, lymphocyte proliferation inhibition rate, and sterility. Additionally, dilution and culture stability studies were performed after reconstitution of the cell product. The reconstitution diluents included 0.9% sodium chloride injection, 0.9% sodium chloride injection + 1% human serum albumin, and 0.9% sodium chloride injection + 2% human serum albumin, with dilution ratios of 10-fold and 40-fold. The storage time intervals after dilution were 0, 1, 2, 3, and 4 h. The reconstitution culture media included DMEM medium, DMEM + 2% platelet lysate, 0.9% sodium chloride injection, and 0.9% sodium chloride injection + 1% human serum albumin, and the culture duration was 24 h. The evaluation items were viable cell concentration and cell viability. The results showed that the cell stock solution remained stable for up to 6 h under both low temperature (2-8 ℃) and room temperature (20-26 ℃) conditions, while the intermediate product remained stable for up to 3 h under the same conditions. After formulation reconstitution, using sodium chloride injection diluted with 1% or 2% human serum albumin maintained a viability of over 80% within 4 h. It was observed that different dilution factors had an impact on cell viability. After formulation reconstitution, cultivation in medium with 2% platelet lysate resulted in a cell viability of over 80% after 24 h. In conclusion, the stability of cell stock solution within 6 h and intermediate product within 3 h meets the requirements. The addition of 1% or 2% human serum albumin in the reconstitution diluent can better protect the post-reconstitution cell viability.
2.Epidemiological Survey of Hemoglobinopathies Based on Next-Generation Sequencing Platform in Hunan Province, China.
Hui XI ; Qin LIU ; Dong Hua XIE ; Xu ZHOU ; Wang Lan TANG ; De Guo TANG ; Chun Yan ZENG ; Qiong WANG ; Xing Hui NIE ; Jin Ping PENG ; Xiao Ya GAO ; Hong Liang WU ; Hao Qing ZHANG ; Li QIU ; Zong Hui FENG ; Shu Yuan WANG ; Shu Xiang ZHOU ; Jun HE ; Shi Hao ZHOU ; Fa Qun ZHOU ; Jun Qing ZHENG ; Shun Yao WANG ; Shi Ping CHEN ; Zhi Fen ZHENG ; Xiao Yuan MA ; Jun Qun FANG ; Chang Biao LIANG ; Hua WANG
Biomedical and Environmental Sciences 2023;36(2):127-134
OBJECTIVE:
This study was aimed at investigating the carrier rate of, and molecular variation in, α- and β-globin gene mutations in Hunan Province.
METHODS:
We recruited 25,946 individuals attending premarital screening from 42 districts and counties in all 14 cities of Hunan Province. Hematological screening was performed, and molecular parameters were assessed.
RESULTS:
The overall carrier rate of thalassemia was 7.1%, including 4.83% for α-thalassemia, 2.15% for β-thalassemia, and 0.12% for both α- and β-thalassemia. The highest carrier rate of thalassemia was in Yongzhou (14.57%). The most abundant genotype of α-thalassemia and β-thalassemia was -α 3.7/αα (50.23%) and β IVS-II-654/β N (28.23%), respectively. Four α-globin mutations [CD108 (ACC>AAC), CAP +29 (G>C), Hb Agrinio and Hb Cervantes] and six β-globin mutations [CAP +8 (C>T), IVS-II-848 (C>T), -56 (G>C), beta nt-77 (G>C), codon 20/21 (-TGGA) and Hb Knossos] had not previously been identified in China. Furthermore, this study provides the first report of the carrier rates of abnormal hemoglobin variants and α-globin triplication in Hunan Province, which were 0.49% and 1.99%, respectively.
CONCLUSION
Our study demonstrates the high complexity and diversity of thalassemia gene mutations in the Hunan population. The results should facilitate genetic counselling and the prevention of severe thalassemia in this region.
Humans
;
beta-Thalassemia/genetics*
;
alpha-Thalassemia/genetics*
;
Hemoglobinopathies/genetics*
;
China/epidemiology*
;
High-Throughput Nucleotide Sequencing
3.Phenotypes and ATP7B gene variants in 316 children with Wilson disease.
