Consensus recommendations from Chinese experts on the standard operation procedure of dental operative microscope in endodontics and operative dentistry / 中华口腔医学杂志

Dental operative microscope has been wildly used in endodontics and operative dentistry for many years. Many progresses have been made by using operative microscope in improving the outcomes of root canal therapy, endodontic surgery and operative dentistry. However, in clinical practice, improper use of the dental operative microscope is not uncommon, the reason related to which is the operator's lack of understanding and mastering not only the properties but also the standard operative procedure of the dental operative microscope. To this end, in October 2019, the vice chairman of the Society of Cariology and Endodontology, Chinese Stomatological Association, professor Liang Jingping, organized a group of professional experts in this field, convened a meeting about the standard operative procedure of dental operative microscope. Experts at the meeting had a very heated discussion and the consensuses were reached.

Safety and efficacy of microvascular decompression in elderly patients with cranial neuropathy / 中华老年医学杂志

Objective To investigate the safety and clinical efficacies of microvascular decompression(MVD) on cranial neuropathy such as trigeminal neuralgia (TN),hemifacial spasm (HFS)and glossopharyngeal neuralgia(GN)in elderly patients.Methods Clinical data of 72 patients aged 70 years and over diagnosed as cranial neuropathy and undergoing microvascular decompression in our department from January 2015 to August 2017 were retrospectively analyzed.The curative effect and complications were analyzed after treatment.Results All patients successfully underwent MVD under general anesthesia.The facial pain completely disappeared or was significantly improved in 85.7 ％ (30/35)of TN patients one year after operation.The intermittent tonic contractions of the facial muscles were completely disappeared in 94.3％ (33/35)of HFS patients.Clinical symptoms were completely disappeared in 2 patients with GN,4 patients with TN and HFS and 1 patients with TN and GN.Conclusions The surgery goes well and the patients have good postoperative recovery,with no severe complications such as intracranial hematoma,infarction,cerebrospinal fluid leakage,acute hydrocephalus and death.After an adequate preoperative evaluation,MVD is safe and effective with a few complications.It can significantly improve the quality of life in elderly patients with trigeminal neuralgia,hemifacial spasm,and glossopharyngeal neuralgia.

Protocol for chromosome-specific probe construction using PRINS, micromanipulation and DOP-PCR techniques

ABSTRACT Chromosome-specific probes have been widely used in molecular cytogenetics, being obtained with different methods. In this study, a reproducible protocol for construction of chromosome-specific probes is proposed which associates in situ amplification (PRINS), micromanipulation and degenerate oligonucleotide-primed PCR (DOP-PCR). Human lymphocyte cultures were used to obtain metaphases from male and female individuals. The chromosomes were amplified via PRINS, and subcentromeric fragments of the X chromosome were microdissected using microneedles coupled to a phase contrast microscope. The fragments were amplified by DOP-PCR and labeled with tetramethyl-rhodamine-5-dUTP. The probes were used in fluorescent in situ hybridization (FISH) procedure to highlight these specific regions in the metaphases. The results show one fluorescent red spot in male and two in female X chromosomes and interphase nuclei.

Mcrosurgical reconstructions of hepatic arteries in pediatric liver transplantation performed by a single surgeon:115 cases report / 中华器官移植杂志