Zhi Kun LU ; Jing CHENG ; Si Min LI ; Yun Ting LIN ; Wen ZHANG ; Xiu Zhen LI ; Hui Ying SHENG ; Xiao Jian MAO ; Hui Fen MEI ; Rui Dan ZHENG ; Cui Li LIANG ; Min Yan JIANG ; Yong Lan HUANG ; Li LIU ; Chun Hua ZENG
Chinese Journal of Pediatrics 2022;60(4):317-322
Objectives: To summarize the clinical phenotypes and the variation spectrum of ATP7B gene in Chinese children with Wilson's disease (WD) and to investigate their significance for early diagnosis. Methods: Retrospective analysis was performed on the clinical data of 316 children diagnosed as WD in Guangzhou Women and Children's Medical Center during the period from January 2010 to June 2021. The general situations, clinical manifestations, lab test results, imaging examinations, and ATP7B gene variant characteristics were collected. The patients were divided into asymptomatic WD group and symptomatic WD group based on the presence or absence of clinical symptoms at the time that WD diagnosis was made. The χ2 test, t test or Mann-Whitney U test were used to compare the differences between groups. Results: Among the 316 children with WD, 199 were males and 117 were females, with the age of 5.4 (4.0, 7.6) years at diagnosis; 261 cases (82.6%) were asymptomatic with the age of 4.9 (3.9, 6.4) years; whereas 55 cases (17.4%) were symptomatic with the age of 9.6 (7.3, 12.0) years. The main symptoms invloved liver, kidney, nervous system, or skin damage. Of all the patients, 95.9% (303/316) had abnormal liver function at diagnosis; 98.1% (310/316) had the serum ceruloplasmin lever lower than 200 mg/L; 97.7% (302/309) had 24-hour urine copper content exceeding 40 μg; only 7.4% (23/310) had positive corneal K-F rings, 8.2% (23/281) had abnormal MRI signals in the lenticular nucleus, and all of them had symptoms of damage in liver, kidney or nervous system. Compared with the group of symptomatic WD, asymptomatic group had higher levels of serum alanine aminotransferase and lower levels ceruloplasmin and 24-hour urine copper [(208±137) vs. (72±78) U/L, (55±47) vs. (69±48) mg/L, 103 (72, 153) vs. 492 (230, 1 432) μg; t=9.98, -1.98, Z=-4.89, all P<0.001]. Among the 314 patients completing genetic sequencing, a total of 107 mutations in ATP7B gene were detected, of which 10 are novel variants, and 3 cases (1.0%) had large heterozygous deletion (exons 10 to exon 11) in ATP7B gene. The percentage of missense mutation in asymptomatic WD children was significantly higher than that in symptomatic WD (81.5% (422/518) vs. 69.1% (76/110), χ²=8.47, P<0.05). WD patients carrying homozygous variant of c.2 333G>T had significantly low levels of ceruloplasmin than those not carrying this variant ((23±5) vs. (61±48) mg/L, t=-2.34, P<0.001). Conclusions: The elevation of serum ALT is an important clue for early diagnosis of WD in children, while serum ceruloplasmin and 24-hour urine copper content are specific markers for early diagnosis of WD. In order to confirm the diagnosis of WD, it is necessary to combine the Sanger sequencing with multiplex ligation-dependent probe amplification or other testing technologies.
Ceruloplasmin/metabolism*
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Child
;
Child, Preschool
;
Copper/metabolism*
;
Copper-Transporting ATPases/genetics*
;
Female
;
Hepatolenticular Degeneration/genetics*
;
Humans
;
Male
;
Mutation
;
Phenotype
;
Retrospective Studies
4.Prognostic analysis of continuous lumbar cistern external drainage after aneurysmal subarachnoid hemorrhage
Shukai LIN ; Gang LI ; Fen ZHOU ; Hui WANG ; Jianfeng ZENG ; Shihe XIAO
International Journal of Surgery 2022;49(6):421-427,F5
Objective:To analyze the influencing factors, prevention and treatment strategy of short-term poor prognosis of continuous lumbar cistern external drainage after aneurysms subarachnoid hemorrhage (aSAH).Methods:Used retrospective research method, the clinical data of 300 patients with aSAH combined with continuous lumbar cistern external drainage treated in Sanya Central Hospital (Hainan Third People′s Hospital) from March 2019 to March 2021 were selected as the training set. In addition, the clinical data of 144 patients with aSAH with continuous lumbar cistern external drainage treated in Sanya Central Hospital (Hainan Third People′s Hospital) from March 2017 to May 2019 were selected as the verification set. According to the results of postoperative follow-up, the patients in the training set were divided into two groups: good prognosis group ( n=208) and poor prognosis group ( n=92). The demographic characteristics, past history, Hunt-Hess grade, modified Fisher grade, location of responsible aneurysm, postoperative complications, bone flap decompression and lumbar cistern drainage were compared between the two groups. The independent risk factors