Objective Hepatic artery (HA) reconstruction is one challenging procedure in pediatric liver transplantation (PLT).Here we review the first 115 microsurgical reconstructions of HA in PLT performed by a single surgeon,aiming to demonstrate the learning curve and the problems encountered.Methods From July 2016 to January 2017,a series of 115 microsurgical reconstructions of HA in PLT for end-stage liver disease were finished by one single surgeon with 4-year liver surgery experience and 2-week microsurgical training.HA reconstruction was performed with an operating microscope (Carl-Zeiss S88).Reconstruction was completed with interrupted sutures with 8-0 or 9-0 Prolene using the double clip for fixation.The blood flow was examined by Doppler scan daily after PLTs in first week and then once in 2nd week and first month for patency.A total of 143 artery anastomoses were performed in 115 PLTs.The age ranged from 3 months to 9 years.Indications for PLT included biliary atresia (105/115),Alagille syndrome (5/115),PFIC (3/115),Caroli disease (1/115),methylmalonicacidemia (1/115) and glycogen storage disease (1/115).Most of the PLTs were living donor liver transplantation (107/115),along with OLT (5/115) and split LT (3/115).Results The diameter of the arteries was mostly less than 2 mm (98/115).Up to date,one HA thrombosis (HAT) occurred at D8 after LT and 4 cases suspected as temporal HA stenosis (HAS) around 2 weeks after LT,which manifested as low velocity (<20 cm/s) and resistance index (<0.50) by Doppler.The HAT case failed in emergent re-anastomosis,but had a spontaneous recanalization at 3 weeks and is now in good condition without biliary problem.All the HAS children recovered to normal flows at first month.All children with HA complications started warfarin upon detection,with a targeted INR between 1.5-2.0.There were 6 deaths in this series including 5 cases of infections and 1 case of graft failure.Learning curve suggested a two phases growth (first 44 cases practicing phase vs.next 71 cases mature phase),which can be attributed to experience accumulation in terms of precise of manipulation,choice of inflow arteries for better match and stronger pulsation,avoidance of length redundant,prevention of kink.All the HAT and HASs happened in practicing phase while outcomes were excellent in mature phase.Moreover,time for each anastomosis was significantly shortened in second phase from 45-70 min to 30-55 min.Conclusion Microsurgical technique is highly safe in pediatric HA reconstruction,especially for very tiny arteries.It is possible to achieve low risk of complications for a new surgeon with adequate experience in liver surgery and microsurgical training.However,more surveillance and timing anticoagulation therapy is required before the mature of microsurgical technique.

Evidence of heterozygosity and recombinant alleles in single cysts of Giardia duodenalis / Evidência de heterozigose e alelos recombinantes em cistos individualizados de Giardia duodenalis

Abstract Giardia duodenalis is divided into eight assemblages (named A to H). Isolates of assemblage A are divided into four sub-assemblages (AI, AII, AIII and AIV). While isolates of sub-assemblage AII are almost exclusively detected in human hosts, isolates of assemblage B are encountered in a multitude of animal hosts and humans. Here, we isolated single cysts of G. duodenalis from a human stool sample and found that one of them had overlaps of assemblage AII and B alleles and an unexpectedly high number of variants of the beta-giardin (Bg) and GLORF-C4 (OrfC4) alleles. In addition, one of the Bg alleles of that cyst had a fragment of sub-assemblage AII interspersed with fragments of assemblage B, thus indicating that this allele may be a recombinant between sequences A and B. Our results are unprecedented and put a check on the statement that different assemblages of G. duodenalis represent species with different host specificities.

Resumo A espécie Giardia duodenalis é dividida em oito grupos (nomeados de A a H). Isolados do grupo A são divididos em quatro subgrupos (AI, AII, AIII and AIV). Enquanto isolados do subgrupo AII são detectados quase exclusivamente em hospedeiros humanos, isolados do subgrupo B são encontrados em uma grande variedade de hospedeiros entre animais e humanos. Neste trabalho, foi constatado que, dentre diversos cistos individualizados de G. duodenalis provenientes de fezes de origem humana, um cisto continha os alelos AII e B e um número inesperado de variantes de alelos codificadores de beta giardina e GLORF-C4. Ainda, um dos alelos beta giardina desse cisto possuía fragmentos AII intercalando um fragmento B, indicando que esse alelo pode ser um recombinante entre alelos AII e B. Os resultados aqui apresentados são inéditos e colocam em dúvida o conceito atual de que os diferentes grupos de G. duodenalis representam espécies distintas com diferentes graus de especificidade por hospedeiros.

Upper limb free mini-flap transplantation for repair of finger wounds / 中国组织工程研究

BACKGROUND:Recently, the clinical repair methods of irregular wound on fingers primarily include local pedicled flap and free mini-flap of upper extremities or lower extremities. OBJECTIVE:To discuss the application of free mini-flap derived from upper limb in repairing the wound on fingers. METHODS:From December 2010 to February 2014, 12 patients with irregular wounds on 12 fingers were selected from Department of Traumatic Orthopaedics and Hand Surgery, First Affiliated Hospital of Guangxi Medical University, China. The size of wounds ranged from 1.5 cm × 2.0 cm to 3.0 cm × 4.5 cm with different degrees of bone or tendon exposure. After the debridement, 12 patients were treated by free mini-flaps of upper limb. The donor sites were directly sutured. RESULTS AND CONCLUSION:Among the 12 patients, al the wounds at recipient and donor sites were healed at stage I, and the free mini-flags survival completely. Al of patients were fol owed up for 3-6 months (average 4.5 months). Free mini-flags had good appearance and needn’t undergo secondary trimming. The active motion of fingers was improved dominantly. According to the criteria of Hand Surgery Association Society of Chinese Medical Association Society for the function evaluation of upper limb, three cases were excellent, eight were good, and only one was bad. The excellent and good rate was 91%. Free mini-flaps of upper limb are an ideal method for repair of wounds on fingers. It has no injury to normal tissue in hands, donor site is very secluded, and the short-term curative effect is good.

Clinical efficacy analysis of microsurgical clipping in treatment of ruptured intracranial aneurysm in elderly patients / 中华老年医学杂志

Objective To analyze the clinical efficacy of microsurgical clipping for the treatment of ruptured intracranial aneurysm in elderly patients.Methods 150 patients with ruptured intracranial aneurysms in our hospital from May 2009 to Apr 2013 were randomly divided into observation group and control group,with 75 cases in each group.The observation group was treated with microsurgical clipping,and the control group was treated with endovascular intervention.The efficacy,Glasgow outcome scale (GOS) and postoperative complications were compared between two groups.Results The success rate of microsurgical clipping was significantly higher in observation group than in the control group (96.0％ vs.78.7％,x2 =13.54,P＜0.05).The cure rate of GOS scale was significantly higher in observation group than in the control group (80％ vs.66.7％,x2 =4.525,P＜0.05).The incidence rate of postoperative complications was significantly lower in the observation group than in the control group (9.3％ vs.20.0％,x2 =4.58,P＜0.05).Conclusions Microsurgical clipping for the treatment of ruptured intracranial aneurysm has the high success rate and cure rate,with fewer complications and lower medical costs,and is worthy of promotion.

A rapid and reliable method for the clonal isolation of Acanthamoeba from environmental samples

Acanthamoeba are abundant in a wide range of environments, and some species are responsible for cutaneous infections, keratitis, and granulomatous amoebic encephalitis (GAE). The conventional detection and isolation of amoeba from clinical and environmental samples involves sampling and culture on non-nutrient Ágar medium. Although efficient, this system requires several transfers in order to eliminate contaminants, and is not appropriate for the isolation of individual amoeba from samples with a biodiverse community. In this study we propose an alternative method for the isolation of monocystic clones of Acanthamoeba. The propose method involves sampling, enrichment, encystment induction, and direct cysts micromanipulation and culture on Ágar plates.

Micromanipulation used in surgical procedure of transected spinal cord treated with umbilical cord blood stem cells grafts in rats / 中国矫形外科杂志