for prognosis of aSAH patients undergoing continuous lumbar cistern external drainage were screened by Cox proportional hazard regression model, and these factors were included and XGboost model was established. The prediction model was validated internally and externally in the training set and verification set: AUROC(C-index) was used to verify the model differentiation; GiViTI calibration band and Hosmer-Lemeshow test were used to verify the model calibration; DCA curve was used to verify the clinical validity of the model. Results:Hunt-Hess grade, modified Fisher grade, drainage duration, average daily drainage volume, shunt-dependent hydrocephalus, aneurysm rebleeding, cerebral vasospasm and delayed cerebral ischemia were independent risk factors for poor prognosis in patients with aSAH who underwent continuous lumbar cistern external drainage( P<0.05). The XGboost model was successfully established by incorporating the above independent risk factors, and the internal and external verification of the XGboost model was carried out in the training set and verification set, respectively, the area under the curve of receiver operating characteristic was 0.882(95% CI: 0.820-0.955) and 0.878(95% CI: 0.774-0.928) respectively, and the model differentiation was good; the 80%-90% confidence interval of the GiViTI calibration curve did not cross the 45° angle bisector ( P>0.05). In the Hosmer-Lemeshow goodness-of-fit test, the P value were 0.581 and 0.716, respectively. The threshold probability value in the DCA curve was 30.4%. The clinical net benefit rate of the training set and verification set were 31% and 34%, respectively, indicating that the prediction model was clinically effective. Conclusions:The independent risk factors for poor prognosis of aSAH patients undergoing continuous lumbar cistern drainage are Hunt-Hess grade, modified Fisher grade, cerebral vasospasm, delayed cerebral ischemia and shunt-dependent hydrocephalus. The XGboost model constructed in this study can effectively predict the prognosis of patients with aSAH undergoing continuous lumbar cistern drainage, and provide reference for the formulation of follow-up treatment plans.
5.Delivery room resuscitation and short-term outcomes of extremely preterm and extremely low birth weight infants: a multicenter survey in North China.
Shuai-Jun LI ; Qi FENG ; Xiu-Ying TIAN ; Ying ZHOU ; Yong JI ; Yue-Mei LI ; Shu-Fen ZHAI ; Wei GUO ; Fang ZHANG ; Rong-Xiu ZHENG ; Hai-Ying HE ; Xia LIU ; Jun-Yi WANG ; Hua MEI ; Hong-Yun WANG ; Hua XIE ; Chao-Mei ZENG ; Li MA ; Ping-Ping ZHANG ; Jin-Yu LI ; Xiao-Ying WANG ; Li-Hua LI ; Hong CUI ; Shu-Lan YANG ; Lu CHEN ; Xiao-Hong GU ; Yan-Ju HU ; Sheng-Shun QUE ; Li-Xia SUN ; Ming YANG ; Wen-Li ZHAO ; Qiu-Yan MA ; Hai-Juan WANG ; Jiu-Ye GUO
Chinese Medical Journal 2021;134(13):1561-1568
BACKGROUND:
Delivery room resuscitation assists preterm infants, especially extremely preterm infants (EPI) and extremely low birth weight infants (ELBWI), in breathing support, while it potentially exerts a negative impact on the lungs and outcomes of preterm infants. This study aimed to assess delivery room resuscitation and discharge outcomes of EPI and ELBWI in China.
METHODS:
The clinical data of EPI (gestational age [GA] <28 weeks) and ELBWI (birth weight [BW] <1000 g), admitted within 72 h of birth in 33 neonatal intensive care units from five provinces and cities in North China between 2017 and 2018, were analyzed. The primary outcomes were delivery room resuscitation and risk factors for delivery room intubation (DRI). The secondary outcomes were survival rates, incidence of bronchopulmonary dysplasia (BPD), and risk factors for BPD.
RESULTS:
A cohort of 952 preterm infants were enrolled. The incidence of DRI, chest compressions, and administration of epinephrine was 55.9% (532/952), 12.5% (119/952), and 7.0% (67/952), respectively. Multivariate analysis revealed that the risk factors for DRI were GA <28 weeks (odds ratio [OR], 3.147; 95% confidence interval [CI], 2.082-4.755), BW <1000 g (OR, 2.240; 95% CI, 1.606-3.125), and antepartum infection (OR, 1.429; 95% CI, 1.044-1.956). The survival rate was 65.9% (627/952) and was dependent on GA. The rate of BPD was 29.3% (181/627). Multivariate analysis showed that the risk factors for BPD were male (OR, 1.603; 95% CI, 1.061-2.424), DRI (OR, 2.094; 95% CI, 1.328-3.303), respiratory distress syndrome exposed to ≥2 doses of pulmonary surfactants (PS; OR, 2.700; 95% CI, 1.679-4.343), and mechanical ventilation ≥7 days (OR, 4.358; 95% CI, 2.777-6.837). However, a larger BW (OR, 0.998; 95% CI, 0.996-0.999), antenatal steroid (OR, 0.577; 95% CI, 0.379-0.880), and PS use in the delivery room (OR, 0.273; 95% CI, 0.160-0.467) were preventive factors for BPD (all P < 0.05).