[Objective]To establish effective and reliable complete spinal cord transection models in rats,and to investigate the effects of micromanipulation used in surgical procedure of transected spinal cord treated with human umbilical cord blood stem cells(UCBSCs) grafts in rats.[Method]Human UCBSCs were obtained from umbilical cord blood of term deliveries.Sixty Sprague-Dawley(SD) female rats were divided randomly into group A(control group),group B(spinal cord transection and PBS injection),group C(spinal cord transection and UCBSCs transplantation) and group D(spinal cord transection and UCBSCs transplantation by using micromanipulation).All groups were operated,and 1 ?l UCBSCs(6?109/L ~7?109/L) were injected into the both ends of the completely transected spinal cord at the groups C and D by two different surgical procedures respectively.From 1 week to 8 week post-operation,a behavioral testing was performed weekly upon each hindlimb for all animals according to the BBB scoring system.At the 8th week,all animals were sacrificed and the spinal cords were taken out for morphological observation.[Result]No changes were observed at group A pre-and post-operation.After 2 weeks post-operation,the hindlimb motor function of groups C and D began to recover.After 3 weeks post-operation,the group D showed more improvement than group C,and the difference between groups D and C were significant in the BBB scores(P

The exploration of STR genotyping of minute cells by MiniFiler kit / 中国法医学杂志

Objective Explore the feasibility of minute cells STR typing.Methods Minute cells which were obtained by micromanipulation method were amplified with MiniFiler kit,and detected by ABI 3130 genetic analyzer.Results Ten cells can be successfully genotyped.Complete genotyping results can be obtained for one,three or five cells but with stochastic effect.Conclusion Instability is observed in minute cells genotyping,so it can not be used in actual case work.Maybe increase the quantity of DNA template can improve the success rate.

Improvement on micro-manipulation of fertilized eggs of mice / 第二军医大学学报

Objective: To improve the micro-manipulation of mouse fertilized eggs,promoting its survival rate and productivity in pseudo-pregnant mice. Methods: DNA fragments of definite concentration and size were used for micro-manipulation with conventional and improved methods, then their survival rate of manupulated eggs and productivity of pseudo-pregnant mice were analyzed and compared. Results: With improved methods, the survival rate of eggs increased from 60.3% to 79.5%, the productivity of pseudo-pregnant mice and offspring increased from 35.7% to 56.8% and 10.1% to 14.1% respectively. Conclusion: The improved methods of micro-manipulation can be used in transgenic mouse production.

Microassisted Fertilization of Human Oocytes with Intracytoplasmic Sperm Injection in In VitroFertilization Program / 대한산부인과학회잡지

In spite of much progress in in vitro fertilization and embryo transfer(IVF-ET) program,the pregnancy rate remains at 20~30%, and the endometrial implantation rate per embryotransferred at 10%. Although IVF-ET is widely applied in the treatment of coupleswith male factor infertility, it may fail in many infertile couples with normal semen parameters,and certain couples cannot be accepted for standard IVF-ET due to unfertilization orextremely low fertilization rate of oocytes. Recently, several procedures of microassistedfertilization(MAF) using micromanipulation have been introduced, and pregnancies and birthshave been obtained after intracytoplasmic sperm injection(ICSI).This clinical study was performed to develop and establish ICSI as an effective procedureof MAF in infertile couples who could not be accepted for standard IVF-ET becauseof extremely impaired semen characteristics(Group A) and because of failure in fertilizationof extremely low fertilization rate of oocytes with the conventional fertilization technique inthe previous IVF-ET cycles(Group B). From March, 1995 to December, 1996, a total of 114cycles of IVF-ET with ICSI in 65 infertile patients were included in study group, and theoutcomes of ICSI were analyzed according to fertilization rate, cumulative embryo score(CES), and pregnancy rate.In Group A, 34 patients were evaluated with semen score such as number of totalmotile sperms, and then divided into 4 groups accordingly. In 62 ICSI cycles, the numberof oocytes retrieved after controlled ovarian hyperstimulation(COH) was 12.4+/-6.8, and thenumber of oocytes optimal for ICSI procedure was 8.8+/-5.5. The fertilization rate of 65.7+/-23.6% could be obtained after ICSI. The number of embryos transferred was 4.4+/-2.2 withthe mean CES of 50.5+/-34.3 in ICSI cycles. The overall pregnancy rate was 24.2%(15/62)per cycle and 44.1%(15/34) per patient. There were no significant differences in the pregnancyrates among 4 groups. Although more mature oocytes were retrieved, the fertilizationrate was significantly lower in Group A-1 compared with Group A-IV. However, semenscore did not clearly affect the outcomes of ICSI in couples with severe male factor infertility.In Group B, the number of oocytes retrieved after COH was 10.5+/-6.1 in 49 previouscycles, and 10.8+/-5.7 in 52 ICSI cycles. In ICSI cycles, the number of oocytes optimal forICSI procedure was 8.5+/-5.1 with the fertilization rate of 72.4+/-22.5%. The number ofembryos transferred was 1.4+/-2.4 in previous cycles, and 4.7+/-1.8 with the mean CES of 50.4+/-29.9 in ICSI cycles. In ICSI cycles, the overall pregnancy rate was 30.8%(16/52) percycles and 51.6%(16/31) per patients.In conclusion, MAF of human oocytes with ICSI is a promising fertilization method forIVF-ET patients, especially with few spermatozoa for the conventional methods of in vitroinsemination and with the past history of failure in fertilization or low fertilization rate inthe previous cycles, and ICSI using micromanipulation procedures applied to human oocyteswill provide a range of novel techniques which may dramatically improve the pregnancy ratein IVF-ET program and contribute much to the effective management of infertile couples.

Impreovement in Endometrial Implantation of Humman Embryos with Assisted Hatching Using Micromanipulation in Intrauterine Embryo Transfer / 대한산부인과학회잡지

Although the fertilization rate exceeds to 80~90% with much progress in vitro fertilizaton and embyo transfer(IVF-ET) program, the prgnancy rate rmains at 20~30%, and the endometrial implantaion rate per embryo transferred at 10~15%. Recently, many attempts have been made to improve embrynic implantion after IVF-ET including serveral procedures of assisted hatching(AH) using micromanipulation, and pregnacies and births have been obtained after AH. This clinical study was performed to develop and estabilish AH as an effective procedure to improve embryonic implantioan in IVF-ET patients who had previous repeated failure of standard IVF-ET more than 2 times(Group R), were more than 37 years old(Group A), or had high basal serun FSH levels more than 15 mIU/ml(Group F). From January, 1995 to Februry, 1996, 132 cycles of AH using partial zona dissection(PZD) were performed in 104 infertile patients, and the outcomes of AH were analyzed according to pregnancy rate. The number of oocytes retrieved after controlled ovarian hyperstimulation(COH) was 9.9+/-7.1 in 71 cycles of 54 patients who had previous repeated failure more than two times(Group I: Group R,R+A,R+F, and R+A+F), 8.4+/-5.9 in 62 cycles of 46 patients whose age was more than 37 years old(Group II : Groups A, R+A, A+F, and R+A+F), and 8.7+/-6.5 in 49 cycles of 47 patients who had high basal serum FSH levels more than 15 mIU/ml(Group III:Groups F,R+F, A+F, and R+A+F). The number of embroys transferred after AH was 4.7 +/-1.8 in Group I, 4.2 +/-1.9 in Group II, and 4.2+/-2.0 in Froup III. The mean cumulative embryo score(CES) was 56.8+/-30.0 in Group I, 52.6+/-30.6 in Group II, and 52.6+/-29.9 in Group III. There were no significant differences in the numbers of oocytes rerieved and embryos transferred, and CES among 3 groups. The overall clinical pregnancy rate was 14.4%(19/132) per cycle and 18.3%(19/104) per patient. THe clinical pregnancy rate per cycle and per patient was 12.7%(9/71) and 16.7%(9/54) in Group I, 4.8% (3/62) and 6.5%(3/46) in Group II, and 26.5%(13/49) and 27.7%(13/47) in Group III, and there was a significant difference between Group II and Group III. In conclusion, AH of human embryos using micromanipulation might be promising for IVF-ET patients, especially with the past history of repeated failure, old age, and high basal serum FSH level and AH will provide a range of novel techiques which may dramatically improve the implanatation and pregnancy rates in IVF-ET program and contribute much to effective management of infertile couples.