CONCLUSION
Improving delivery room resuscitation and management of respiratory complications are imperative during early management of the health of EPI and ELBWI.
Birth Weight
;
Bronchopulmonary Dysplasia
;
China/epidemiology*
;
Delivery Rooms
;
Female
;
Gestational Age
;
Humans
;
Infant
;
Infant, Extremely Low Birth Weight
;
Infant, Extremely Premature
;
Infant, Newborn
;
Male
;
Pregnancy
7.Acupuncture at acupoints along the meridians for primary insomnia: a multi-center randomized controlled trial.
Xue-Fen WU ; Xue-Na ZHENG ; Yan WANG ; Xiao-Li CHEN ; Li LIU ; Xin-Ran WEI ; Zeng-Hui YUE
Chinese Acupuncture & Moxibustion 2020;40(5):465-471
OBJECTIVE:
To compare the clinical effect differences among different acupoint selection methods for primary insomnia.
METHODS:
A total of 333 patients with primary insomnia were recruited from 3 study centers and randomly divided into a group A (111 cases, 7 cases dropped off), a group B (111 cases, 5 cases dropped off) and a group C (111 cases, 2 cases dropped off). The patients in the group A were treated with acupuncture at Shenmen (HT 7) and Baihui (GV 20), the patients in the group B were treated with acupuncture at Sanyinjiao (SP 6) and Baihui (GV 20), and the patients in the group C were treated with acupuncture at non-acupoint and Baihui (GV 20). All the treatment was given once a day, 30 min each time; 5 treatments were taken as a course and 5 courses of treatment were given. The Pittsburgh sleep quality index (PSQI) and Athens insomnia scale (AIS) scores were evaluated before and after treatment as well as 4 weeks after treatment. The encephalofluctuograph technology (ET) was observed before and after treatment.
RESULTS:
Compared before treatment, the PSQI scores after treatment and at follow-up were significantly decreased in three groups (<0.01), and the decrease in the group A and the group B was greater than that in the group C (<0.01). Compared before treatment, the AIS scores after treatment and at follow-up was significantly decreased in three groups (<0.01), and the decrease in the group A was greater than that in the group C (<0.05). The interclass and between-groups ET before and after treatment had no significant difference (>0.05).
CONCLUSION
The acupuncture at acupoints along the meridians could improve the sleep quality in patients with primary insomnia, and the therapeutic effect of acupoint along the meridians is better than that of non-acupoint.
Acupuncture Points
;
Acupuncture Therapy
;
Humans
;
Meridians
;
Sleep Initiation and Maintenance Disorders
;
therapy
;
Treatment Outcome
8.Therapeutic observation of acupuncture plus navel acupuncture for urinary retention after radical hysterectomy for cervical cancer
Juan XIANG ; Xu-Hong LI ; Yan-Hua ZHOU ; Shuo YANG ; Xiao-Ling ZENG ; Fen XIE
Journal of Acupuncture and Tuina Science 2019;17(4):245-252
Objective: To observe the clinical efficacy of acupuncture plus navel acupuncture for patients with urinary retention after radical hysterectomy for cervical cancer. Methods: A total of 64 patients with urinary retention after radical hysterectomy for cervical cancer was divided into a navel acupuncture group (22 cases), an acupuncture group (18 cases) and an acupuncture plus navel acupuncture group (24 cases). All three groups received bladder function training and neuromuscular electrical stimulation. In addition, navel points were combined in the navel acupuncture group. Electroacupuncture was conducted to Qihai (CV 6), Zhongji (CV 3), Dahe (KI 12), Shuidao (ST 28), Ciliao (BL 32) and Huiyang (BL 35) in the acupuncture group. The acupuncture plus navel acupuncture group received both treatments. The catheter was removed after 3 d of treatment. Spontaneous urination, residual urine volume, urinary catheter dependence and recurrence after 3 d, 6 d and 9 d of treatment in each group were observed, respectively. Results: In the acupuncture plus navel acupuncture group, the markedly effective rates after 3 d, 6 d and 9 d of treatment were significantly higher than those in the navel acupuncture group and the acupuncture group; the urinary catheter dependence was lower than that of the other two groups, and the differences were statistically significant (P<0.05, P<0.01); the spontaneous urination time was shorter than that of the navel acupuncture group and the acupuncture group, and the differences were statistically significant (P<0.05, P<0.01); the residual urine volume was significantly less than that of the navel acupuncture group and the acupuncture group, and the differences were statistically significant (both P<0.01). After the catheter was removed, recurrence was observed from the next day after spontaneous urination was resumed. There were 2 cases of recurrence in the navel acupuncture group, 2 cases in the acupuncture group and 1 case in the acupuncture plus navel acupuncture group. The recurrence rate of the acupuncture plus navel acupuncture group was significantly lower than that of the navel acupuncture group and the acupuncture group (both P<0.01). Conclusion: Acupuncture plus navel acupuncture has satisfactory efficacy for urinary retention after radical hysterectomy for cervical cancer. It can significantly shorten the urinary retention time, reduce the patient's dependence on urinary catheter, and reduce the residual urine volume.
9. Validation of fatty liver index and hepatic steatosis index for screening of non-alcoholic fatty liver disease in adults with obstructive sleep apnea hypopnea syndrome
Li-Da CHEN ; Jie-Feng HUANG ; Qing-Shi CHEN ; Guo-Fu LIN ; Hui-Xue ZENG ; Xiao-Fen LIN ; Xue-Jun LIN ; Li LIN ; Qi-Chang LIN
Chinese Medical Journal 2019;132(22):2670-2676
Background:
Obstructive sleep apnea hypopnea syndrome (OSAHS) is a contributing factor for non-alcoholic fatty liver disease (NAFLD). Non-invasive algorithms including fatty liver index (FLI) and hepatic steatosis index (HSI) have been used as a screening test for NAFLD in epidemiologic studies. The aim of this study is to compare the diagnostic accuracy of FLI and HSI for NAFLD detection in adults with OSAHS.
Methods:
We enrolled consecutive adult subjects who were newly diagnosed with OSAHS from March 2016 to January 2018. NAFLD was diagnosed by ultrasonography. The accuracy and cut-off point of the FLI and HSI to detect NAFLD were assessed by analyzing the area under the receiver operating characteristic (AUROC) curve and the maximum Youden index analysis, respectively.
Results:
The 326 subjects were diagnosed as NAFLD according to ultrasound findings, while 105 subjects who had normal abdominal ultrasonography were grouped as controls. Both FLI and HSI values were significantly higher in patients with NAFLD compared with controls. The AUROC of FLI and HSI for predicting NAFLD was 0.802 (95% confidence interval [CI] 0.762-0.839) and 0.753 (95% CI 0.710-0.793), respectively. The AUROC of FLI was significantly higher than that of HSI (
10.Clinical observation of rapid massage at Shuidao (ST 28) to prevent postpartum urinary retention.
Feng-Qin GUO ; Lu-Qi WEI ; Shu-Zeng ZHANG ; Xiao-Fen ZHANG
Chinese Acupuncture & Moxibustion 2019;39(3):263-266
OBJECTIVE:
To explore the clinical effect of rapid massage at Shuidao (ST 28) to prevent puerpera from postpartum urinary retention.
METHODS:
A total of 200 puerpera giving birth through vagina were enrolled and divided into an observation group and a control group according to the random number table method, 100 cases in each group. In the observation group, rapid massage at Shuidao (ST 28) was applied. In the control group,there was no intervention and urinated naturally. The traditional Chinese medicine syndrome scale was used to evaluate poor sense of urination, and record puerpera with or without postpartum urinary retention, the poor sense of first urination, the first time of urination, the amount of first urination and postpartum hemorrhage.
RESULTS:
The poor sense of first urination in the observation group was significantly lower than that in the control (<0.001), the first time of urination in the observation group was significantly earlier than that in the control group (<0.001), the amount of first urination in the observation group was significantly more than that in the control group (<0.001), the observation group was superior to the control group to prevent postpartum urinary retention (<0.05).
CONCLUSION
Rapid massage at Shuidao (ST 28) can reduce the poor sense of first urination, significantly advance the time of spontaneous urination, significantly increase the amount of first urination and effectively prevent postpartum urinary retention.
Acupuncture Points
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Female
;
Humans
;
Massage
;
Postpartum Period
;
Pregnancy
;
Urinary Retention
;
therapy
;
Urination

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